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The answer to this very much depends on what you are charging for. To find an answer, you need to identify (a) the net costs you wish to recover: e.g. capital costs (to be recovered over what period?, at what rate of interest?), running costs, salaries of direct (em technician) and indirect (clerical, admin.) support staff including overheads, any charges for space occupancy and services etc., less any subsidies (b) make a realistic estimate of the maximum number of hours per year that a charge can be recovered from. The answer is a/b. Simple, isn't it? Well, no. In practise you will probably find that b is dependent on a/b since all categories of users are sensitive to price, but you will have to model this differently for different categories of users.
Our charges per hour for self-help access to a Philips CM120 Biotwin are as follows: Departmental £15, Other departments £21, other publicly-funded research organisations £35, industrial £70. Non-attenders incur full charge for the period booked. These numbers were not exactly arrived at using rocket science. We charge extra for all materials consumed (film, processing) and for direct user-support from a technician. We do not include capital costs recovery in our charges.
I would strongly advise imposing a special concessionary charge on users who insist on tipping their specimens and retaining circlips into the guts of the Compustage! An electronically-operated trap door beneath the operator's seat is also an attractive option.
Chris
} From: "Ping Li" {pli-at-is.dal.ca} To: {Microscopy-at-sparc5.microscopy.com}
Folks: A Happy New Year to you all I thought I should let you all know about the Third annual course in Quantitative Fluorescence Microscopy to be taught between june 17 and 22th 2001 at the Mount Desert Island Marine Biology Laboratories in Arcadia National Park in Maine. This team taught course led by Dave Piston (Vanderbilt), Mike Nathanson (Yale), Guillermo Romero (Pittsburgh) and myself focusses specifically on the development and application of modern fluorescence microscopic methods. This six day intensive course covers all aspects of the technology from microscope and dye design, cameras, confocal microscopy, live cell microscopy, multiphoton microscopy and GFP. Considerable attention is also given to quantitative analysis in 2 and 3 dimensions and time. The specific focus of the course allows an in depth treatment of these methods. The goal of the course it to teach students how to best implement these methods within their labs, using either their own cells and tissues or using material supplied by the course. An extensive array instrumentation, provided by all the major microscope and associated software, hardware and camera manufacturers will be available for students to use. For the last two years it has been a very successful event and so we have decided to give the course again. A full description of the course lectures together with lecture outlines, registration etc. is available at http://sbic6.sbic.pitt.edu/microscopy/ I would encourage you to spread the word, or sign up if you are interested. The total number of students is limited to 25, enrollment is decided by the course faculty. If you have any further questions please feel free to contact me Thanks Simon
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 http://sbic6.sbic.pitt.edu -----------------------------------
} There is a position for an EM technician at the Electron } Microscopy Core lab of the Biotechnology Program at the University of } Florida in Gainesville. The laboratory is mostly a fee-for-service lab } serving the needs of biological, biomedical and agricultural scientists } at the university. Applicants must be skilled in the preparation of } biological samples for both scanning and transmission electron } microscopy. Experience with both PC and Mac as well as website } management is desirable. Experience with digital imaging and } fluorescence microscopy also a plus. } This is a full time, permanent position with standard benefits of } State of Florida employees. } } For further details, reply to this message or contact Greg Erdos } at the number below. } } }
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
As part of the planned makeover of our building, it has been proposed that the various and somewhat distributed microscopy facilities in our building - SEM, TEM, Confocal, fluorescence, luminescence imaging, darkroom etc. etc. might be brought together to form a biological imaging facility, thereby benefitting from some improvements in supervision, security and user support. Part of that proposal was to set up a computing lab with fileserver and work- stations networked to the instruments and dedicated to image storage and off-line image and spectral processing and analysis. However, there is great pressure on space in this building, and it has been suggested to me that computing for imaging and spectroscopy does not need to be separated from general purpose computing these days. What are your opinions about this? Pro or con?
Happy New Millennium Chris ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
Hi Tim, I like your idea of a separate step for the UA*. Though 1% may be overkill. We routinely use a cocktail for freeze-sub consisting of: 1) 4% osmium tetroxide in HPLC grade acetone 2) 0.1% uranyl acetate also made with HPLC grade acetone - make this up the day before the freezing so it has time to go into solution before chilling. Mix together 1:1 for wonderful contrast. *We have to use Radiation Safety Services for disposal of this cocktail because our hazardous waste folks refuse to deal with the combination of OsO4 and UA.
Another angle - You didn't mention the type of resin you're using. Freeze-sub tissue is often murky looking (low contrast) when it is embedded in 100% Spurr's. We use Araldite/Embed 812 (an EMS kit) or a mixture of equal parts Spurr's and Araldite/Embed 812.
hope this helps, Beth
} Hello Out There: } } I am currently involved with a cryo substitution project with human red } blood cells. I have replaced the non crystalline ice with 2% Osmium in 100% } acetone and have OK morphology, but the contrast is a bit low. I am } thinking about trying to follow the Osmium step with uranyl acetate, before } embedding in plastic. Does any one out there know if 1%uranly acetate will } be OK in 100% acetone? Thanks, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I use Ralph type glass knives for plastic sectioning of biological samples. I am experiencing problems in making a proper knife. I have a TAAB histoknifemaker (also says Reichart-Jung). Is there anyone willing to offer expertise in this area?
Within the last couple of days (and, therefore, not in the recently posted archives) someone posted a note about a used chiller that they had for sale. I've lost the note. If still available, could you please repeat the note to my email address: edsworth-at-aol.com Thanks.
Ed Holdsworth General Mgr. SEMTEC Laboratories, Inc.
Those of you who need to process electron diffraction ring patterns (recorded from thin polycrystalline or amorphous samples in a TEM) might be interested in the FREE program (for IBM PC, Windows op. system) that can be downloaded from my home page (below). The program produces XRD-like output from digitized SAED ring-patterns and compares to markers showing the positions and intensities of known phases. The newest version (1.2.0) of this ProcessDiffraction program provides easier operation and new functionality:
* Built-in hints: what to do next (and how). Can be switched off.
* File-name and path can be specified for output. Default paths are suggested separately for input SAED, input Marker and output. Manually selected paths can be stored as default (during exit).
* No limit on BMP file-size.
* The automatic peak-list can be manually extended using Cursor.
* Selection of individual points (in the SAED for reading-out of d-value and establishing correlation between distribution and SAED) is aided with a Cursor with increased contrast and magnification.
* Units of X-axis can be changed to the physically meaningful scattering vector [1/A] (in contrast to pixels).
* Several net distributions can be compared
* Individual markers have different colors.
* Renormalized Gross intensity can be calculated by excluding pixels with zero intensity (i.e. corresponding to beam-blocking pin) from the averaging process.
* Renormalized Net intensity can be calculated. Here points with intensity above the average (over a given circle) are only included in the averaging. Average intensity of bright pixels within a discontinous ring are only calculated this way, giving more realistic intensity of a diffraction line for comparison purposes.
* Show menu is included to select which curves to show in the graph.
* Possibility to cancel is introduced at several operations.
* Automatic (fast) refinement of centre is included. In dubious cases manual selection of peak and valley is asked for.
* Calculation is double precision (basis of later extension of the software for input files with several bytes/pixel).
* Several error conditions are eliminated (crop, conflicting user requests, ..)
I hope you will like the modifications and find this program useful and easy to use. Download FREE as before from the same site.
I would appreciate having your comments and/or suggestions. Best regards:
Dr. Janos L. Labar Research Institute for Technical Physics and Materials Science H-1121 Budapest, Konkoly-Thege u. 29-33 Postal address: H-1525 Budapest-114, Po Box 49 Tel: (36)(1) 392-26-92 Fax: (36)(1) 275-49-96 Fax/phone: (36)(1) 395-92-32 home page: www.mfa.kfki.hu/~labar
University of Connecticut Institute for Materials Science
Postdoctoral Position in Electron Microscopy
The Institute for Materials Science (IMS) at UConn is an interdisciplinary center with the threefold mission of fostering education, research and outreach in all areas of the materials sciences. The Institute has a vacancy in the EM laboratory for postdoctoral researcher to work on a DARPA-funded program on Ni-based superalloys and a variety of smaller industrial projects. Candidates should hold a PhD in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of electron microscopy techniques. A candidate with experience in the microstructural analysis of Ni-based superalloys using SEM and TEM would be preferred. The appointment is for one year in the first instance and is available immediately. Screening of the applications will begin immediately and will continue until the post is filled. Applications from under-represented groups, including minorities, women and people with disabilities are encouraged.
Interested candidates should send a curriculum vitae, including publication list, and the names of at least three referees with postal addresses, telephone numbers and Email addresses to: Prof. M. Aindow, Institute for Materials Science, University of Connecticut, 97 North Eagleville Road, U-3136, Storrs, CT 06269-3136 USA. Email: m.aindow-at-uconn.edu
We are a clinical diagnostic TEM laboratory serving a provincial network of hospitals. We would like to move away from conventional darkroom printing of images and wish to convert negatives to digital files. We are looking for a negative scanner that will:
1. Accept 3 X 4 1/4 inch sheet film negatives (Kodak 4489 film).
2. Comes with software support that is adaptable to a Windows format.
3. Must be priced under $3000.
4. Must provide the best quality resolution for diagnostic results (i.e. 3200 X 3600 pixel; wide linear dynamic range).
I look forward to any information and advice you can provide.
My problem is a poor knife edge. The knife looks fine under casual observation, or with small magnification, but when it comes time to actually do some sectioning, I get a lot of chattering, or streaking, or both. I know make as many test blocks as I do sample blocks, so I don't waste samples with all the bad glass knifes. I've gone through several boxes of glass over the past two months, and only get a useable knife every 15 to 20 breaks. The same knife breaker has worked fine in the past, and I have changed the scoring wheel, and checked all that I can.
This knife breaker is also very hard to get parts for. The only obvious problem is that one of the pressure points has a rubber pad that is unevenly worn. I don't see how that would cause my problem. Again, the knifes look good to the naked eye.
The glass I use is from Electron Microscopy Sciences. They are called ultra glass knife strips 6.5mm x 25mm x 400 mm.
I guess I'm just looking for someone who has had a similar problem, and fixed it in some manner. Could I have gotten a bad batch of glass? I don't think it is very likely, I've used several boxes, but they may all be from the same batch, for all I know. Is there another brand of glass, or breaker that others have found produce reliable knives?
Thanks for any and all help
-Patrick Brownell } ---------- } From: Katja M. Wolski[SMTP:kwolski-at-hsc.usf.edu] } Reply To: kwolski-at-hsc.usf.edu } Sent: Wednesday, January 03, 2001 4:26 AM } To: Brownell, Patrick } Subject: Re: Ralph Glass Knife problems } } I use a different knife machine, but what's the problem you're having? } Are you } not getting a good knife edge or what? } } Katja } } } } "Brownell, Patrick" wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi all! } } } } I use Ralph type glass knives for plastic sectioning of biological } samples. } } I am experiencing problems in making a proper knife. I have a TAAB } } histoknifemaker (also says Reichart-Jung). Is there anyone willing to } offer } } expertise in this area? } } } } -Patrick Brownell }
I recently acquired a Zeiss OPMI with floor stand. I inspected the bulb superficially before turning it on and it seemed OK, and I confirmed the voltage was in range beforehand. However, as soon as I applied power, the filament left the prongs and the bulb was dead.
It was a Zeiss 390158 bulb, a 6v 30w triangular bayonet. It's quite possible the bulb was mechanically shocked before I got it. Was this a not unusual failure, or is there some kind of warm-up procedure that might've saved it?
A hunt through my junk boxes fortunately turned up a replacement, and it worked straight-away, even though it looks like one of the spring-like wires worked its way off the filament prongs, where the filament meets the prong wire.
For the day that this one dies, though, I'd also like recommendations for a place to purchase a replacement.
Greetings. Can anyone suggest a good all around sputter coater for general SEM use? I usually use 50 Angstrom Pt, or Carbon for EDS. Thanks in advance.
Tracy E. Anderson Microscopy Specialist Surface Characterization Dept. SurModics, Inc. www.surmodics.com tanderson-at-surmodics.com
Anyone else happen to watch "Mysterious Ways" earlier this week. One character was chatting with another about the fantastic new TEM they were using with the "Kevex EDS system with the twin Quantum 10-mm2 detectors". I had to listen quick to hear it.
Granted, its a bit dated (Thermo-Noran is now the name and they don't list the Quantum on their web page), and some of their other references to other techniques were of questionable accuracy. Still, I was tickled to see a reference to EM on TV. I remember a few references on "Quincy", but not a lot since.
Disclaimer - I don't have a twin Quantum system, but am still using a 10-mm2 Quantum on our JEOL-840.
Hi all: Can anyone tell me who the current manufactures of ultra microtomes are? -- Regards, Gregory Rudomen Technical Specialist Electron Microscopy State University of New York at Stony Brook University Microscopy Imaging Center Stony Brook, NY 11794-8088 631-444-7372 Greg-at-umic.sunysb.edu ************************************************* Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. *************************************************
Bulbs of the most exotic sort can be found at: 1. Bulbtronics;(516)-249-2272 2. Bulb Direct; 1-800-772-5267, or info-at-bulbdrect.com
Good luck,
Sam Purdy Technical Center, National Steel Corp. Trenton MI 48183 Voice; 734-676-2682 FAX 734-676-2030 E-mail spurdy-at-nationalsteel.com } ---------- } From: John Foust } Sent: Wednesday, January 2001, 11:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Zeiss lamp failure question } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I recently acquired a Zeiss OPMI with floor stand. } I inspected the bulb superficially before turning } it on and it seemed OK, and I confirmed the voltage } was in range beforehand. However, as soon as I } applied power, the filament left the prongs and } the bulb was dead. } } It was a Zeiss 390158 bulb, a 6v 30w triangular } bayonet. It's quite possible the bulb was } mechanically shocked before I got it. Was this } a not unusual failure, or is there some kind of } warm-up procedure that might've saved it? } } A hunt through my junk boxes fortunately turned up } a replacement, and it worked straight-away, even though } it looks like one of the spring-like wires worked its } way off the filament prongs, where the filament meets } the prong wire. } } For the day that this one dies, though, I'd also like } recommendations for a place to purchase a replacement. } } - John } }
Tracey E. Anderson wrote: ====================================================== Greetings. Can anyone suggest a good all around sputter coater for general SEM use? I usually use 50 Angstrom Pt, or Carbon for EDS. Thanks in advance. ====================================================== SPI Supplies manufactures a modular coater that is robust, requires little maintenance, and will do all of the precious group metals, including Pt. Carbon is done via the carbon coater module. A number of other firms manufacture sputter coaters for SEM applications and they can all be found on the following two vendor data bases:
Microscopy Vendor's Data Base http://207.137.96.185/mvd/vendors.html
MicroWorld Resources and News http://www.mwrn.com/
Disclaimer: SPI Supplies manufactures sputter and carbon coaters for SEM laboratory applications.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
MAS membership renewal application forms were mailed late last week. Enclosed with your application is the 2000 Membership Directory. Please make any corrections on your form and return it along with your dues in the envelope provided. My sincere apology for the late mailing.
If you have any questions, do not receive your renewal packet, or are not a member of MAS and would like to join, please contact me either at work or at:
MAS Membership Services PMB 141 2101 W. Broadway Columbia, MO 65203-1261 1 (800) 4MASMEM masmembership-at-excite.com
Thanks, Lou Ross Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.missouri.edu/~geosclmr/ebaf.html
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: John Foust {jfoust-at-threedee.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
What constitutes a "good all around" sputter coater is perhaps somewhat vague? If you are looking for something that is easy to use and provides consistent results, that may be a problem.
New units are in the $4K to $8K price range. This depends on whether it has a pump included, thickness measuring unit (not all that usefull, IMO) and type of target, and dual mode or single plating mode. These units are sputter coaters, not evaporation units.
I searched for what it sounds like you are doing the same. I narrowed the search to a Denton Desk II or an Anatech Hummer VII. I settled on a used Hummer. This is an obsoleted unit. But after some initial hickups, this coater works really well. Everything is contained in one unit. It has old metal gate CMOS digital ICs (still available) and an Edwards or Alcatel small pump inside. It will do etching and plating. What I like about this unit is that it will do plating at a specified rate with a specified end point thickness. These settings must be validated to be absolutely correct. The rate and power can be adjusted via pots to hone the correlation of settings and results.
Newer units cost tons of money and do not do a proportionately better job. They are basically manual units. Set the time, and make the run. No idea of thickness. Pump is optional...right. Thickness measurement options are a waste of money in my opinion. The results are not repeatable. And the most common measuring method is a quartz crystal. This is a consumable item.
You have a tough decision, based on my experience. I would suggest that you look for a used Hummer VII or a Desk II. I just don't think that the newer units are all that great of a value. Nor are they all that user friendly. But YMMV.
gary g.
At 12:44 PM 1/3/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am participating in an art project that incorporates micro-organisms (e.g. euglena although larger ones e.g. hydra are feasible) as actors. The video from the microscopes/cameras will be streamed in realtime via the Internet.
Image quality is important to us, cost less so although there shall be several stations in different geographical locations.
Should one wish to suggest a particular microscope and camera we would be delighted. (The software isn't an issue)
A European company -Digital Biomedical Imaging Systems AG (DITABIS), produce a plate scanning system that may suit your future needs, but I'm not certain whether you can scan negatives with it. It's an imaging system to replace using negatives; I don't know if they have a distributor State-side but a while back I made contact with a British company at the Micro 2000 conference, UK, who are distributors for the system in the UK and they were willing to let customers have a trial run with the scanning plates.
A web site http://www.ditabis.de/first.html explains the system
The UK company is
ISS group services Pellowe House Francis Rd Withington Manchester M20 9XP
Phone:+44 161 445 5442 Fax:+4 161 445 4914
They don't appear to have a web site
Hope this info is of some use + I stress that I have absolutely no commercial or other interests with the above companies.
Mr. Cameron Hind Research Scientist Advanced Technologies group Baxter R & D Europe S.C.R.L. Rue du Progrès 7 1400 Nivelles Belgium
Hi John In reference to your request for replacement bulb 39-01-58...I currently use Bulb Direct for all my lighting needs. 1-800-772-5267 WWW.BULBDIRECT.COM This company is very good on delivery (2 days) and have the best prices I have run across.
I have no affiliation with Bulb Direct just satisfied customer.
Can anyone advise me on what chemical sample I can use to test the (in)accuracy of EELS quantification for oxygen. Is there an oxide for which there is abselutely no doubt on the oxygen concentration. Preferably with a metal which has a good EELS peak (below 1 keV). The goal is to have a reference with a reliable metal/O ratio.
Facility Manager Position Electron Probe Instrumentation Center Northwestern University
The Electron Probe Instrumentation Center (EPIC) at Northwestern University has an immediate opening for a full-time facility manager to assume the responsibility of managing the advanced EM laboratory and facilitate/initiate advanced TEM research. Responsibilities also include supervision of two technical staff in SEM and specimen preparation. The EPIC facility serves over 100 users in all aspects of Scanning and Transmission electron microscopy. The role of the Facility Manager is to provide leadership in advanced electron microscopy in materials research, and assist users with their advanced TEM needs, including the following: teaching/assistance in: HRTEM, nanodiffraction, EDS, EELS, image processing, etc. A strong laboratory development component, research initiatives and commitment to teaching are associated with this position. All EMs in EPIC are under full service contract. Thus, the duties include mainly training students/users in their research needs, development of specialized techniques and applications for materials research, and minor records and fiscal management. A PhD in physical sciences/engineering is required. The candidate must have hands-on experience in all aspects of advanced TEM as well as digital acquisition, processing and computer assisted techniques. All levels of experience will be considered. Compensation commensurate with experience and qualifications. Promotion to research faculty position is possible with proven research credentials. Send cover letter, resume and three references to: Prof. Vinayak P. Dravid, Director EPIC Materials Science & Engineering Northwestern University, 3013A MLSB Evanston, IL 60208 E-mail: v-dravid-at-northwestern.edu Fax: (847) 491-7820 http://epic.ms.nwu.edu/epic/index.htm
Northwestern University is an Affirmative Action/Equal Opportunity Employer. Hiring is contingent upon eligibility to work in the United States.
I am a research assistant at Catholic University and have been attempting for the past few months to thin section alteration layers, (in cross section,) of simulated nuclear waste glasses subjected to vapor hydration testing. These altered layers are typically comprised of multiple phases, frequently composed of various alumino-silicates, and are brittle and porous. They range from a few microns to hundreds of microns. When fragmented off the test coupon, usually a few microns (~1-20um) of glass remain adhered to altered region. We are interested in this transition region between the glass and altered layers.
Dozens of attempts have been made to section these materials once embedded. Variations of impregnation protocols have been tried with recipe variations utilizing Spurr’s resin. Block faces on average have been ~200x500um with particles sizes anywhere from a few microns to ~200um. Utilizing a Sorvel MT-2, cutting speeds between 0.25 and 2.5mm/second have been attempted, (the full range of the instrument.) The diamond knife is new. The advance has been altered between 20nm and 250nm – producing the appropriate interference colors, but with voided regions wherever the particles are situated. I have tried everything I could think of to facilitate infiltration/adhesion of these particles within the embedment medium. After thousands of sections from 80+ blocks and several protocols, I haven’t had the slightest glimmer of success. I know the very thing we have been attempting has been successfully done at ANL. So, I implore anybody that has any specific insight, simple thoughts, or longshots to let me know.
Sincerely,
Cavin T. F. Mooers Research Assistant The Catholic University of America 434 Hannan Hall Washington, D.C. 20064 (202) 319-5346 phn (202) 319-4469 fax
And now, from the Last Hope Department, I was wondering if anyone out there has an old Leitz Tas Plus Image Analyzer? This would be of mid-'80's vintage, comprised of a bench with a pop-up monitor with light pen, a nice microscope with built-in video capability, and a big black electronics box about four feet tall (containing 2 8-inch floppy drives - like I said, this thing is old). Long story short, ours is broken, and we can't even begin to fix it without schematics (some clever soul -no, not me - tossed them out years ago). If anyone happens to have a set of the electronics schematics for one of these things hanging around, please contact me ....we'll talk....
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) P.O. Box 1006 Dartmouth, Nova Scotia Canada B2Y 4A2
Gatan, Inc., the technology leader in digital imaging, analytical spectroscopy and icon beam milling applications for electron microscopy in Pleasanton, CA., is seeking candidates for a TEM Laboratory Manager. This person will manage the TEM applications laboratory. Responsibilities include coordinating all activities related to customer demonstrations and training, Applications material generation and supporting the R&D department utilization of the TEM systems. Some travel within and outside the USA is required. Applicants should have significant TEM and CCD imaging experience. Biological, materials or semiconductor experience would be useful. Familiarity with existing vendors, consultants, competitors, etc in the industry is a plus. Knowledge of Gatan's software and hardware applications. A proven track record in both planning and managing a laboratory environment and outstanding computer/PC skills. Must be comfortable with current development environments and development tools to work intelligently with engineering. Must have excellent overall PC computing skills, including a thorough familiarity with market tools in document composition, database design and presentation management. PhD, desired; technical background preferred. Salary: Base salary plus bonus commensurate with experience.
Interested candidates should send, email or fax their resume to:
GATAN, INC Attn: HR Department 5933 Coronado Lane Pleasanton, CA. 94588 Fax: (925) 463-0204 Email: hr-at-gatan.com www.gatan.com Carlotta Daniels GATAN, Inc. Human Resources
Calvin, Hard materials such as glass can be readily microtomed with a diamond knife (even diamond coated silicon in Microscopy Research and Technique, vol. 31, p. 308, 1995). Any good diamond knife will work with meticulous and careful technique, but experience has shown that 35 degree knives yield the best results with hard and ultra-hard materials.
It sounds as though your samples may be large and adhesion to the epoxy is poor. In order to microtome non-porous materials such as glass, you should first minimize the cross-sectional area to be sectioned. An easy way is to do this is to pop concoidal micro-chips from the surface. These tend to be very thin at the edges and can be further broken to form pointed thin samples. Another critical element is to improve adhesion of the sample to the resin (Spurrs) with the use of an adhesion promoter such as Dow Corning Z-6040. Submerging the sample in a 2% solution of Z-6040 (50/50 ethanol and H2O) for 30 minutes allows a surface reaction that promotes adhesion of glass to epoxy. Remove samples from the solution, air dry, and embed in Spurrs. Then, microtome using standard procedures with sectioning speed reduced to 0.1 mm/s.
Cheers,
Phil Swab, Sr. Engineer Engineering Development Group Deposition Sciences, Inc 386 Tesconi Court Santa Rosa, CA 95401
-----Original Message----- } From: Cavin Mooers [SMTP:cavinm-at-vsl.cua.edu] Sent: Thursday, January 04, 2001 6:30 AM To: Microscopy-at-sparc5.microscopy.com
Dear Microscopists,
I am a research assistant at Catholic University and have been attempting for the past few months to thin section alteration layers, (in cross section,) of simulated nuclear waste glasses subjected to vapor hydration testing. These altered layers are typically comprised of multiple phases, frequently composed of various alumino-silicates, and are brittle and porous. They range from a few microns to hundreds of microns. When fragmented off the test coupon, usually a few microns (~1-20um) of glass remain adhered to altered region. We are interested in this transition region between the glass and altered layers.
Dozens of attempts have been made to section these materials once embedded. Variations of impregnation protocols have been tried with recipe variations utilizing Spurr's resin. Block faces on average have been ~200x500um with particles sizes anywhere from a few microns to ~200um. Utilizing a Sorvel MT-2, cutting speeds between 0.25 and 2.5mm/second have been attempted, (the full range of the instrument.) The diamond knife is new. The advance has been altered between 20nm and 250nm - producing the appropriate interference colors, but with voided regions wherever the particles are situated. I have tried everything I could think of to facilitate infiltration/adhesion of these particles within the embedment medium. After thousands of sections from 80+ blocks and several protocols, I haven't had the slightest glimmer of success. I know the very thing we have been attempting has been successfully done at ANL. So, I implore anybody that has any specific insight, simple thoughts, or longshots to let me know.
Sincerely,
Cavin T. F. Mooers Research Assistant The Catholic University of America 434 Hannan Hall Washington, D.C. 20064 (202) 319-5346 phn (202) 319-4469 fax
Hi, we have some imaging problems with Zeiss LSM 410. Sometimes, when we collect an image series, one or more of the images had different sized lighter or darker bands across the images. This happened with both green and red channels (488 nm and 543 nm). It occurred randomly. In addition, sometimes, the image suddenly totally whited out during the scanning. In which case, the up part of the image looks normal, but the lower part is white. In all cases, there were no indication of any changes in pin hole size, contrast, brightness, and laser power based on the readings showed in the LSM program. I am not sure what is wrong. Could be the scanner? video card? or may be something else? Could any of you please help me and tell me what do you think the problems are? Any information is highly appreciated.
I am using regular EPSON Expression 1600 flatbed scanner with "transparency adapter". It has 1600 optical dpi resolution and pretty fast. To eliminate the Newton rings, you may use special cassettes to hold the film. Unfortunately, EPSON does not provide the cassettes for standard EM film. You have to made them in machine shop (not big deal, actually). I do use some EPSON cassette, which is bigger than my film in one dimension, but it works well. This scanner comes with very good software and you may scan your images directly into Photoshop or any other suitable program. I am scanning in "positive transparency", 12-bit (Hi-Fi) gray, 1600 dpi mode and than adjust the levels in the Photoshop (to remove the "empty" levels of gray), transfer in 8-bit mode and save. Usually, I prefer to keep the original 12-bit image as well, but it's huge - about 50 Mb each. I am using 2.6 or 5.2 Gb MO disks for storage.
I hope it will help.
Sergey
P.S. I have no interest in EPSON products, just happy user.
At 11:24 AM 1/3/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
This equipment specially designed to work with "image plates". Nothing related to the negatives. Moreover, they claimed that their system is supposed to substitute the regular photo-process (and in some instances that works better, linearity and sensitivity, for instance). Only one disadvantage - very pricey and time consuming.
Sergey.
At 09:50 AM 1/4/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I think Phil is right. You better collect really a small smaple which is like a needdle. At the frond end of your needle, you will have the altered layer plus a little glass. Crack your corroded glass samples you can always find out a tiny needle suitable for ultramicrotomy. You may read our paper in Journal of Nuclear Materials (Vol. 254, pp. 249-265, 1998) for your reference.
Good luck,
Weiliang Gong Center for Radioactive Waste Management University of New Mexico 1001 University Blvd. Se Albuquerque, NM 87106
Hello- Does anyone out there know the scintillator sizes for the upper and lower secondary electron detectors on a Hitachi S-4500? Also, does anyone have an opinion on whether the single crystal YAP (or YAG) scintillators are worth 10x the money of a P-47? The catalogues say they last longer and have better S/N even though they produce less signal than the P-47. If I switched, would there be adjustments I would need to make to my SEM? Thanks for your help.
We are considering purchasing a Nikon Coolpix digital camera for use on light microscopes. We would like to know if anyone has experience using these cameras for fluorescent microscopy. Is the meter accurate at low light levels? Thanks.
Richard Demaree, Ph.D. Dept. Biological Sciences Calif. State Univ., Chico Chico, CA 95929-0515 530-898-5812 rdemaree-at-csuchico.edu
I am trying to find a way to perform immunofluorescence in plant protoplasts for both cytosolic and peroxisomal proteins.
My problems are as follows:
1) Fixing of the protoplasts prior to immunofluorescence is difficult; fixation in 4% paraformaldehyde, 350 mM mannitol, and 50 mM sodium phosphate, pH 7 for 50 min followed by treatment with 0.1% triton X-100 for 10 minutes yields protoplasts that appear to be aggragated and broken.
2) Fixing of protoplasts expressing cytosolic GFP, as detailed above, loose their GFP-fluorescence.
Does anyone have an established method for plant protoplast immunofluorescence?
Thanks
Kristi Snell
-- Dr. Kristi D. Snell Metabolix, Inc. Cambridge, MA
Greetings. I would like to look at a cross section of a standard sized glass slide under TEM. Does anyone know how I could prepare such a sample? Will a microtome work? Any suggestions would be very much appreciated.
Tracy E. Anderson Surface Characterization Specialist SurModics, Inc. 952.829.2720 Voice 952.829.2743 Fax tanderson-at-surmodics.com
There are a number of standard methods that will work.
Microtomy should work. There has been a recent thread about some porous glass. You should see the postings by Phil Swab. He also has a Journal article that you might be able to get a copy from Stacie Kirsch on how to do cross section of glass using microtomy. He recommends a 35 degree knife. Basically, you create small shards from the surface of the glass, mount them and then cut them. If you know how to microtme and do not know any of the other standard cross sectional preparation techniques, I would go with this one.
You could use the small angle cleavage technique. It is relatively inexpensive and easy to learn. It also works well with glass.
You can use the standard cross sectional method of Bravman and Sinclair. You can take two pieces of your glass slides about 5-10 mm x 10-20 mm and epoxy them together. I use Epo-Tek 353ND, but you can also use N-bond 610. I prefer to use a stack of silicon on one side for a number of reasons: 1) I believe that the Si helps reduce the charging effects, 2) I believe that the Si helps prevent the glass from heating under the beam, 3) I can use the Si to gauge the thickness of the sample during dimple grinding, and 4) the Si can help me find the orientation of the sample relative to the beam that puts the surface parallel to the beam. Cut the samples with a thickness of about 400-500 um with a diamond wheel cutoff saw, hand lap them with a hand grinding tool with parallel sides to between 60-90 um thick, dimple grind them to about 5 um (the Si will show a rich red color in transmitted light), and ion mill to perforation and electron transparency.
You can use Tripod polishing with a low angle ion mill clean-up. This takes some learning to do properly and specialized tools.
You can use an FIB. Glass cuts reasonable well without too much charging affects. Protect the surface with a relatively thick sputter coat of gold or Au-Pd before you put it in the FIB, especially if you use a single beam FIB.
With your finished sample, I found that at 125 kV I had to put a light carbon coat on the sample to prevent softening of the glass under the beam. AT higher voltages (200 kV or better), you don't need to coat the sample for charging or for eliminating the heating effects.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Tracy Anderson [mailto:tanderson-at-surmodics.com] Sent: Thursday, January 04, 2001 4:29 PM To: 'Microscopy ListServer' Subject: TEM sample prep for glass slide
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
Greetings. I would like to look at a cross section of a standard sized glass slide under TEM. Does anyone know how I could prepare such a sample? Will a microtome work? Any suggestions would be very much appreciated.
Tracy E. Anderson Surface Characterization Specialist SurModics, Inc. 952.829.2720 Voice 952.829.2743 Fax tanderson-at-surmodics.com
} This equipment specially designed to work with "image plates". Nothing } related to the negatives. Moreover, they claimed that their system is } supposed to substitute the regular photo-process (and in some instances } that works better, linearity and sensitivity, for instance). Only one } disadvantage - very pricey and time consuming.
Yes, they are better for diffraction work (high dynamic range, linearity). But why time consuming? You just put the plates in the scanner and go do some other things. When you are back you have all the images scanned. No development and stuff like with films.
Best regards,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp ---------------------------------------------------------------------
} I am using regular EPSON Expression 1600 flatbed scanner with "transparency } adapter". It has 1600 optical dpi resolution and pretty fast.
Have you tested the MTF of the EPSON? I'm interested what are the actual resolutions X,Y.
Best regards,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp ---------------------------------------------------------------------
We have a Tektronix Phaser 780 color laser-printer which is currently out of service. We can not find anyone in Oklahoma who can service it; Xerox, who purchased the Tektronix printer line, is not providing service for this printer.
I wonder if any of you who have this printer have a service source? We seek a service center in the Oklahoma-Texas-Kansas-Arkansas-Misouri area, but are nearly desperate enough to ship this thing anywhere in the U.S.
Thanks, in advance, for any help you provide.
Winton Cornell
--
Winton C. Cornell, Ph.D. Department of Geosciences College of Engineering and Natural Sciences The University of Tulsa Tulsa, OK 74104 phone: 918-631-3248 fax: 918-631-2091 e-mail: winton-cornell-at-utulsa.edu
Sorry I do not know the sizes of the scintillators in the 4500/4700 SEM but the only change you should see is that you run the "contrast" at a lower level than you did prior to the change.
Work we have carried out suggests an improvement compared with the conventional product. Scintilltors start when new with a high signal that falls off quite quickly to start with then there is a gradual decline over the life of the scintillator. In my mind people do not change, or in your future case clean, the scintilltors enough.
Good luck
Steve Chapman Senior Consultant Protrain For consultancy and training by professionals World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
In an episode of the "X-Files" the EM operator looks up from the TEM and shows Scully the alien virus. It turns out to be an SEM of a pollen grain.
After telling the story to visitors I remind them that any scientist who reveals important information to either Mulder or Scully must take great care when driving home!
Dave
On Wed, 03 Jan 2001 14:25:03 -0600 Warren E Straszheim {wesaia-at-iastate.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Anyone else happen to watch "Mysterious Ways" earlier this week. One } character was chatting with another about the fantastic new TEM they were } using with the "Kevex EDS system with the twin Quantum 10-mm2 detectors". I } had to listen quick to hear it. } } Granted, its a bit dated (Thermo-Noran is now the name and they don't list } the Quantum on their web page), and some of their other references to other } techniques were of questionable accuracy. Still, I was tickled to see a } reference to EM on TV. I remember a few references on "Quincy", but not a } lot since. } } Disclaimer - I don't have a twin Quantum system, but am still using a } 10-mm2 Quantum on our JEOL-840. } } Warren S. } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Perhaps you could get a lab with no problems to make some knives with your glass and then test them in their lab and yours (get them to send some of their glass to you as well to test your knifemaker). That should show if it is the glass, the knifemaker or the ultramicrotome that is at fault.
Dave
On Wed, 3 Jan 2001 08:04:10 -0800 "Brownell, Patrick" {patrick.brownell-at-weyerhaeuser.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My problem is a poor knife edge. The knife looks fine under casual } observation, or with small magnification, but when it comes time to actually } do some sectioning, I get a lot of chattering, or streaking, or both. I } know make as many test blocks as I do sample blocks, so I don't waste } samples with all the bad glass knifes. I've gone through several boxes of } glass over the past two months, and only get a useable knife every 15 to 20 } breaks. The same knife breaker has worked fine in the past, and I have } changed the scoring wheel, and checked all that I can. } } This knife breaker is also very hard to get parts for. The only obvious } problem is that one of the pressure points has a rubber pad that is unevenly } worn. I don't see how that would cause my problem. Again, the knifes look } good to the naked eye. } } The glass I use is from Electron Microscopy Sciences. They are called ultra } glass knife strips 6.5mm x 25mm x 400 mm. } } I guess I'm just looking for someone who has had a similar problem, and } fixed it in some manner. Could I have gotten a bad batch of glass? I don't } think it is very likely, I've used several boxes, but they may all be from } the same batch, for all I know. Is there another brand of glass, or breaker } that others have found produce reliable knives? } } Thanks for any and all help } } -Patrick Brownell } } ---------- } } From: Katja M. Wolski[SMTP:kwolski-at-hsc.usf.edu] } } Reply To: kwolski-at-hsc.usf.edu } } Sent: Wednesday, January 03, 2001 4:26 AM } } To: Brownell, Patrick } } Subject: Re: Ralph Glass Knife problems } } } } I use a different knife machine, but what's the problem you're having? } } Are you } } not getting a good knife edge or what? } } } } Katja } } } } } } } } "Brownell, Patrick" wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Hi all! } } } } } } I use Ralph type glass knives for plastic sectioning of biological } } samples. } } } I am experiencing problems in making a proper knife. I have a TAAB } } } histoknifemaker (also says Reichart-Jung). Is there anyone willing to } } offer } } } expertise in this area? } } } } } } -Patrick Brownell } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I don't know the machine, but is TAAB no longer in business in the UK? For the longest time TAAB supplies (altho not generally easily available in the US) was an excellent source of resins and small items. Regardless, I would think that the worn rubber pad is your problem. Again, thinking of experience with other knifemakers, the pads are an essential part of relieving some of the stress in the glass as it breaks. If this pad is uneven, it may not be absorbing the pressure/stress as it should. Perhaps (if TAAB is not around) some other users of the knifemaker might have accessories (like a spare pad) to help you out. Also check the following: depth of the score--if it's too deep or too light, that will affect the break and edge quality; how fast are you breaking after scoring? - a slow break has always been essential to consistently good knife edges for me; check the setting of the clamp(s) and pressure points to ensure that there are no uneven points and that the clamp(s) are providing equal pressure. I can't find the reference right now, but the original publications on making the Ralph knives by hand (it was in Stain Technology, I think) might get you by, and it also might allow you to check the quality of the glass. The technique wasn't too difficult or tedious, and, as I remember (from the dim recesses of ancient history) the percentage of good knives was acceptable. Hope this helps.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Fri, 5 Jan 2001 09:21:24 +0000 (GMT Standard Time), Patton, David wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Perhaps you could get a lab with no problems to make some } knives with your glass and then test them in their lab and } yours (get them to send some of their glass to you as } well to test your knifemaker). That should show if it is } the glass, the knifemaker or the ultramicrotome that is at } fault. } } Dave } } } On Wed, 3 Jan 2001 08:04:10 -0800 "Brownell, Patrick" } {patrick.brownell-at-weyerhaeuser.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } My problem is a poor knife edge. The knife looks fine under casual } } observation, or with small magnification, but when it comes time to actually } } do some sectioning, I get a lot of chattering, or streaking, or both. I } } know make as many test blocks as I do sample blocks, so I don't waste } } samples with all the bad glass knifes. I've gone through several boxes of } } glass over the past two months, and only get a useable knife every 15 to 20 } } breaks. The same knife breaker has worked fine in the past, and I have } } changed the scoring wheel, and checked all that I can. } } } } This knife breaker is also very hard to get parts for. The only obvious } } problem is that one of the pressure points has a rubber pad that is unevenly } } worn. I don't see how that would cause my problem. Again, the knifes look } } good to the naked eye. } } } } The glass I use is from Electron Microscopy Sciences. They are called ultra } } glass knife strips 6.5mm x 25mm x 400 mm. } } } } I guess I'm just looking for someone who has had a similar problem, and } } fixed it in some manner. Could I have gotten a bad batch of glass? I don't } } think it is very likely, I've used several boxes, but they may all be from } } the same batch, for all I know. Is there another brand of glass, or breaker } } that others have found produce reliable knives? } } } } Thanks for any and all help } } } } -Patrick Brownell } } } ---------- } } } From: Katja M. Wolski[SMTP:kwolski-at-hsc.usf.edu] } } } Reply To: kwolski-at-hsc.usf.edu } } } Sent: Wednesday, January 03, 2001 4:26 AM } } } To: Brownell, Patrick } } } Subject: Re: Ralph Glass Knife problems } } } } } } I use a different knife machine, but what's the problem you're having? } } } Are you } } } not getting a good knife edge or what? } } } } } } Katja } } } } } } } } } } } } "Brownell, Patrick" wrote: } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } Hi all! } } } } } } } } I use Ralph type glass knives for plastic sectioning of biological } } } samples. } } } } I am experiencing problems in making a proper knife. I have a TAAB } } } } histoknifemaker (also says Reichart-Jung). Is there anyone willing to } } } offer } } } } expertise in this area? } } } } } } } } -Patrick Brownell } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
On Fri, 5 Jan 2001 05:28:34 -0800 (PST) Roger Moretz {rcmoretz-at-excite.com} wrote:
} Patrick: } } I don't know the machine, but is TAAB no longer in business in the UK? For } the longest time TAAB supplies (altho not generally easily available in the } US) was an excellent source of resins and small items. Regardless, I would } think that the worn rubber pad is your problem. Again, thinking of } experience with other knifemakers, the pads are an essential part of } relieving some of the stress in the glass as it breaks. If this pad is } uneven, it may not be absorbing the pressure/stress as it should. Perhaps } (if TAAB is not around) some other users of the knifemaker might have } accessories (like a spare pad) to help you out. Also check the following: } depth of the score--if it's too deep or too light, that will affect the } break and edge quality; how fast are you breaking after scoring? - a slow } break has always been essential to consistently good knife edges for me; } check the setting of the clamp(s) and pressure points to ensure that there } are no uneven points and that the clamp(s) are providing equal pressure. I } can't find the reference right now, but the original publications on making } the Ralph knives by hand (it was in Stain Technology, I think) might get you } by, and it also might allow you to check the quality of the glass. The } technique wasn't too difficult or tedious, and, as I remember (from the dim } recesses of ancient history) the percentage of good knives was acceptable. } Hope this helps. } } Roger Moretz, Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals, Inc. } } On Fri, 5 Jan 2001 09:21:24 +0000 (GMT Standard Time), Patton, David wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Perhaps you could get a lab with no problems to make some } } knives with your glass and then test them in their lab and } } yours (get them to send some of their glass to you as } } well to test your knifemaker). That should show if it is } } the glass, the knifemaker or the ultramicrotome that is at } } fault. } } } } Dave } } } } } } On Wed, 3 Jan 2001 08:04:10 -0800 "Brownell, Patrick" } } {patrick.brownell-at-weyerhaeuser.com} wrote: } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } My problem is a poor knife edge. The knife looks fine under casual } } } observation, or with small magnification, but when it comes time to } actually } } } do some sectioning, I get a lot of chattering, or streaking, or both. } I } } } know make as many test blocks as I do sample blocks, so I don't waste } } } samples with all the bad glass knifes. I've gone through several boxes } of } } } glass over the past two months, and only get a useable knife every 15 } to 20 } } } breaks. The same knife breaker has worked fine in the past, and I have } } } changed the scoring wheel, and checked all that I can. } } } } } } This knife breaker is also very hard to get parts for. The only } obvious } } } problem is that one of the pressure points has a rubber pad that is } unevenly } } } worn. I don't see how that would cause my problem. Again, the knifes } look } } } good to the naked eye. } } } } } } The glass I use is from Electron Microscopy Sciences. They are called } ultra } } } glass knife strips 6.5mm x 25mm x 400 mm. } } } } } } I guess I'm just looking for someone who has had a similar problem, and } } } fixed it in some manner. Could I have gotten a bad batch of glass? I } don't } } } think it is very likely, I've used several boxes, but they may all be } from } } } the same batch, for all I know. Is there another brand of glass, or } breaker } } } that others have found produce reliable knives? } } } } } } Thanks for any and all help } } } } } } -Patrick Brownell } } } } ---------- } } } } From: Katja M. Wolski[SMTP:kwolski-at-hsc.usf.edu] } } } } Reply To: kwolski-at-hsc.usf.edu } } } } Sent: Wednesday, January 03, 2001 4:26 AM } } } } To: Brownell, Patrick } } } } Subject: Re: Ralph Glass Knife problems } } } } } } } } I use a different knife machine, but what's the problem you're } having? } } } } Are you } } } } not getting a good knife edge or what? } } } } } } } } Katja } } } } } } } } } } } } } } } } "Brownell, Patrick" wrote: } } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } Hi all! } } } } } } } } } } I use Ralph type glass knives for plastic sectioning of biological } } } } samples. } } } } } I am experiencing problems in making a proper knife. I have a TAAB } } } } } histoknifemaker (also says Reichart-Jung). Is there anyone willing } to } } } } offer } } } } } expertise in this area? } } } } } } } } } } -Patrick Brownell } } } } } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } } } _______________________________________________________ } Send a cool gift with your E-Card } http://www.bluemountain.com/giftcenter/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Are you vacuum embedding the glass and Spurr's resin? That should work to get a lot of the void space out.
Just pour the resin into the embedding capsule, put the capsule into a vacuum oven, let pump down for a few minutes (~20 min), bring the sample back to room pressure to let the air pressure force the resin into the pores, then pump it down again and bring back to room pressure and let the sample cure.
dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
} As part of the planned makeover of our building, it has been } proposed ... } Part of that proposal was to set up a computing lab with } fileserver and work-stations networked ... } However, there is great pressure on space in this building, and it } has been suggested to me that computing for imaging and } spectroscopy does not need to be separated from general purpose } computing these days. ...
Somewhat true ... for our EPMA and SEM images, we simply archive to a "files available" location, and every researcher has their own computer and software. However, you would need a workstation for the OM, would you not?
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Email: us004118-at-mindspring.com Name: Leonard Lessin, FBPA
School: (Retired Science & Medical Photographer)
State: NY
Zip: 10012
Question: I am enjoying doing photomicrographs of crystal preperations in polarized light.However I have insuffient knowledge of chemistry to choose solvents without a long series of trial and error efforts.Can you give me a rationale and/or a reference to go about this in a more productive manner?
High quality professional film scanners (LeafScan 45, LS-4500 etc.) are very expansive - over $10k.
I use MINOLTA Dimage Scan Multi (Minolta USA -- how2scan.com ) for digitizing of TEM negatives.
Its features: - Optical resolution: 1128dpi (TEM) 2820dpi (35mm, APS) - Dynamic range: 3.6 - Price: around $2,000
It has one drawback only: TEM film adapter aperture size is of 2.2x3.15. However only central part of a negative is used for image processing.
There is a choice among the price and convenience.
Best regards,
Alexander
Dr. A.S. Solodukhin Department of Anesthesiology University of Virginia Health System P.O. Box 800710 Charlottesville, VA 22906-0710 FAX : (804) 982-0019 Phone: (804) 924-2494
"Dr. Catherine Powell" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are a clinical diagnostic TEM laboratory serving a provincial network of hospitals. We would like to move away from conventional darkroom printing of images and wish to convert negatives to digital files. We are looking for a negative scanner that will: } } 1. Accept 3 X 4 1/4 inch sheet film negatives (Kodak 4489 film). } } 2. Comes with software support that is adaptable to a Windows format. } } 3. Must be priced under $3000. } } 4. Must provide the best quality resolution for diagnostic results (i.e. 3200 X 3600 pixel; wide linear dynamic range). } } I look forward to any information and advice you can provide. } } Catherine Powell } }
Since there has been only one other response, I will toss in my 2 cents.
Some time back I would have said that the load of microscopic images would be more than what the general purpose computing lab would want to take responsibility for. It might prove a significant network traffic load too. However, I think any decent network should be able to handle the load from the microscope labs. On the side, we have a good campus network here at Iowa State. However an analysis of network traffic shows a disproportionate amount of traffic coming out of a few dorm rooms (the normal traffic pattern would be incoming). The university is going to start limiting traffic to so many gigabytes per month or start charging. So I think the microscope load is no longer that significant.
The other issue that might work against it is software licensing. I don't know if you use special programs to work with your images. It might be difficult to work out a suitable licensing arrangement if that software needs to be installed on a lab full of computers, but maybe you can.
As far as regular maintenance of the data, I would be all in favor of letting someone else do the backups providing they have the space (and they should).
Warren
At 06:25 PM 1/2/2001 +0000, you wrote:
} As part of the planned makeover of our building, it has been } proposed that the various and somewhat distributed microscopy } facilities in our building - SEM, TEM, Confocal, fluorescence, } luminescence imaging, darkroom etc. etc. might be brought together } to form a biological imaging facility, thereby benefitting from some } improvements in supervision, security and user support. Part of that } proposal was to set up a computing lab with fileserver and work- } stations networked to the instruments and dedicated to image } storage and off-line image and spectral processing and analysis. } However, there is great pressure on space in this building, and it } has been suggested to me that computing for imaging and } spectroscopy does not need to be separated from general purpose } computing these days. What are your opinions about this? Pro or } con? } } Happy New Millennium } Chris } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr Chris Jeffree } University of Edinburgh
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
we're looking to buy a used carbon evaporator and before I speed through the used equipment websites I thought I'd just check to see if anyone here is thinking of selling their one.
cheers Liz McKenzie
******************************************************* Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
Applications of Scanning Microscopy in Forensic Science
Dear Fellow Microscopist / Forensic Scientist,
SCANNING 2001, the Thirteenth Annual International Scientific Meeting on Scanning Microscopies will be held May 5-7, 2001, in New York City at The Roosevelt Hotel. Please make plans to attend three full days of scientific papers and six short courses all devoted to the use of scanning microscopy. Numerous specific application topics including forensics, electron backscattered diffraction, scanning probe microscopy, nanotechnology, anthropology, modern optical microscopy, museum applications of SEM, food microstructure, material science, microwave techniques and pharmaceuticals. For a completely listing of session topics, short course titles and registration information, please visit the SCANNING web site at www.scanning-fams.org.
CALL FOR PAPERS:
As a co-chair of the SCANNING 2001's "Applications of Scanning Microscopy in Forensic Science" session, I am most pleased with the participation and interest in the forensic session over the last eight years. This year, Mr. Dennis Ward of the Federal Bureau of Investigation, has graciously agreed to co-chair the forensic sessions. Together, we are planning to provide an exciting and informative forensic program. The continued growth of the forensic session over the last eight years will once again permit two full days of forensic papers. The "Applications of Scanning Microscopy in Forensic Science" sessions will be held on Sunday and Monday, May 6-7, 2001. Combined with the popular one day "Scanning Microscopy in Forensic Science" short course (Saturday, May 5, 2001), the forensic scientist/student will be able to attend three consecutive (and full) days of instruction as well as scientific papers on current forensic science research and unique cases all devoted to scanning microscopy applications in forensic science. A large number of microscopy vendors and a full schedule of social activities and tours in New York are also available for a most enjoyable meeting experience.
I encourage you to submit an abstract for platform or poster consideration and be a part of the SCANNING 2001 Forensics Session. Additionally, if you are involved with or know of forensic science students actively engaged in forensic research using any type of scanning microscopy, I strongly encourage you to have your student(s) submit an abstract for consideration as a student paper or poster. Again, please visit the SCANNING web site at www.scanning-fams.org.
Should you have any questions about the forensic symposium, short course or student papers, please feel free to contact me.
See you in New York!
S. Frank Platek, MS Co-Chair, Forensic Symposium and Short Course SCANNING 2001 (513) 679-2700 (513) 679-2761 FAX fplatek-at-ora.fda.gov
Looking for a few good Account Representatives for an electron and optical microscopy service. Territory is open. Excellent earning potential. Call Dick O'Connell at 734-668-3309 or e-mail at oconnell-at-ltu.edu for more information.
I believe TAAB is still in business TAAB Laboratories Equipment Ltd. 3 Minerva House Calleva Park Aldermaston , Berkshire RG7 8NA UK Phone: 44-118-9817775 Fax: 44-118-9817881 E-mail: sales-at-taab.co.uk
Chris
----- Original Message ----- } From: "Roger Moretz" {rcmoretz-at-excite.com} To: "Patton, David" {David.Patton-at-uwe.ac.uk} ; "Brownell, Patrick" {patrick.brownell-at-weyerhaeuser.com} Cc: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, January 05, 2001 1:28 PM
The earliest mention of EM in TV or movies goes back in time much further than "Quincy" or the "X-Files".
I thought I would mention my favorite EM TV/Movie memory. Anyone remember the movie called "The Andromeda Strain" ?? The movie from the 60's was based on a Michael Crighton book by the same name . A Transmission Electron Microscope (RCA EMU ???), as well as "live" images of the ultramicrotomy sectioning process were featured.
Remember?
I would be interested in knowing if there are earlier references than this one.
Best Wishes,
Bill Monroe -- Bill Monroe EM Center Mississippi State University (601)-325-3019 Lab Fax 325-0246
You're correct about the RCA microscope, except that the one in the movie was right after RCA discontinued the business, and it was picked up by "Forgflo", which was the name seen clearly in the movie.
Check it out...
Larry (Master of all trivia...why don't I get on Who Wants to be a Millionaire?) ;-)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The earliest mention of EM in TV or movies goes back in time much } further than "Quincy" or the "X-Files". } } I thought I would mention my favorite EM TV/Movie memory. Anyone } remember the movie called "The Andromeda Strain" ?? The movie from } the 60's was based on a Michael Crighton book by the same name . A } Transmission Electron Microscope (RCA EMU ???), as well as "live" } images of the ultramicrotomy sectioning process were featured. } } Remember? } } I would be interested in knowing if there are earlier references } than this one. } } Best Wishes, } } Bill Monroe } -- } Bill Monroe } EM Center } Mississippi State University } (601)-325-3019 Lab } Fax 325-0246
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
You might also recall that in the Andromeda Strain movie, the specimen was inserted through the viewing port and placed on the fluorescent screen. In this way the were actually able to watch the "virus" replicate right before their eyes.
At 02:39 PM 01/05/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
----- Original Message ----- } From: "Warren E Straszheim" {wesaia-at-iastate.edu} {snip}
} } As far as regular maintenance of the data, I would be all in favor of } letting someone else do the backups providing they have the space (and they } should).
Warren,
If the backups are of any value to you the only person you can trust to make them is yourself. The only method I have found that has never failed me it to compress the files in zip format transfer it to the backup media and verify it on the back up media. Then store the back up media off site. The zip format applies to MSDOS computer other archive file types apply to other OSs.
If the data is really important make two copies.
Unless you can hire a lot better folks then I have seen colleges hire they will make mistakes on backups sooner or later probably sooner.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Of course some of us remember this movie but for heavens sake why did they have green "colored" (interference "color") sections floating in the microtome knife boat when gold surely would have photographed as well and fit the dialogue of that scene. Obviously the technical people just said well cut us some sections to phtograph. A sore point to me at the time.
} } } "William A. Monroe" {monroe-at-emcenter.msstate.edu} 01/05 12:39 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The earliest mention of EM in TV or movies goes back in time much further than "Quincy" or the "X-Files".
I thought I would mention my favorite EM TV/Movie memory. Anyone remember the movie called "The Andromeda Strain" ?? The movie from the 60's was based on a Michael Crighton book by the same name . A Transmission Electron Microscope (RCA EMU ???), as well as "live" images of the ultramicrotomy sectioning process were featured.
Remember?
I would be interested in knowing if there are earlier references than this one.
Best Wishes,
Bill Monroe -- Bill Monroe EM Center Mississippi State University (601)-325-3019 Lab Fax 325-0246
The other fun thing about the crystalline image in that TEM from Andromeda Strain was that it was in color!
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: William A. Monroe [mailto:monroe-at-emcenter.msstate.edu] Sent: Friday, January 05, 2001 3:39 PM To: Warren E Straszheim Cc: Microscopy-at-sparc5.microscopy.com Subject: Re: TV Trivia
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html
The earliest mention of EM in TV or movies goes back in time much further than "Quincy" or the "X-Files".
I thought I would mention my favorite EM TV/Movie memory. Anyone remember the movie called "The Andromeda Strain" ?? The movie from the 60's was based on a Michael Crighton book by the same name . A Transmission Electron Microscope (RCA EMU ???), as well as "live" images of the ultramicrotomy sectioning process were featured.
Remember?
I would be interested in knowing if there are earlier references than this one.
Best Wishes,
Bill Monroe -- Bill Monroe EM Center Mississippi State University (601)-325-3019 Lab Fax 325-0246
"Scanned Probe and Electron Microscopy and Microanalysis: Applications & Techniques" _________________________________________________________________________
The University of Sydney
February 14-16, 2001
AMAS and ASPMS invite you to join us for what will probably be the very first electron and scanned probe microscopy meeting of the 21st Century.
The aim of the Symposium is to provide a forum where participants can discuss electron and scanned probe microscopy, with emphasis on practical solutions and applications. A number of overseas invited speakers will be involved with both the Symposium and the Workshops. We also encourage you to present your recent work using SPM, SEM/TEM and microanalysis.
Pre-Symposium Workshop February 12-13, 2001 Practical Digital Imaging Introduction to SPM Spectral Imaging Materials SPM Environmental SEM Biological SPM Low Voltage SEM and microanalysis SPM of Soft Materials Advances in EBSD in SEM SPM Calibration and Maintenance Introduction to XEDS and EELS in the EM Functionalising Tips
Information concerning the symposium, workshops, accommodation, registration, etc. is posted on our website (www.microscopy.org.au then follow the link to "Upcoming ASPMS, AMAS Symposium". This information will be revised and expanded as necessary.
Just a word of caution about Zip disks. They can be very unreliable. When Iomega first came out with them, their media was really good. Now it is not the same. Maxell and Fujifilm also make media. I think that theirs are better.
The other problem with Iomega drives (Zip and Jaz) is the click of death. This is a precursor to a dead drive or media and loss of all that is on the media (if bad media).
To check your drives and media, run tip.exe. To find out more about this, visit http://www.grc.com
gary g.
At 01:53 PM 1/5/01, you wrote:
} ----- Original Message ----- } } From: "Warren E Straszheim" {wesaia-at-iastate.edu} } {snip} } } } } } As far as regular maintenance of the data, I would be all in favor of } } letting someone else do the backups providing they have the space (and } they } } should). } } Warren, } } If the backups are of any value to you the only person you can trust to } make them is yourself. The only method I have found that has never failed } me it to compress the files in zip format transfer it to the backup media } and verify it on the back up media. Then store the back up media off site. } The zip format applies to MSDOS computer other archive file types apply to } other OSs. } } If the data is really important make two copies. } } Unless you can hire a lot better folks then I have seen colleges hire they } will make mistakes on backups sooner or later probably sooner. } } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
Yes....different "zip." Winzip is very good for munching files into a smaller data space. The media problem remains, however. As you point out, other media keeps coming out.
I tried DVD-RAM and really got whacked by bad sectors. I don't use that any longer. DVD-R is quite another story. CD-R is so cheap these days it is a great way to store 600MB of data. The only problem is to ensure that it remains on the media. Most burner programs do not verify after write. Toast on the Mac does, but Adaptec's programs on the PC do not. I have not found any other programs that will verify after write on the PC. Bummer.
I have not done much at all with CD-RW. I wonder what the experiences have been with this option? My new Yamaha drive is 16x/10x/40x and should do nicely for RW. Never tried it for RW. I guess that I should do that some day. CD-Rs at 12X are iffy....even with certified media. That's why the verification is such an important missing feature.
For really good backup and restore, consider the removable IDE drives. Very nice. My one year's work fits on three 45G drives. Hopefully it remains accessible in the future. Data overload is an emerging problem.
gg
At 08:03 PM 1/5/01, you wrote:
} ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "Gordon Couger" {gcouger-at-couger.com} } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, January 05, 2001 7:16 PM } Subject: Re: Computing for microscopy and imaging } } } } Just a word of caution about Zip disks. They can be very } } unreliable. When Iomega first came out with them, their } } media was really good. Now it is not the same. Maxell } } and Fujifilm also make media. I think that theirs are better. } } } } The other problem with Iomega drives (Zip and Jaz) is the } } click of death. This is a precursor to a dead drive or } } media and loss of all that is on the media (if bad media). } } } } To check your drives and media, run tip.exe. To find out } } more about this, visit http://www.grc.com } } } } gary g. } } Gary, } } Zip files not Zip disk. Use Pkzip or some other program to put all the } files into one zip file, usually a complete directory, copy that file to } the back up media and verify the zip file on the back up media. Currently } I use write only CD-ROMs and keep a copy of really important stuff on my } internet server at my ISP as well. Hard disk space is cheap. I have the } last 15 years work at my finger tips. If I can find it:) I started using } this on single sided single density floppies on a Radio Shack Model 1. I } can only think of two times that I was unable to retrieve a file in 20 } years. } } I am a programmer not a photographer so a years work might fit on a floppy } disk. } } I am not satisfied with the life of CD-ROMs but so far something better } has always come along before the old media went bad and I copied it all } over. One thing I don't like about CD-ROMs is they are not protected from } scratching or being broken. } } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Gordon Couger" {gcouger-at-couger.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, January 05, 2001 10:59 PM
I am looking for accessories and parts for binocular microscope Jenna Technoval 2. Anyone have some spare unneeded accessories as camera attachment, oculars, literature, etc?
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
In one of the Batman movies where he fights the arch villain Mr. Freeze. In the part of the move where the police raid Mr. Freezes' lair you will see off to one side a Philips 400 TEM dressed up as a cheap prop. This is an insult to the Scientist and Technicians who have developed finely honed skills to bring to light the world of the very small in a effort to make life a little better for all of us. Electron Microscopy has been an important part of Science for the last half of the 20th century. This finely engineered tool of Science has open up the world of the very small to eyes that would never have imagined that life could exist on such a level. It sickens me to see this fine and noble tool of Science reduced to a carnival side show gimmick. It is my hope that the men and women of this newsgroup will in their own way and in their own time bring electron microscopy back to its proper place in the scientific community, because if we of this newsgroup and others who labor in Science do not, than this important source of knowledge will be lost to history and the politicians. I will now get off my soap box and make no further comment on the subject.
Ronald Austin Research Associate LSU Medical Center Dept of Pathology Shreveport, LA rla-at-mindspring.com
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-pi.csiro.au] Sent: Friday, January 05, 2001 11:52 PM To: Microscopy-at-sparc5.microscopy.com
And there were all those Zeiss microscopes in Jurassic Park. Pity the one they were using in one scene (an Axiophot?) didn't have a condenser....
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Ah, come on. There were many technical errors in that film, but wasn't it fun? One of my favorite faux pas was the rapid growth? multiplication? of the strain. Not to mention it's apparent ability to degrade a variety of natural and synthetic rubbers while also attacking human blood plasma.
Michael Crichton did a good job of describing a possible protagonist, and Hollyweird did its usual job of visualizing it. To expect anything different would be folly.
On Friday, January 05, 2001 4:59 PM, Walck, Scott D. [SMTP:walck-at-ppg.com] wrote: } } The other fun thing about the crystalline image in that TEM from Andromeda Strain was that it was in color! } } } -Scott } } -----Original Message----- } From: William A. Monroe [mailto:monroe-at-emcenter.msstate.edu]
} } The earliest mention of EM in TV or movies goes back in time much } further than "Quincy" or the "X-Files". } } I thought I would mention my favorite EM TV/Movie memory. Anyone } remember the movie called "The Andromeda Strain" ?? The movie from } the 60's was based on a Michael Crighton book by the same name . A } Transmission Electron Microscope (RCA EMU ???), as well as "live" } images of the ultramicrotomy sectioning process were featured. } } Remember? } } I would be interested in knowing if there are earlier } references than this one. } } Best Wishes, } } Bill Monroe } -- } Bill Monroe } EM Center } Mississippi State University } (601)-325-3019 Lab } Fax 325-0246 } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} } Yes....different "zip." Winzip is very good for munching } } files into a smaller data space. The media problem remains, } } however. As you point out, other media keeps coming out. } ============================ } I really think the best media out there for long term storage would be } plain old 35 mm film. I don't know what kind of data density you could get } on tech pan but I would trust the data to be there when I wanted to get it } back.
Winzip does a good job on data that is compressible. Non-LZW TIFF can be compressed as can ASCII and other byte hungry data. But I don't find much size reduction with compressed TIFF or JPEG. These are mostly what I have. Even then, my native images are uncompressed TIFF. Those are what I need to save as backups plus backups of the backups.
} } } } I have not done much at all with CD-RW. I wonder what } } the experiences have been with this option? My new Yamaha } } drive is 16x/10x/40x and should do nicely for RW. Never tried } } it for RW. I guess that I should do that some day. CD-Rs at } } 12X are iffy....even with certified media. That's why the } } verification is such an important missing feature. } =============== } } From the reports I see on RW CDROMs the storage life is a lot shorter than } a R CDROM. Both of them use some kind of dye process and the RW is } reversable.
I too have not heard good things about RW. Unless and until the viability is assured, my precious data isn't going on an RW media. When its gone, its gone. Not a good situation.
} It takes time and markting didn't like it is my guess. But that is why you } have to verify each file you copy with verify option of Winzip. It reads } the file and does the CRC and checks it against the one in the file. } } It is slow and cumbersom but it is the only way I have found that works on } MSDOS or Windos. On Linux or Unix I can write a script to do it all. I } have been involved in disaster recovereries and most of the times at least } part of the back ups are bad. Some of the restore system can't recover } from a bad file. I don't know much about backup systems for contemperary } systems I just copy every thing to a CDROM uncompressed and put a } compressed version on the network box and call it good.
Hum. I'm using Winzip 7 and it is very fast on Win98SE. Stuffit on the Mac also does a nice job.
} } } } For really good backup and restore, consider the removable } } IDE drives. Very nice. My one year's work fits on three 45G } } drives. Hopefully it remains accessible in the future. Data } } overload is an emerging problem. } ================================== } That's what I do on my Linux box on the internet. I have a drive for } backups. Drives are cheap. High quality digital imagining is going to } really eat up disk space. We need 10 or 20 gig CDROMs
Yes indeed. Even a 10G CD would be great, if it worked. I've toyed with the idea of DVD-R which will do 4.7G and 9G. But the drives are very pricey as is the media. And I suspect they may have the same reliability issues as CD-RW and even CD-R. Not all CD-R writes work 100%. Only Toast on the Mac will do a verify after write. I've not found any burner program for the PC that will verify after write. Strange.
The IDE drive trays seem like a good approach to backup. The drives too are obsoleted rather quickly. My "new" IBM 45G Deskstar ATA-66 drives are now discontinued. IBM makes ATA-100 45G drives for $239 versus the $129 I paid for the ATA-66 ones.
My only other backup option (a redundant one) is a Sony SDT-9000 4mm DAT. Using native hardware compression, it will store 24G on a DDS-3 120meter tape. But this has some peculiar problems based on lack of backup software for tape that works in a dual host adapter environment. Dantz's Retrospect works great on the Mac but fails on the PC. I have to use old Win95 Adaptec backup. It works but does not offer user interface to the drive's features.
It seems that as we rush from film to digital, we may look back on the thousands of negs or chromes sitting in file cabinets and wonder what happened. We'd now have file cabinets of $200 hard drives which may or may not work after sitting for some amount of time. A rather unsettling feeling.
Holey Microscopy Batman...maybe we should blame that on Alfred. And to think I didn't tune in the following night...same bat time...same bat channel...to see if this oversight was corrected.
-----Original Message----- } From: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Saturday, January 06, 2001 12:18 AM To: Microscopy Society of America
In one of the Batman movies where he fights the arch villain Mr. Freeze. In the part of the move where the police raid Mr. Freezes' lair you will see off to one side a Philips 400 TEM dressed up as a cheap prop. This is an insult to the Scientist and Technicians who have developed finely honed skills to bring to light the world of the very small in a effort to make life a little better for all of us. Electron Microscopy has been an important part of Science for the last half of the 20th century. This finely engineered tool of Science has open up the world of the very small to eyes that would never have imagined that life could exist on such a level. It sickens me to see this fine and noble tool of Science reduced to a carnival side show gimmick. It is my hope that the men and women of this newsgroup will in their own way and in their own time bring electron microscopy back to its proper place in the scientific community, because if we of this newsgroup and others who labor in Science do not, than this important source of knowledge will be lost to history and the politicians. I will now get off my soap box and make no further comment on the subject.
Ronald Austin Research Associate LSU Medical Center Dept of Pathology Shreveport, LA rla-at-mindspring.com
I have enjoyed reading all the TV trivia regarding the Quincy and Andromeda Strain. What many of you may not know is that for the "Strain" movie the makers of the film contacted a now defunct Microprobe Company, Materials Analysis Co. We sent to the studio an Electron Microprobe. The only portion that made the film was some blinking x-ray scalers and a portion of the electronics rack. What the impact of microprobe was in the film I can't guess or remember. In the Quincy episodes, there were more than one, we sent to the studio an ISI SEM along with a service engineer, whose name at the moment escapes me, to install it and our application and Demonstration man, Bill Roth to run the SEM, Bill has been with Hitachi for many years now and could give more incite as to what happened at the studio than I can as he was there for a week doing the one episode. I remember him telling me that Quincy's technician, Sam was explaining to Quincy what was on the CRT with the camera going from the control panel with Bill's hands being filmed but the overall shot of Sam and Quincy looking at the CRT and discussing the forensic material. I still have some 8x10's around here somewhere of the filming. Thought you might be interested.
{snip} } Winzip does a good job on data that is compressible. Non-LZW TIFF } can be compressed as can ASCII and other byte hungry data. But } I don't find much size reduction with compressed TIFF or JPEG. These } are mostly what I have. Even then, my native images are uncompressed } TIFF. Those are what I need to save as backups plus backups of the } backups. ========= These files are already compressed as much as possible. The only advantage of zip programs they get them in one file. {snip} } } } For really good backup and restore, consider the removable } } } IDE drives. Very nice. My one year's work fits on three 45G } } } drives. Hopefully it remains accessible in the future. Data } } } overload is an emerging problem. } } ================================== } } That's what I do on my Linux box on the internet. I have a drive for } } backups. Drives are cheap. High quality digital imagining is going to } } really eat up disk space. We need 10 or 20 gig CD-ROMs } } Yes indeed. Even a 10G CD would be great, if it worked. I've } toyed with the idea of DVD-R which will do 4.7G and 9G. But } the drives are very pricey as is the media. And I suspect they } may have the same reliability issues as CD-RW and even CD-R. } Not all CD-R writes work 100%. Only Toast on the Mac will do } a verify after write. I've not found any burner program for the PC } that will verify after write. Strange. } } The IDE drive trays seem like a good approach to backup. The } drives too are obsolete rather quickly. My "new" IBM 45G Deskstar } ATA-66 drives are now discontinued. IBM makes ATA-100 } 45G drives for $239 versus the $129 I paid for the ATA-66 ones. ============= I just use a regular IDE drive it's about as cheap and not much more trouble to change if you put it in the bottom bay and don't screw it in. That works well for me but it would not work well for large images. They just generate too much data. } } My only other backup option (a redundant one) is a Sony SDT-9000 } 4mm DAT. Using native hardware compression, it will store 24G } on a DDS-3 120meter tape. But this has some peculiar problems } based on lack of backup software for tape that works in a dual } host adapter environment. Dantz's Retrospect works great on } the Mac but fails on the PC. I have to use old Win95 Adaptec } backup. It works but does not offer user interface to the } drive's features. ==================== Be careful reusing tapes. They can only be use a few times before they start getting unreliable. One I was using recommended 5 reuses.
Tape is also slow to get a single file from. The one I want is always on the end. } } It seems that as we rush from film to digital, we may look back } on the thousands of negs or chromes sitting in file cabinets } and wonder what happened. We'd now have file cabinets of } $200 hard drives which may or may not work after sitting for } some amount of time. A rather unsettling feeling.
I agree that it is unsettling. That why I have said in the past photographic film is probably still the best archival media we have from cost, aging and resolution point of view. It's fatal flaw is lack of instant availability and knowing if you got a good shot or not while you are still set up.
An interesting setup would be a beam splitter that would let you take either plain old film or CCD images. Then you could do your long term storage on film that we know won't go obsolete with of the next upgrade and we have no questions about how long it will last. Film is competitive or cheaper with digital storage if you include resolution in the equation.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
What I have been doing, for convenience more than anything, is to copy my SEM & PM prints with a digital camera at 1024 by 768 resolution. I use a copy stand for its light sources but I hand-hold the camera (!). After all, the images are already at their maximum enlargement, so I won't be looking for more detail than can already be seen in the original image. They print at 200 dpi, not as good as what the printer can do (600 dpi) but they look fine in my reports, and then I can use HTML to write the report because the macro images are in a digital format, too. Extra copies are no longer a problem; in the "good old days" we (Amenex's microscopist, that is) had to spend days at a time making contact prints in a rube-goldberg photo lab set up in the metallographic preparation room (only one that's dark enough).
On the other hand, when one starts making digitally recorded images on the SEM, theoretically one is creating a super-wide-angle image. Does the resolution hold up ? If not, we're kidding ourselves like the fly on a raft who wants the drawbridges raised ...
Best regards, George Langford, Sc.D. amenex-at-amenex.com http://www.amenex.com/
I would greatly appreciate some hints concerning the kind of materials (minerals ?) that could be used in order to determine the k(A,B) (Cliff-Lorimer approx.) factors for my instrument (TEM CM-200 Philips microscope with EDAX EDS X-ray spectrometer) concerning Zr(K-lines), Y(K-lines), Hf(L-lines) and La(?-lines). Do they exist Si containing minerals which could be used to determine the k factors relating Si and the above mentioned elements ? Are they suppliers of standard materials that could be used for determining the k factors for the above mentioned elements ?
Thank you in advance !
Corneliu Sarbu, PhD Dept.of Metallurgy and Applied Materials Science Catholic University of Leuven Belgium
Email: jwahom01-at-tufts.edu Name: Jane Wahome School: Tufts University
Question: I need to view calcium carbonate crystals with a transmission electron microscope for a research project. I also need to record the images. All I've managed to do so far is order thin carbon grids. I have been asked to write a plan for my experiment but I have no clue how to prepare crystals in solution for magnification or what tools to use. Please give me some direction. My professors say I should figure everything out myself - "It will be a good learning experience."
I only do digital images on the SEM. There are several providers of this capability for retrofitting. Some if not most all current SEM makers include digital imaging. When running an active scan system, the digital image is just as on the TV screen and Polaroid output film. But it can be made different. Since the voltage points for limits of the scanned frame are user programmable, a small region can be scanned at very high resolution or a larger region at a lower resolution. The limiting factor is the number of bits in the D/A converter which drives the scan coils. Most of these are 12-bits, so that makes 4096 discrete positions across the X range specified and 4096 discrete positions for the Y range.
My highest recording setting is 4032x2688 pixels which is 10.8M pixels at 8-bits per pixel or twice that at 16-bits per pixel. The 4032x2688 values are chosen to make the aspect ratio 1.5:1, so it fully fits a 35mm frame. The normal 1.33:1 does not. Given that the scan limits are set as for a Polaroid shot, the equivalent pixel density is between 700 and 800 pixels per inch for the digital capture image. Not bad at all. And since the pixel dwell time is programmable, one can optimize it for scan time and minimum noise.
gary g.
At 06:12 AM 1/7/01, you wrote: } Hello Gary, Gordon & Microscopists ! } } [snip] } On the other hand, when one starts making digitally } recorded images on the SEM, theoretically one is creating } a super-wide-angle image. Does the resolution hold up ? } If not, we're kidding ourselves like the fly on a raft } who wants the drawbridges raised ... } } Best regards, } George Langford, Sc.D. } amenex-at-amenex.com } http://www.amenex.com/
----- Original Message ----- } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: "Gordon Couger" {gcouger-at-couger.com} Cc: {microscopy-at-sparc5.microsocpy.com} Sent: Sunday, January 07, 2001 8:34 PM
George, What do you mean by a super-wide-angle picture? The digital scan output goes through the mag control, so your angle is the same as for Polaroid at a given mag. The resolution is determined by the pixel density and won't exceed a Type 55 negative on your ETEC until you capture about 4k by 4k. 2k by 2k isn't quite as good as you can do with film, the limiting factor being your record CRT and camera. As Gordon said, film is still the measure. It lasts, the resolution is great and software "upgrades" won't make your entire file obsolete.
Besides, don't you have some problems using pure digital imaging for legal cases? My understanding is that if you use digital, the original file must be on a WORM drive (now known as CD-R) to give the equivalent of an original negative on file. This info came from the Virginia Crime Labs quite a few years ago.
Ken Converse, owner Quality Images Delat, PA
George Langford, Sc.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Gary, Gordon & Microscopists ! } } What I have been doing, for convenience more than anything, } is to copy my SEM & PM prints with a digital camera at 1024 } by 768 resolution. I use a copy stand for its light sources } but I hand-hold the camera (!). After all, the images are } already at their maximum enlargement, so I won't be looking } for more detail than can already be seen in the original } image. They print at 200 dpi, not as good as what the printer } can do (600 dpi) but they look fine in my reports, and then } I can use HTML to write the report because the macro images } are in a digital format, too. Extra copies are no longer a } problem; in the "good old days" we (Amenex's microscopist, } that is) had to spend days at a time making contact prints } in a rube-goldberg photo lab set up in the metallographic } preparation room (only one that's dark enough).
} } On the other hand, when one starts making digitally } recorded images on the SEM, theoretically one is creating } a super-wide-angle image. Does the resolution hold up ? } If not, we're kidding ourselves like the fly on a raft } who wants the drawbridges raised ... } } Best regards, } George Langford, Sc.D. } amenex-at-amenex.com } http://www.amenex.com/ } } }
} What do you mean by a super-wide-angle picture? The digital scan } output goes through the mag control, so your angle is the same as } for Polaroid at a given mag. The resolution is determined by the } pixel density and won't exceed a Type 55 negative on your ETEC } until you capture about 4k by 4k. 2k by 2k isn't quite as good as } you can do with film, the limiting factor being your record CRT and } camera.
The image I see on the Polaroid film comes nowhere near taxing the resolution of the film. My 1024 by 768 pixel digicam snapshots of the original Polaroid prints look pretty close to the originals. Why enlarge them any more ? Empty magnification and all that. Yes, I know that the twelve- or sixteen-bit ADC's can capture a much wider range of exposure than can film, but I don't see where all those pixels get us any more spatial resolution. A 35 mm camera can cram a lot of resolution onto a small area of film, but its lens is reducing the original scene, not enlarging it. All the fancy lenses on our metallographs can't do any better than covering the 4X5 inch format of the film at maximum resolution. If we try to use a smaller eyepiece to get a wide-angle effect, then the edges of the image show terrible distortion. So there's no point in using an excessive number of pixels to describe the output of a microscope's imaging system.
} As Gordon said, film is still the measure. It lasts, the } resolution is great and software "upgrades" won't make your entire } file obsolete.
Isn't it the hardware that goes South ? Where do I read my eight-inch floppies; or the 5-1/4 inch ones, for that matter ... ? Once a bit map, always a bit map, I should think. I do notice that my .JPG image editor can't make heads or tails of some of the .JPG files that look fine on Netscape, so there are some software issues, but it looks as though the old drives are the real problem.
} Besides, don't you have some problems using pure digital imaging } for legal cases? My understanding is that if you use digital, the } original file must be on a WORM drive (now known as CD-R) to give } the equivalent of an original negative on file. This info came from } the Virginia Crime Labs quite a few years ago.
I lock the original floppy disk and leave the original image files untouched there. Then I copy 'em to my hard drive for cropping and adjustment of contrast, etc. Anyone wants to see the report, gets the whole shebang: originals, thumbnails, edited pix, and text (HTML). CDROM's are handy for the monster files that result. One report had almost a thousand individual files in it. Client wasn't too happy with the task of learning what a browser is. Printing it all out is a monumental PIA, but then, so is ruffling through a two-inch-thick pile of prints.
Best regards, George Langford Principal Consultant Amenex Associates, Inc. http://www.amenex.com/
I can assure you all that TAAB is well and truly still in business as we communicate on a regular basis concerning cryostats and microtomes. Please contact TAAB on: sales-at-taab.co.uk
Best. Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England
Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: AlanBright-at-brightinstruments.com Web Site: www.brightinstruments.com
-----Original Message----- } From: Roger Moretz [mailto:rcmoretz-at-excite.com] Sent: 05 January 2001 13:29 To: Patton, David; Brownell, Patrick Cc: 'Microscopy-at-MSA.Microscopy.Com'
Patrick:
I don't know the machine, but is TAAB no longer in business in the UK? For the longest time TAAB supplies (altho not generally easily available in the US) was an excellent source of resins and small items. Regardless, I would think that the worn rubber pad is your problem. Again, thinking of experience with other knifemakers, the pads are an essential part of relieving some of the stress in the glass as it breaks. If this pad is uneven, it may not be absorbing the pressure/stress as it should. Perhaps (if TAAB is not around) some other users of the knifemaker might have accessories (like a spare pad) to help you out. Also check the following: depth of the score--if it's too deep or too light, that will affect the break and edge quality; how fast are you breaking after scoring? - a slow break has always been essential to consistently good knife edges for me; check the setting of the clamp(s) and pressure points to ensure that there are no uneven points and that the clamp(s) are providing equal pressure. I can't find the reference right now, but the original publications on making the Ralph knives by hand (it was in Stain Technology, I think) might get you by, and it also might allow you to check the quality of the glass. The technique wasn't too difficult or tedious, and, as I remember (from the dim recesses of ancient history) the percentage of good knives was acceptable. Hope this helps.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Fri, 5 Jan 2001 09:21:24 +0000 (GMT Standard Time), Patton, David wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Perhaps you could get a lab with no problems to make some } knives with your glass and then test them in their lab and } yours (get them to send some of their glass to you as } well to test your knifemaker). That should show if it is } the glass, the knifemaker or the ultramicrotome that is at } fault. } } Dave } } } On Wed, 3 Jan 2001 08:04:10 -0800 "Brownell, Patrick" } {patrick.brownell-at-weyerhaeuser.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } My problem is a poor knife edge. The knife looks fine under casual } } observation, or with small magnification, but when it comes time to actually } } do some sectioning, I get a lot of chattering, or streaking, or both. I } } know make as many test blocks as I do sample blocks, so I don't waste } } samples with all the bad glass knifes. I've gone through several boxes of } } glass over the past two months, and only get a useable knife every 15 to 20 } } breaks. The same knife breaker has worked fine in the past, and I have } } changed the scoring wheel, and checked all that I can. } } } } This knife breaker is also very hard to get parts for. The only obvious } } problem is that one of the pressure points has a rubber pad that is unevenly } } worn. I don't see how that would cause my problem. Again, the knifes look } } good to the naked eye. } } } } The glass I use is from Electron Microscopy Sciences. They are called ultra } } glass knife strips 6.5mm x 25mm x 400 mm. } } } } I guess I'm just looking for someone who has had a similar problem, and } } fixed it in some manner. Could I have gotten a bad batch of glass? I don't } } think it is very likely, I've used several boxes, but they may all be from } } the same batch, for all I know. Is there another brand of glass, or breaker } } that others have found produce reliable knives? } } } } Thanks for any and all help } } } } -Patrick Brownell } } } ---------- } } } From: Katja M. Wolski[SMTP:kwolski-at-hsc.usf.edu] } } } Reply To: kwolski-at-hsc.usf.edu } } } Sent: Wednesday, January 03, 2001 4:26 AM } } } To: Brownell, Patrick } } } Subject: Re: Ralph Glass Knife problems } } } } } } I use a different knife machine, but what's the problem you're having? } } } Are you } } } not getting a good knife edge or what? } } } } } } Katja } } } } } } } } } } } } "Brownell, Patrick" wrote: } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } Hi all! } } } } } } } } I use Ralph type glass knives for plastic sectioning of biological } } } samples. } } } } I am experiencing problems in making a proper knife. I have a TAAB } } } } histoknifemaker (also says Reichart-Jung). Is there anyone willing to } } } offer } } } } expertise in this area? } } } } } } } } -Patrick Brownell } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
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We are using Magneto-Optical Disks, 2.6 or 5.2 Gb to store the data. They are very reliable (manufacturer claims, that data may be stored up to 50 years) it's not sensitive for magnetic fields and heat (in reasonable temperature diapason, actually until melted). You may rewrite data many times (a million, I believe). Because of reliability, US government uses this media to store all digital data. MO disk needs special drive, which suppose to be connected to the computer (to the PC in our case, SCSII). It's connected to the network, so it is accessible from other computers. We have to insert/remove MO disk by hands of coarse. 2.6 Gb MO is about $40-80 each. MO drive - about $1000. I am not so happy with this instrumentation (relatively slow, but faster than CD or Zip-drive, you have to have special MO-drive connected to the particular computer etc), but it's only known to me a media, which is reliable: CDR/CDRW - absolutely not reliable, light-sensitive, etc.; Zip-drive/diskette is not enough for me, HD- not bad idea, but technically not that easy (you have to have removable HD, I did have a problem with that setup when WinNT changed the letters to the logical drives and therefore all my programs suddenly stopped when I removed the HD, and you have to restart the computer if you want to remove HD). Currently, I do have approximately 10 Gb data stored on MO disks. In our Department there are 200 or so disks are in use. To my knowledge we did have one case when MO disk virtually lost the data but all 100% data has been recovered later. Something like that may happens if you will try to remove the disk during the writhing (what, probably was happens). If I am going to work with some block of data, I copied that to the server and work on it from any computer even from home. The same happens with fresh portion of data: I temporary store the data on the server and then (after frequent reminding from SysAdmin), transfer it to the MO disk. It takes approximately 20 min to transfer 1 Gb of data. Detailed information about MO disks you may find on the Internet.
To help improve the reliability of ZIP disk there is a freeware program available from Gibson Research, TIP.exe . see http://www.grc.com
At 08:16 PM 1/5/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } And there were all those Zeiss microscopes in Jurassic Park. Pity the one } they were using in one scene (an Axiophot?) didn't have a condenser.... } And let's not forget the Cambodian lady in "Blade Runner" (1982) who operates an SEM in the street (in the rain!!?). She helps Harrison Ford identify a scale in evidence as having come from a bioengineered snake, not a fish (bioengineered animals apparently have serial numbers on even their smallest parts). There was another, more recent movie called "Mimic" in which there is a scene where an insect specialist has a pile of SEM micrographs on her desk, purportedly representing various insect eggs or larvae. As a former micropaleontologist, I could tell they were actually illustrations of planktonic Foraminifera, but I suspect this distinction was lost on much of the viewing public.
Frank Thomas Geological Survey of Canada Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
I have done a lot of EM on ZnO and CaCO3 crystals a while ago. I simply made a suspension of the powder in ethanol or methanol and ultrasonicated them for 3 to 5 min. I then just dropped one or two drops of the suspension on the grid positioned on a filter paper and let it dry. You might want to check with the SEM that you do not change morphologies by the ultrasound treatment. I suggest that you first take some SEM images anyway to get some information on sizes, distributions and morphologies. IF you have rather large crystals. SEM will not work, if the crystals are too small. In that case you will have to go directly to the TEM. If you are taking the crystals directly from a reaction solution, just give a drop or two on the grid and let dry. Of course in that case, you might also have non CaCO3 material on the grid depending on your reaction. You will have to make sure that you are looking at the CaCO3 and not some other crap. This can be done by electron diffraction or HRTEM. Image recording possibilities depend on your microscope. What microscope are you using ? If you need further information or want to discuss some things feel free to send an email.
Andreas
************************************************* Dr. Andreas Taubert Materials Science and Engineering Dept. 3231 Walnut Street The University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
----- Original Message ----- } From: {jwahom01-at-tufts.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, January 07, 2001 11:23 AM
Or not. I guess I just can't ignore this thread. I have always been vocal about inconsistencies/stupidities/errors in TV/movies/etc. So, as to Andromeda Strain: first of all, there was the insertion of the specimen without use of an airlock (and those of us who suffered with the Forgflo/nee RCA EMU-4 know that the beast had this horrid airlock that used the external bellows minipump!!!); then there was the immediate location of the desired area under the beam....; plus all those already mentioned. On Quincy: the first episode (or maybe 2) the lab was fully outfitted with Zeiss equipment; the rest of the series the lab was outfitted with AO (must be nice to have that kind of equipment budget--but why go that direction??? could it be politics???); then the SEM/microprobe episode, where, once again, the area of interest with the decisive inclusion was magically right in the area being examined immediately; then there was the TEM episode where Sam received the biopsy about 4am and had a block, stained sections and a confirmatory diagnosis by 8am (now there's a reality check for you--did your boss pick up on that and demand that turn-around for you????); I'm sure there were others but those stuck (mostly in my craw). And a final overall plaint about the fact that everybody was working (at high mag, no less) in brightly lit rooms, when we toiled away in near pitch dark!! Even that Forgflo/RCA with its ma beam current couldn't do that--I know from many hours in the dark, dark adjusting my eyes.
O well, as someone put it--that's Hollyweird.
Roger Moretz Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Sat, 06 Jan 2001 11:19:43 -0800, Robert Ruscica wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have enjoyed reading all the TV trivia regarding the Quincy and Andromeda } Strain. What many of you may not know is that for the "Strain" movie the } makers of the film contacted a now defunct Microprobe Company, Materials } Analysis Co. We sent to the studio an Electron Microprobe. The only portion } that made the film was some blinking x-ray scalers and a portion of the } electronics rack. What the impact of microprobe was in the film I can't } guess or remember. } In the Quincy episodes, there were more than one, we sent to the studio an } ISI SEM along with a service engineer, whose name at the moment escapes } me, to install it and our application and Demonstration man, Bill Roth to } run the SEM, Bill has been with Hitachi for many years now and could give
} more incite as to what happened at the studio than I can as he was there } for a week doing the one episode. I remember him telling me that Quincy's
} technician, Sam was explaining to Quincy what was on the CRT with the } camera going from the control panel with Bill's hands being filmed but the } overall shot of Sam and Quincy looking at the CRT and discussing the } forensic material. I still have some 8x10's around here somewhere of the } filming. Thought you might be interested. } } Regards, } Bob Ruscica } }
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I need some assistance in making up a uranyl acetate stain. I have powdered uranyl acetate and would like to know what concentration and solvent to use to make a good stain for looking at meat samples in the TEM. If there are any procedures (time and temp) that I should adhere too that would also be much appreciated.
Ps. I'm microtoming the TEM samples first then applying the uranyl acetate stain with a follow-up stain of RuO4 vapor.
thanks all dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
Gatan, Inc the technology leader in digital imaging, analytical spectroscopy and ion beam milling applications for electron microscopy is seeking candidates for an Analytical Product Manager. This person will manage the complete product life cycle for this company's product line. Responsibilities include coordinating all activities necessary to achieve the strategic revenue and profit objectives for the product. For the Product manager there are three key product cycle phases: 1) Planning: includes market research, Marketing plan production and results in a vision statement to pass to development. This the key stage in the development of any new product. The vision statement is the marketing vision for the product and it may include an analysis of competitor's products and a projection of opportunities in the future. 2) Development: Pricing, product positioning, product packaging and product promotion.3) Stabilization: Beta testing, code testing and feedback to developers and product launch. The applicant must have significant product management experience, preferably in a product area that is applicable to market: TEM applications, electron microscopy, biological, materials or semiconductor experience would be useful. Familiarity with existing vendors, consultants, competitors, etc in the industry is a plus. Technical knowledge of Gatan software and hardware applications. A proven track record in both planning and executing successful product management programs. Must be comfortable with current development environments and development tools to work intelligently with engineering. Must have excellent overall PC computing skills, including a thorough familiarity with market tools in document composition, database design and presentation management. MBA preferred; PHD desired with a background in TEM and experience with GIF and PEELS systems. Salary: Base salary plus bonus commensurate with experience. Interested candidates should send email or fax their resume to:
GATAN, INC Attn: HR Department 5933 Coronado Lane Pleasanton, CA. 94588 Fax: (925) 463-0204 Email: hr-at-gatan.com www.gatan.com
Carlotta Daniels GATAN, Inc. Human Resources Pleasanton, CA. 94583
there are also a number of follow-up links, which I have not explored.
The latter link has the following section:
So, How Long Can CDs Last? Leaving aside scratches, fires, floods, and peanut butter sandwiches and concentrating on the slow chemical changes that determine the inherent life expectancy of a CD, extensive accelerated-aging tests suggest that Kodak writable CD products, including Photo CD discs, will not reach a BLERmax of 50 for a period of around 200 years when kept in the dark at moderate storage conditions. This long potential life expectancy is mainly a function of the greater dark stability of the dye used in Kodak writable CD products. Considering that BLERmax 50 is still not an unreadable level of error, Kodak writable CDs have a very long life expectancy indeed. Similar research by the 3M Company shows that CD-ROM products made by them will not attain a block error rate of 50 per second for more than 100 years in moderate storage conditions.
Accelerated aging is subject to uncertainties, but it does rest on firm scientific footing. Behind the data is the simple assumption that raising the temperature causes the reactions of decay to happen faster--so fast, in fact, that they occur within a few months, rather than decades. The science of reaction rates is called kinetics, and the lifetime predictions are based on well-established principles of that branch of chemical science. These same principles are used every day to design the chemical plants and processes of the modern world. Because there is so much practical experience with the laws of kinetics, lifetime predictions based on them are approximately correct. Such test methods soon will be part of a forthcoming ANSI (American National Standards Institute) standard dealing with tests for CD permanence.
End of citation.
So, from a scientific standpoint (accelerated aging), a good writeable CD should have a lifespan of 100-200 years, which is more than tape (several decades) and on par with film, at least as far as I can tell. Film suffers from the same problems the CD suffer from (sensitivity to light, scratches, fire, alien attacks, etc.).
Finally a few words regarding M/O and CD:
We supplied with our systems until a few years ago an M/O drive. At that time, there were only CD-ROMs. The drives were (are?) fairly expensive, about $1000 for the drive, and with the advent of cheap CD-Rs it became harder and harder to find drives and media. The drives are much more common in Europe. In addition, the drives are pretty slow, and it was always a hassle to work with the drives under Windows NT. The drives use a laser to heat the storage medium, then a magnetic head to write the info to the heated patch. This might result in a more stable storage than CD or magnetic. The availability, however, seems to be a big problem.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Monday, January 08, 2001 4:08 AM To: microscopy-at-sparc5.microscopy.com
Hello,
We are using Magneto-Optical Disks, 2.6 or 5.2 Gb to store the data. They are very reliable (manufacturer claims, that data may be stored up to 50 years) it's not sensitive for magnetic fields and heat (in reasonable temperature diapason, actually until melted). You may rewrite data many times (a million, I believe). Because of reliability, US government uses this media to store all digital data. MO disk needs special drive, which suppose to be connected to the computer (to the PC in our case, SCSII). It's connected to the network, so it is accessible from other computers. We have to insert/remove MO disk by hands of coarse. 2.6 Gb MO is about $40-80 each. MO drive - about $1000. I am not so happy with this instrumentation (relatively slow, but faster than CD or Zip-drive, you have to have special MO-drive connected to the particular computer etc), but it's only known to me a media, which is reliable: CDR/CDRW - absolutely not reliable, light-sensitive, etc.; Zip-drive/diskette is not enough for me, HD- not bad idea, but technically not that easy (you have to have removable HD, I did have a problem with that setup when WinNT changed the letters to the logical drives and therefore all my programs suddenly stopped when I removed the HD, and you have to restart the computer if you want to remove HD). Currently, I do have approximately 10 Gb data stored on MO disks. In our Department there are 200 or so disks are in use. To my knowledge we did have one case when MO disk virtually lost the data but all 100% data has been recovered later. Something like that may happens if you will try to remove the disk during the writhing (what, probably was happens). If I am going to work with some block of data, I copied that to the server and work on it from any computer even from home. The same happens with fresh portion of data: I temporary store the data on the server and then (after frequent reminding from SysAdmin), transfer it to the MO disk. It takes approximately 20 min to transfer 1 Gb of data. Detailed information about MO disks you may find on the Internet.
} } Could someone post the reference for the article for making glass knives } by hand?
Here's the original reference, which includes a bit of the history of Ralph knives (in honor of the late Dr. Paul Ralph): Stain Technology 51(2): 71-97. [1976]. Bennet et al. Science and art in preparing tissues embedded in plastic for light microscopy, with special reference to glycol methacrylate, glass knives and simple stains.
Good luck. Mike Nesson _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu
Out here in Sunny Southern California we just purchased a AMT 1K digital camera for use in our EM lab at the UCLA Medical Center. The storage device or choice for archiving we utilize is to burn CD's of the images as long term storage.
=========== } } Just a word of caution about Zip disks. They can be very } } unreliable. When Iomega first came out with them, their } } media was really good. Now it is not the same. Maxell } } and Fujifilm also make media. I think that theirs are better. } } } } The other problem with Iomega drives (Zip and Jaz) is the } } click of death. This is a precursor to a dead drive or } } media and loss of all that is on the media (if bad media). } } } } To check your drives and media, run tip.exe. To find out } } more about this, visit http://www.grc.com } } } } gary g. } } Gary, } } Zip files not Zip disk. Use Pkzip or some other program to put all the } files into one zip file, usually a complete directory, copy that file to } the back up media and verify the zip file on the back up media. Currently } I use write only CD-ROMs and keep a copy of really important stuff on my } internet server at my ISP as well. Hard disk space is cheap. I have the } last 15 years work at my finger tips. If I can find it:) I started using } this on single sided single density floppies on a Radio Shack Model 1. I } can only think of two times that I was unable to retrieve a file in 20 } years. } } I am a programmer not a photographer so a years work might fit on a floppy } disk. } } I am not satisfied with the life of CD-ROMs but so far something better } has always come along before the old media went bad and I copied it all } over. One thing I don't like about CD-ROMs is they are not protected from } scratching or being broken. } } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 }
On the HP CD Writer there are two programs for writing to a CD.. Out here we use the CD-R one time write to the disc.
There is the HP program for writing a CD which does not have a way to verify the CD was written. There is also another program that comes with the CD writer from HP that performs a write speed test on the disc then writes to the disc and then it verifies the disc... the program is called MY CD on the HP CD writers....
How may I calculate (OK, make an educated guess) the temperture on my grid in the TEM? I know it depends on the high voltage, the beam current, the thickness of the specimen, the contact between grid and holder, and the phase of the moon, and so probably I won't *really* know. All I need is a ballpark figure, like is it 80C or 200C or 1500C?
I'm watching something presumably melt and fuse/crystallize on my LEO 912 EFTEM at 100-120 kV with a beam current of somewhere in the neighborhood of 10 uA. This is not necessarily a bad thing - it tells me something useful about these Si nanoparticles!
Mahalo, Tina
Sunny, clear, about 74F, South Shore flat, North Shore up.
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
How may I calculate (OK, make an educated guess) the temperture on my grid in the TEM? I know it depends on the high voltage, the beam current, the thickness of the specimen, the contact between grid and holder, and the phase of the moon, and so probably I won't *really* know. All I need is a ballpark figure, like is it 80C or 200C or 1500C?
I'm watching something presumably melt and fuse/crystallize on my LEO 912 EFTEM at 100-120 kV with a beam current of somewhere in the neighborhood of 10 uA. This is not necessarily a bad thing - it tells me something useful about these Si nanoparticles!
Dear Tina, I posted something similar to the list a while ago, and wrote up a more complete version for Microscopy Today. Although the variables are different, the technique should be the same. A number you'd need would be the stopping power of Si for 100 kV e-, which is 3.274 Mev cm^2/gm. The radiative stopping power is much smaller than the collisional stopping power, which is 3.265 MeV cm^2/gm (since the radiation should escape the particle, use this latter number). Use this to calculate how much energy is transmitted to the specimen for each electron by multiplying the stopping power by the density of Si (2.33 gm/cm^3) to get how much energy is deposited per unit path length, figuring out the path length of each electron, and using the beam current flux (in e-/nm^2 or the like) to get how many e- strike the particle each second. Next, calculate the heat losses by conduction and radiation as a function of temperature, and the steady-state temperature of the specimen--the number you want--will be where the heat absorbed is equal to the heat lost. I assumed that heat radiated and that conducted from the immediate vicinity of the particle would be lost in an essentially infinite heat sink, and that the moon was in third quarter.
Sunny, clear, about 74F, South Shore flat, North Shore up.
Cloudy, about 270 K, Hudson icy. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Two minerals are fairly easy to come by Zircon (SiZrO4) and Hafnon (SiHfO4) or could be synthesized. The rare-earth elements more commonly form phosphate minerals such as monazite ((Light REE)PO4) and xenotime ((Heavy REE,Y)PO4). The substitution of Huttonite (SiThO4) into these structures would allow for the calculation of REE,Y/Si k-factors. The problem with these minerals is that they are normally not homogeneous, so special care must be made to correlate electron microprobe with TEM analyses. Alternatively, you could try to synthesize your own standards. Hope this helps, Ken } } Hi, everybody ! } } I would greatly appreciate some hints concerning the kind of materials } (minerals ?) that } could be used in order to determine the k(A,B) (Cliff-Lorimer approx.) } factors for my instrument } (TEM CM-200 Philips microscope with EDAX EDS X-ray spectrometer) concerning } Zr(K-lines), } Y(K-lines), Hf(L-lines) and La(?-lines). Do they exist Si containing minerals } which could be } used to determine the k factors relating Si and the above mentioned elements ? } Are they suppliers of standard materials that could be used for determining } the k factors for the } above mentioned elements ? } } Thank you in advance ! } } Corneliu Sarbu, PhD } Dept.of Metallurgy and Applied Materials Science } Catholic University of Leuven } Belgium
Around 20 years ago, I saw a movie about killer bats in which, after looking at a slide with what looked like a Tasco microscope, the scientist exclaimed "It's just as I feared - the rabies bacillus!". To add insult to injury there followed a view through the microscope where, if I recall correctly, a paramecium was swimming.
It's my understanding that dye-based CD-ROMs aren't archival, but the gold-based version is. Anyone know more about this?
Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
sorry for bothering you with that. I wanted to send an email to Phil Oshel, but deleted the email before writing down his email address. Phil, please contact me off-list, thanks.
Andreas
************************************************* Dr. Andreas Taubert Materials Science and Engineering Dept. 3231 Walnut Street The University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
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Reply to: Re: Ultimate and perhaps last of TV Trivia Dear All,
I too have been enjoying this conversation and thought I would sit back and read. However, I can't resist a few comments. After all we have one of the actual Zeiss microscopes from Jurassic Park in our lab. It works wonderfully, as a Zeiss microscope should. The only madification they made was to paint it pink to look right on film. Being so close to Hollywood, we have a few other pieces of famous furniture around the lab. I must admit that I too look out for appearances of microscopes, and especially EM's in the media, but I think I must be a little more tolerant of the way we are portrayed. My phylosophy is that as long as we are out there, then it is a good thing. Who cares if the portrayal is accurate or not, we can sort that out in our lectures. At least we are being recognized. After all, it is much better to be talked about (no matter the subject) than to be ignored.
If we systematically went through film portrayals of any subject then I am sure we will find Hollywood had managed to insult just about every scientific disipline, ethnic group and foreign country. Who cares, its only entertainment. Watch out instead for the school science books that show cells with organelles but do not show the Golgi complex, or the histology books that omit the lysosomes. They are there. This is far more harmful to the scientific community.
I enjoyed the comments from Roger Moretz about the speed samples are processed. After all one of the roles of this form of entertainment has become a predictor of the future. Maybe the portrayals are so so inaccurate after all. We already work with microscopes in full daylight (from computer screens), we are close to having 4hr sample processing for resin-embedded samples (cf microwave processing), and we can already section and examine biopsies in less than 2 hr (with cryosectioning methods). Two thumbs up for Hollywood for showing us the way.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Roger Moretz wrote:
} Or not. I guess I just can't ignore this thread. I have always been vocal } about inconsistencies/stupidities/errors in TV/movies/etc. So, as to } Andromeda Strain: first of all, there was the insertion of the specimen } without use of an airlock (and those of us who suffered with the Forgflo/nee } RCA EMU-4 know that the beast had this horrid airlock that used the external } bellows minipump!!!); then there was the immediate location of the desired } area under the beam....; plus all those already mentioned. On Quincy: the } first episode (or maybe 2) the lab was fully outfitted with Zeiss equipment; } the rest of the series the lab was outfitted with AO (must be nice to have } that kind of equipment budget--but why go that direction??? could it be } politics???); then the SEM/microprobe episode, where, once again, the area } of interest with the decisive inclusion was magically right in the area } being examined immediately; then there was the TEM episode where Sam } received the biopsy about 4am and had a block, stained sections and a } confirmatory diagnosis by 8am (now there's a reality check for you--did your } boss pick up on that and demand that turn-around for you????); I'm sure } there were others but those stuck (mostly in my craw). And a final overall } plaint about the fact that everybody was working (at high mag, no less) in } brightly lit rooms, when we toiled away in near pitch dark!! Even that } Forgflo/RCA with its ma beam current couldn't do that--I know from many } hours in the dark, dark adjusting my eyes. } } O well, as someone put it--that's Hollyweird. } } Roger Moretz Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals, Inc. } On Sat, 06 Jan 2001 11:19:43 -0800, Robert Ruscica wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I have enjoyed reading all the TV trivia regarding the Quincy and } Andromeda } } Strain. What many of you may not know is that for the "Strain" movie the } } makers of the film contacted a now defunct Microprobe Company, Materials } } Analysis Co. We sent to the studio an Electron Microprobe. The only } portion } } that made the film was some blinking x-ray scalers and a portion of the } } electronics rack. What the impact of microprobe was in the film I can't } } guess or remember. } } In the Quincy episodes, there were more than one, we sent to the studio } an } } ISI SEM along with a service engineer, whose name at the moment escapes } } me, to install it and our application and Demonstration man, Bill Roth } to } } run the SEM, Bill has been with Hitachi for many years now and could give } } } more incite as to what happened at the studio than I can as he was there } } for a week doing the one episode. I remember him telling me that Quincy's } } } technician, Sam was explaining to Quincy what was on the CRT with the } } camera going from the control panel with Bill's hands being filmed but } the } } overall shot of Sam and Quincy looking at the CRT and discussing the } } forensic material. I still have some 8x10's around here somewhere of the } } filming. Thought you might be interested. } } } } Regards, } } Bob Ruscica } } } } } } } } } } _______________________________________________________ } Send a cool gift with your E-Card } http://www.bluemountain.com/giftcenter/ } } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id AC703E6C025E; Mon, 08 Jan 2001 16:33:52 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id JAA13624 } for dist-Microscopy; Mon, 8 Jan 2001 09:16:16 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id JAA13616 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 8 Jan 2001 } 09:15:46 -0600 (CST) } Received: from ewey.excite.com (ewey-rwcmta.excite.com [198.3.99.191]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id JAA13608 } for {Microscopy-at-sparc5.microscopy.com} ; Mon, 8 Jan 2001 09:15:34 -0600 (CST) } Received: from spike.excite.com ([199.172.152.97]) by ewey.excite.com } (InterMail vM.4.01.02.39 201-229-119-122) with ESMTP } id {20010108151408.VOMO24504.ewey.excite.com-at-spike.excite.com} ; } Mon, 8 Jan 2001 07:14:08 -0800 } Message-ID: {18272603.978966848378.JavaMail.imail-at-spike.excite.com} } Date: Mon, 8 Jan 2001 07:14:07 -0800 (PST) } From: Roger Moretz {rcmoretz-at-excite.com} } To: Robert Ruscica {ruscica-at-etp-usa.com} , Microscopy-at-sparc5.microscopy.com } Subject: Re: Ultimate and perhaps last of TV Trivia } Mime-Version: 1.0 } Content-Type: text/plain; charset=us-ascii } Content-Transfer-Encoding: 7bit } X-Mailer: Excite Inbox } X-Sender-Ip: 63.80.54.82 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273278772 } Status: U }
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They are all dye-based. What is silver or gold is the coating on the top of the media. It ca readily be flaked off.
With the metal coating removed, the media is generally transparent. Some have a blue tint while others have little color. Others might look green. But essentially, they are all chalgocencide technology. Some are better than others.
gg
At 03:49 PM 1/8/01, you wrote:
} Colleagues, } } It's my understanding that dye-based CD-ROMs aren't archival, but the } gold-based version is. Anyone know more about this? } } Dee } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 914/365-8640 } F: 914/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995)
These are found all of the time in Hollywood. They are stand-ins. Only a very sharp eye can tell the difference. Notice how well Antonio Banderas dances in "The Mask of Zorro?" Real, or a stand-in? duh.
gary
At 03:49 PM 1/8/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Tina, I'm sure this question has been asked before, (look in the archives) - but I think the basic answer is 'it depends'(!). I am currently having terrible trouble with plucked FIB sections of Au metallised GaAs on holey carbon films; the metal melts, alloys with and destroys my specimen even at relatively low beam currents in my JEOL 120 CX. I have been told by our theory guys that little heat will escape by radiation below about 500C, so if you have a sample with very poor thermal conductivity it will be easy to get to this temperature. You may be able to avoid the problem somewhat by using higher accelerating voltages (less inelastic interaction with the specimen, I believe) and/or adding a carbon coating to the specimen to increase thermal conductivity.
Good luck!
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Happy New Year to you all! } } How may I calculate (OK, make an educated guess) the temperture on my grid } in the TEM? I know it depends on the high voltage, the beam current, the } thickness of the specimen, the contact between grid and holder, and the } phase of the moon, and so probably I won't *really* know. All I need is a } ballpark figure, like is it 80C or 200C or 1500C? } } I'm watching something presumably melt and fuse/crystallize on my LEO 912 } EFTEM at 100-120 kV with a beam current of somewhere in the neighborhood } of 10 uA. This is not necessarily a bad thing - it tells me something } useful about these Si nanoparticles! } } Mahalo, } Tina } } Sunny, clear, about 74F, South Shore flat, North Shore up. } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
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It's a bit late but may I wish you a happy new year.
My personal experience is that the biggest problem with CD writing is the possibility of corrupting the disk and not noticing (e.g. buffer under runs). This can at the least result in wasted time and frustration and at worst the actual loss of data if not properly backed up elsewhere. I haven't seen this mentioned so I thought I would bring up the subject of 'Burn proof' CD writers which have appeared in the last 6-12 months. They apparently have built in hardware/software which should mean that you can now use CDs as super floppies and not have to worry about turning off conflicting software before writing large chunks of data to CD. The magazine reviews have raved about this stating that you could play 'Quake' or access the internet and still write to a CD without error. Has anyone had any experience of these new drives?
Archival quality of CDs, ZIPs and magneto opticals has been discussed many times before and the usual conclusion is that the technology will be defunct before good quality discs, stored properly will be significantly corrupted. My money has to stay with CDs as a cheap, reliable and 'likely-to-be-around-in-a-decade' technology (e.g. all modern DVD drives still read CDs). CD/DVD technologies still have a long way to go in terms of data density and so multi-format reading machines would seem more likely than with magnetic media ... unless you know better. I'm sure that I read of recent developments in dyes/pigments (maybe it was Kodak - I don't know) that should greatly reduce writeable CDs deterioration in light. One thing's for certain if I want to distribute images to students, researchers or other users there is only one universal technology (some new PCs don't even have a floppy drive now).
Another thread was discussing e.m. film scanners and I think I should mention the Epson Perfection 1200P which is a USB scanner that can be used with A4 prints or 5x4 inch negatives. It has a true resolution of 1200dpi and an optical density range of 3.2 and costs a little over 200 UK pounds. Much of our work with undergraduates and researchers has been done at 600dpi (which should easily allow an equivalent of 3x print enlargement) and dare-I-say-it JPEG formats (set at lowest compression to retain most detail). The film is, after all, our archive and the digital images are at present for cataloguing, rapid retrieval and project write ups - the students love it if only because they don't spend hours in the darkroom. I am not suggesting that this particular scanner will suit all users because it has a narrower OD range than some, but is an excellent low cost way of trying the technology out while it is still maturing. The biggest problem is that there was no film holder for our format but cardboard and sticky tape have solved that problem, temporarily. Also if you opt for the USB version then you really need Windows 98 on a PC (95 - including the the second version, NT etc. may all have problems).
I have no commercial interest in any of the above technologies unless they make my work or research any easier - in a year or two I may be in a better position to judge that, objectively.
Malcolm Haswell e.m. unit University of Sunderland UK
ERIC wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } On the HP CD Writer there are two programs for writing to a CD.. Out here } we use the CD-R one time write to the disc. } } There is the HP program for writing a CD which does not have a way to } verify the CD was written. There is also another program that comes with } the CD writer from HP that performs a write speed test on the disc then } writes to the disc and then it verifies the disc... the program is called } MY CD on the HP CD writers....
I agree, but I don't blame the film makers. They are part of the majority of the public who (sometimes proudly) proclaim to have no/little knowledge of science. It seems to be quite common to use bugs, germs, viruses, bacilli and bacteria as interchangeable terms. This can be extremely misleading when a national news reporter warns of the latest outbreak of the meningitis 'virus' when he almost certainly meant the bacterial form.
There has been more than one occasion where I have seen a TEM instrument with a superb SEM image portrayed on film or TV (yes I know about TEM/SEM/STEMs - but it wasn't) presumably because the TEM looks impressive and so do SEM pictures. I also saw the Bladerunner SEM and another technology it had was a voice activated TV with high resolution scanner and printer. It seemed well out of reach of the average household at the time and although I haven't seen this on a TV this month all of that technology is common on your average home PC now - maybe the next Intel (QX4) microscope will be a toy SEM connected to your TV.
Malcolm Haswell e.m. unit University of Sunderland UK
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Around 20 years ago, I saw a movie about killer bats in which, after looking } at a slide with what looked like a Tasco microscope, the scientist exclaimed } "It's just as I feared - the rabies bacillus!". To add insult to injury there } followed a view through the microscope where, if I recall correctly, a } paramecium was swimming. } } Paul
At 07:22 PM 1/8/01 -0800, Gary Gaugler wrote: } They are all dye-based. What is silver or gold is the coating } on the top of the media. It ca readily be flaked off.
No, supposedly the gold ones are actually made of gold the metal, and the "silver" ones are aluminum. The Al will oxidize over time if scratched, the Au won't.
The story says that if you scratch a disc deep enough to expose the metal, it could degrade the data over time. I don't put a lot of stock in this worry, though. After all, it wouldn't take much more of a scratch to wipe out a section of gold, too, and there's a big difference in damage between a radial and a concentric scratch on any media.
Thanks for coming up with the reference. I just KNEW it was it my Reference Manager database (NOT). So, for several days now I have been trying to figure out if I ever referenced it in a paper, or in notes or methods/techniques/procedures.... Now I can sleep at night again :)
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Mon, 08 Jan 2001 12:45:29 -0800, Michael Nesson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Gordon Couger wrote: } } } } } Could someone post the reference for the article for making glass knives } } by hand? } } Here's the original reference, which includes a bit of the history of Ralph } knives (in honor of the late Dr. Paul Ralph): } Stain Technology 51(2): 71-97. [1976]. Bennet et al. } Science and art in preparing tissues embedded in plastic for } light microscopy, } with special reference to glycol methacrylate, glass knives and } simple stains. } } Good luck. } Mike Nesson } _______________________________________________________________________ } Michael Nesson, Ph.D. Department of Biochemistry & Biophysics } 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 } (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu } } }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Our customers have enjoyed success with the Agfa "Duoscan" line of scanners. Fitting in your price range would be the Duoscan Hi-D. The Hi-D has a Dmax of 3.8 (hence the name Hi-D), 1000x2000 hardware resolution, apochromatic optics, excellent software for both Mac and Windows, color management software, and a SCSI interface. The Hi-D sells for under $2600. Coming soon will be the Arcus 1200, which also has glassless film scanning, 1200x2400 hardware resolution with a SCSI interface. The Arcus 1200 will be approximately $800-850 when available. The Agfa T2500 is a similar design, with 2500x2500 hardware resolution and 3.4 Dmax. The T2500 does not fall in this price range, selling for under $4300. It is a phenomenal scanner for TEM. There is currently a $350 rebate on the T2500. All of these scanners share the Agfa Fotolook sofware driver for Mac and Windows. Unlike most "flatbed" scanners which place films on a glass bed for scanning, the Duoscan line utilizes a glassless holder similar to a negative carrier in an enlarger. There is no glass between the film and the CCD. This eliminates Newton Rings and other issues associated with scanning through glass.
Microtek International shares some hardware specs with the Agfa Duoscan line in their Artixscan series. The Artixscan 1100 would be the "sister" model to the Duoscan Hi-D with 1000x2000 optical resolution, 3.9 Dmax, SCSI interface,etc. Price is under $1750. The Artixscan 2500 is similar to the Duoscan T2500- 2500x2500 optical res, SCSI interface, etc. Price under $4000.
See Microtek at http://www.microtekusa.com/list.zhtml?cid=2 Agfa DuoScan HI-D at http://www.agfahome.com/product/CatProd_DisplayPublic.html?id=4290 Agfa DuoScan T2500 at http://www.agfahome.com/product/CatProd_DisplayPublic.html?id=3115 Agfa Arcus 1200 at http://www.agfahome.com/product/CatProd_DisplayPublic.html?id=6130
I will be happy to provide literature, information or answer any questions. Please contact me directly by phone or email.
Sincerely,
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
Hello, I was wondering if any one might know where I could get some cryostat tissue mounting stubs. I am looking for the 'T' type looking stubs that fit into a ball joint so the stub position can be adjusted in the cryostat chuck. The ones I have used were made of copper and the pin diameter was1/8" with a locking screw in the shaft of the ball joint. Thanks for any leads. Sincerely, Jonathan
Jonathan Wilson (Ph.D.) Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR) Universidade do Porto Rua do Campo Alegre 823 4150-180 Porto, Portugal tel 351 22 606 0421 fax351 22 606 0423 e-mail: wilson_jm-at-cimar.org mop01258-at-mail.telepac.pt
Many years ago (at least 6!) when we had an early Pinnacle Micro CD writer I got into the habit of checking my CD's by copying them back to the hard drive. If they would copy without generating an error then they were OK (an even better way was to do the copy on another CD drive). One fairly often found one that was bad.
Today, using a more modern drive and faster computer I still do the same thing (my software doesn't have a verify option), but I rarely find a bad disk. However, it doesn't take much of my time, and I always think better safe than sorry when it comes to an archive!
I need to get some parts for a RMC MX-600XL ultramicrotome.. I know RMC was sold to another company from Ventana.. How do I or who do I contact about some parts??
Eric A. Rosen UCLA Medical Center Dept. Pathology and Lab Medicine Los Angeles, CA 90095
In the context of recent queries, I just thought I'd post Nikon's most recent announcement ... see: http://www.klt.co.jp/Nikon/Press_Release/ls-8000.html
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
In the context of recent queries, I just thought I'd post Nikon's most recent announcement ... see: http://www.klt.co.jp/Nikon/Press_Release/ls-8000.html
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
In a message dated 01/09/2001 11:30:40 AM US Mountain Standard Time, biology-at-ucla.edu writes:
{ { I need to get some parts for a RMC MX-600XL ultramicrotome.. I know RMC was sold to another company from Ventana.. How do I or who do I contact about some parts??
Eric A. Rosen UCLA Medical Center Dept. Pathology and Lab Medicine Los Angeles, CA 90095
} }
Eric,
RMC is now a part of Boeckler Instruments in Tucson, Arizona. You can reach them at:
Boeckler Instruments, Inc. 4650 South Butterfield Drive Tucson, AZ 85714 Tel. 800-552-2262 Fax 520-745-0004
I have a Kevex EDS detector that was shipped via FEDEX with liquid nitrogen. FEDEX, in their infinite wisdom, managed to tilt the detector and drain the LN2 from the dewar. (Although the packing box and "tilt watch indicators" say "do not tilt', "this side up", etc.)
Upon inspecting the detector I have found that a plug on the side of the dewar has been removed and the detector is at atmosphere. I suspect that the LN2 froze the plug & "O" rings therby allowing air into the detector.
I am considering machining an adapter flange and repumping the detector. This is, of course, assuming that the detector window is entact.
Does anyone have experience with this? While pumping, I am considering pouring hot water into the dewar to outgass the system but I don't know if this is a good idea.
Hi - I am looking for a used lens regulator circuit board for a Philips TEM 201. The parts # is Lens Regulator 5322 695 14022 in the 1974 service manual -also called RB-U13. Is anyone junking a 201? The price from FEI/Philips is a bit beyond my budget! g
Hi - I am looking for a used lens regulator circuit board for a Philips TEM 201. The parts # is Lens Regulator 5322 695 14022 in the 1974 service manual -also called RB-U13. Is anyone junking a 201? g
Colleagues, Has anyone had any experience with generating energy filtered diffraction patterns (not CBED) with a Gatan GIF on a Philips CM? I would like some pointers on intensity issues and summation of images. Thanks in advance. Ciao for now, Ken
We are installing a new JEOLCO JSM 6700F Field Emission Scanning Electron Microscope. This is a new model and incorporates some novel technology. We are interested in communicating with others who are or will be working with this model. Please respond off-line and I will construct a mailing list to distribute questions, concerns and, of course, suggestions for operation and improvement.
James R. Coleman, Ph.D. Supervisor, Electron Microscopy Laboratories Boehringer-Ingelheim Pharmaceuticals Inc.
Richard asked if the exposure meter on the Nikion coolpix 900 digital camera was sensitive enough to control exposure in low light (immunofluorescent) conditions. We used the camera to take images of samples labeled with moderate intensely with FITC. We captured nice images in the "spot" automatic metering mode. With the camera mounted in one ocular, the camera chose to expose at F 3.1 and kept the shutter open one second. However, it was a bit difficult to use since the image did not appear on the display during focusing. There was not enough light for real time imaging. To insure a focused image, we needed focus in brightfield illumination, then go to EPI to collect the image. You could also use a monitor, but that would add significant cost to the system.
It may be useful to realize that the camera has the ability to be used in several automatic exposure settings (programmed, aperture priority, shutter priority) (wide field, narrow field, center spot) and in manual mode, where you can use a timed exposure from 1/1000 to 8 seconds, or longer (up to 60 seconds) by pushing the exposure once to open and again to close.
Feel free to phone with more questions,
Doug
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
It all depends. If there is no-one else who would pay to fix your detector, then you have nothing to lose by doing what you suggest (incl. the hot water). I do it relatively often, (but only to detectors that are seriously degraded or not functioning at all), and it works, but I've never had a crystal return to tip-top premium performance after this treatment. (In fact, only today I deliberately vented a detector and re-pumped it, but that is a long, and different, story). BTW, you only need to achieve a quite modest vacuum during the pump - the molecular sieve will take care of a lot of gas when it cools! Having said that, I have usually rigged up a piece of vacuum hose from the pumping adapter to a port I had added to an old evaporater, and pumped down to the 10^-5 - 10^-4 range. I have a pumping adapter with a US thread for the plug - perhaps it will work on your Kevex detector (in fact, it came from Kevex 20 years ago). If you want to borrow it, let me have your FedEx or other shipping account number, and shipping details, and I'll send it off.
On the other hand, if the shipper, or FEDEX, can be persuaded to pay to have the detector refurbished, it will probably work better in the end, and a do-it-yourself attempt beforehand may make your claim more difficult!
Good luck!
Tony
Tony.
At 02:07 PM 1/9/2001 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I suppose, if plastic film or plastic sections (biological samples) are relatively stable under the beam, it may mean (may not) that temperature is not so high. In my hands, overheating of the plastic film in air over +150oC will destroy the film. I assume, something like that would happens in the microscope if the beam generate such amount of heat (plastic film itself has a low thermal conductivity, I believe). Another scenario: unelastic scattered electrons may transfer kinetic energy (not necessary it will immediately transferred into the heat) to the sample/film. This excess of energy may heat the sample but may force molecules/atoms to migrate for instance. Heavy atoms should affected more. I do remember, 20 or so years ago people shout films under TEM, how Pt or Au atoms may move under the beam. They (atoms) tend to form big clusters under the beam. This is explanation (to me), why the resolution of Pt or Au shadowed samples is so bad. Pt-C shadowing succeeded because carbon protect Pt from migration and forming big clusters. In general, I do agree: "it depends"...
Sergey
At 09:39 AM 1/9/01 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I did a few detector treatments and I agree with Tony's comments. One addition: Obtain a fairly good vacuum before pouring boiling water (couple of liters) into the dewar. The heat causes a dramatic drop in vacuum due to outgasing of the molecular sieve. I would terminate pumping a couple of hours after the hot water was added. Obviously is nice to have a clean vacuum system available as backstreaming oil vapour would rather defeat the purpose. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, January 10, 2001 1:37 PM, Tony Garratt-Reed [SMTP:tonygr-at-mit.edu] wrote: } } } Earl- } } It all depends. If there is no-one else who would pay to fix your } detector, then you have nothing to lose by doing what you suggest (incl. } the hot water). I do it relatively often, (but only to detectors that are } seriously degraded or not functioning at all), and it works, but I've never } had a crystal return to tip-top premium performance after this treatment. } (In fact, only today I deliberately vented a detector and re-pumped it, but } that is a long, and different, story). BTW, you only need to achieve a } quite modest vacuum during the pump - the molecular sieve will take care of } a lot of gas when it cools! Having said that, I have usually rigged up a } piece of vacuum hose from the pumping adapter to a port I had added to an } old evaporater, and pumped down to the 10^-5 - 10^-4 range. I have a } pumping adapter with a US thread for the plug - perhaps it will work on } your Kevex detector (in fact, it came from Kevex 20 years ago). If you } want to borrow it, let me have your FedEx or other shipping account number, } and shipping details, and I'll send it off. } } On the other hand, if the shipper, or FEDEX, can be persuaded to pay to } have the detector refurbished, it will probably work better in the end, and } a do-it-yourself attempt beforehand may make your claim more difficult! } } Good luck! } } Tony } } Tony. } } } At 02:07 PM 1/9/2001 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have a Kevex EDS detector that was shipped via FEDEX with liquid nitrogen. } } FEDEX, in their infinite wisdom, managed to tilt the detector and drain the } } LN2 from the dewar. } } (Although the packing box and "tilt watch indicators" say "do not tilt', } } "this side up", etc.) } } } } Upon inspecting the detector I have found that a plug on the side of the } } dewar has been removed and the detector is at atmosphere. I suspect that the } } LN2 froze the plug & "O" rings therby allowing air into the detector. } } } } I am considering machining an adapter flange and repumping the detector. } } This is, of course, assuming that the detector window is entact. } } } } Does anyone have experience with this? While pumping, I am considering } } pouring hot water into the dewar to outgass the system but I don't know if } } this is a good idea. } } } } Thank You, } } } } Earl Weltmer } } } *********************************************** } Anthony J. Garratt-Reed, M.A., D.Phil. } Principal Research Scientist } Room 13-1027 } Massachusetts Institute of Technology } 77 Massachusetts Avenue } Cambridge, MA 02137-4307 } } Tel: (617) 253-4622 } Fax: (617) 258-6478 } ***********************************************
If you have a problem locating the original suppliers of these object holders, I should be able to make them for you, please fax me a sketch will measurement and I will reply with a price.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England
Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: AlanBright-at-brightinstruments.com Web Site: www.brightinstruments.com
-----Original Message----- } From: Jonathan Wilson [mailto:wilson_jm-at-cimar.org] Sent: 10 January 2001 00:31 To: Microscopy
Hello, I was wondering if any one might know where I could get some cryostat tissue mounting stubs. I am looking for the 'T' type looking stubs that fit into a ball joint so the stub position can be adjusted in the cryostat chuck. The ones I have used were made of copper and the pin diameter was1/8" with a locking screw in the shaft of the ball joint. Thanks for any leads. Sincerely, Jonathan
Jonathan Wilson (Ph.D.) Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR) Universidade do Porto Rua do Campo Alegre 823 4150-180 Porto, Portugal tel 351 22 606 0421 fax351 22 606 0423 e-mail: wilson_jm-at-cimar.org mop01258-at-mail.telepac.pt
For about 15 years, I ran a Kevex "extra" detector. This was a windowless type and over time (I did a lot of light element work) the crystal would ice over. The Etec was turbo pumped, but between specimen out gassing and the methane from the WDS, icing over time could not be avoided. I warmed and pumped a number of times using warm/hot water. A small amount of degradation occurred, but it did help overall. I later went to a hot air gun and a tube to direct the air to the bottom of the dewar. By doing this, I did not have to worry about drying the dewar.
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Several folks have asked about chillers and evaporators lately.
We have a Denton DV-515 vacuum evaporator which has been replaced by an automated DV-502A. The DV-515 was originally from a government lab so it has been well maintained and cared for. We had used it mainly as a back up for our DV-502A. We use the carbon rod arc to produce TEM support films and general carbon coating of samples on MCE & PC filters during TEM specimen prep and to put carbon coatings on SEM samples.
If you are interested, we would like 2500. reply to:
All, The preliminary information I have is that the Coolscan 8000 ED will sell for approximately $4000 US. We will carry it when it becomes available, currently listed as June. I hope to see it in early February. Quick specs.. 4000dpi optical resolution IEEE1394 "Firewire" Interface- not SCSI 4.2 dynamic range 48bit images Multi Sample Scanning
Info is also at http://64.77.39.20/usa_product/product.jsp?cat=7&grp=703&productNr=9246
I'll pass along more info as it becomes available.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com } } This looked pretty interesting. Have you heard a ballpark price? } } } } } In the context of recent queries, I just thought I'd } } post Nikon's most recent announcement ... see: } } http://www.klt.co.jp/Nikon/Press_Release/ls-8000.html } }
I called Nikon yesterday (1/9/2001) to ask about the Coolscan 8000 ED. They told me it would cost $3000 (U.S.) and is scheduled to be released in April.
Question: I need to view calcium carbonate crystals with a transmission electron microscope for a research project. I also need to record the images. All I've managed to do so far is order thin carbon grids. I have been asked to write a plan for my experiment but I have no clue how to prepare crystals in solution for magnification or what tools to use. Please give me some direction. My professors say I should figure everything out myself - "It will be a good learning experience."
Dear Jane, The most difficult aspect of this is likely to be getting sufficiently small crystals dispersed over the grid surface so that you can see the individual crystals. You do not say whether you need detail within the crystals or whether just seeing silhouettes is sufficient. If the latter, then just evaporate the solvent rapidly from a dilute CaCO3 solution placed on the grid. This should leave small crystals dispersed over the grid. If you can't evaporate on the grid, you will have to prepare the small crystals and transfer them either by suspending them in a volatile liquid which doesn't dissolve CaCO3 or by wafting them into the air and collecting them on the grid. If you need detail within the crystals, they must be very thin--no more than ~10 nm (depending on the EM voltage), so you will have to find a way to get them this thin, and, if possible, much wider than they are thin--a platy habit. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Email: us004118-at-mindspring.com Name: Leonard Lessin, FBPA
School: (Retired Science & Medical Photographer)
State: NY
Zip: 10012
Question: I am enjoying doing photomicrographs of crystal preperations in polarized light.However I have insuffient knowledge of chemistry to choose solvents without a long series of trial and error efforts.Can you give me a rationale and/or a reference to go about this in a more productive manner?
Dear Leonard, There are several requirements for a suitable solvent: 1) it must, of course, disolve the material you want to examine, 2) it must evaporate to produce crystals in a habit suitable for your work (not too small, transparent, etc.), and 3) it should not be too toxic even though you may have a fume hood available. The key to 1) is "like disolves like". If you have an ionic salt or a polar solute such as sucrose, water would be a good choice, if you have an aromatic compound, such as naphthalene, an aromatic would work (I hesitate to reccommend benzene, even though it gives nice crystals, because of its toxicity; I would try toluene.), and if you have a lipid, like cholesterol, I'd use a non-polar aliphatic solvent such as hexane. For 2) it is important to have a suitable rate of evaporation, so in the last case, hexane might evaporate too fast, so I would go to a longer-chain hydrocarbon, e.g., decane. The Handbook of Chemistry and Physics lists many compounds, both organic and inorganic, and some solvents for each; I'd use that as a first reference. I hope this will reduce the problem from a long series of trial-and-error efforts to a short one, but you can still expect some problems. The preparation of suitable crystals is most often the most difficult part of these studies. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
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I was forced to dismantle my Opti-quip fluorescence bulb housing the other day for an unrelated problem. The 75W xenon bulb only has 200 hrs of use so I would have expected it to have another 200 hrs of life based on what the supplier told me. But I noticed that at one of the ends, at the junction between the metal cap and the glass bulb, there was a buildup of a hard, white substance on the outside of the glass. Does any one know what caused this and if it is safe to re-install the bulb? TIA, tom
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Thermo NORAN, the makers of the finest instrumentation for elemental analysis is seeking both a Microanalysis and an XRF Applications/Product Specialist. This position will be responsible for the ensuring the success of the applications/products targeted at analytical instrument markets, including materials characterization and materials development markets. Will perform demonstrations and provide other pre-sale support as required at major trade shows, technical conferences and other meetings. Will analyze customer samples and prepare reports for prospective customers. Worldwide travel may be required to perform demonstrations and presentations. Travel may include trips greater in duration than one week. Must be degreed in a physical or life science, have excellent written and oral communication skills. Knowledge of electron microscope and/or x-ray microanalysis with experience in materials characterization. Interested candidates should send, e-mail or fax resumes to:
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I'm writing up a wish list for our EM lab, and it includes (gasp) light microscopes. My question is - how do I go about evaluating and choosing a digital camera for light microscopes? It would be for both compound and dissecting microscopes, should be color, decent resolution, not necessarily low light nor real-time video, but capable of good images for image analysis on sections. We are getting a confocal, so fluorescence imaging would be done there rather than with the proposed 'scope and camera.
What do I need to look for, and what price ranges are we talking about?
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Does anyone know how to mount large (100um+) diatoms for LM or SEM? The little ones are OK, they stick to most any stub, but these big guys like to fall off.
Anyone know how arranged diatom slides are/were made? I checked WWW but no specifics on techniques. Something like these methods would work for us.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Richard asked if the exposure meter on the Nikion coolpix 900 digital camera was sensitive enough to control exposure in low light (immunofluorescent) conditions. We used the camera to take images of samples labeled with moderate intensely with FITC. We captured nice images in the "spot" automatic metering mode. With the camera mounted in one ocular, the camera chose to expose at F 3.1 and kept the shutter open one second. However, it was a bit difficult to use since the image did not appear on the display during focusing. There was not enough light for real time imaging. To insure a focused image, we needed focus in brightfield illumination, then go to EPI to collect the image. You could also use a monitor, but that would add significant cost to the system.
It may be useful to realize that the camera has the ability to be used in several automatic exposure settings (programmed, aperture priority, shutter priority) (wide field, narrow field, center spot) and in manual mode, where you can use a timed exposure from 1/1000 to 8 seconds, or longer (up to 60 seconds) by pushing the exposure once to open and again to close.
Feel free to phone with more questions,
Doug
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Listers My company's safety folks are bent on making us wear Kevlar gloves while razor trimming samples for ultramicrotomy - I'm talking about the final trimming done using a trimming stand on the microtome. As you might know, this is fine work - wearing gloves is a handicap. My questions: 1. Does anyone out there actually wear gloves? - if so then can you recommend a glove? 2. Are there other ways to make this task look less dangerous to the safety police? (short of purchasing a trimming machine)
Thanks!
Dave Calvert Eastman Chemical Company Building 150B Lincoln Street (packages) P.O. Box 1972 (US Mail) Kingsport, Tennessee 37662-1972 (423) 229-4943 (423) 229-4558 fax
} My company's safety folks are bent on making us wear Kevlar gloves while } razor trimming samples for ultramicrotomy - I'm talking about the final } trimming done using a trimming stand on the microtome. As you might know, } this is fine work - wearing gloves is a handicap. } My questions: } 1. Does anyone out there actually wear gloves? - if so then can you } recommend a glove? } 2. Are there other ways to make this task look less dangerous to the } safety police? (short of purchasing a trimming machine)
When I train people to trim blocks, I always point to the Band-Aids first, then to the razor blades! This gets a laugh, but drives home the point. Then I suggest that they put the bandages on FIRST before trimming, which also gets a laugh, but significant number of people decide to do so! Therefore, I suggest something like finger cots or other wrap-around solutions that leave some dexterity while protecting the last joints of the fingers.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Jonathan Krupp wrote: ============================================================== Does anyone know how to mount large (100um+) diatoms for LM or SEM? The little ones are OK, they stick to most any stub, but these big guys like to fall off.
Anyone know how arranged diatom slides are/were made? I checked WWW but no specifics on techniques. Something like these methods would work for us. ============================================================== Consider these two options:
a) Tacky Dot™ Slides, see URL http://www.2spi.com/new/tacky.html and b) Double sided conductive discs, sheets or tape http://www.2spi.com/catalog/spec_prep/cond_adhes.html
The Tacky Dot Slides have good adhesion and the diatoms should be held in place without problems. The double sided conductive sheets, discs, and tape should work as well, but if you are using material that has aged past its "expiration date", the surface tends to lose its "tac" and indeed might not hold the "big guys". Of course, only the Tacky Dot Slides will result in the diatoms being arranged in orthogonal arrays for convenient analysis.
Disclaimer: SPI Supplies is the exclusive worldwide licensee for the manufacturing and distribution of Tacky Dot Slides for applications in microscopy and microanalysis.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Klaus Kemp at http://www.diatoms.co.uk/index.html makes stunning arrangments of diatoms and seems willing to share his knowledge.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu}
} } Hi: } } Does anyone know how to mount large (100um+) diatoms for LM or SEM? The } little ones are OK, they stick to most any stub, but these big guys like to } fall off. } } Anyone know how arranged diatom slides are/were made? I checked WWW but no } specifics on techniques. Something like these methods would work for us. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
Never cut myself in over quarter a century of trimming. Any accidental cuts would be minor since little force is exerted during final trimming. Some force is required during rough trimming and when cutting PE moulds off blocks - without a press. For these operations, just think where a slipped razor blade may go - and that is were the fingers don't. Surprizingly hardly anybody in my labs ever cut a finger, because they were taught the obvious. I think its also obvious, that unless those Kevlar gloves are rather thick (no analogy to the safety police) they would not prevent cuts. They should have recommended those stainless steel chain linked butcher's gloves. That could be a saleable idea! Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, January 11, 2001 10:03 AM, Calvert, Dave [SMTP:calvert-at-eastman.com] wrote: } } } } Listers } My company's safety folks are bent on making us wear Kevlar gloves while } razor trimming samples for ultramicrotomy - I'm talking about the final } trimming done using a trimming stand on the microtome. As you might know, } this is fine work - wearing gloves is a handicap. } My questions: } 1. Does anyone out there actually wear gloves? - if so then can you } recommend a glove? } 2. Are there other ways to make this task look less dangerous to the } safety police? (short of purchasing a trimming machine) } } Thanks! } } Dave Calvert } Eastman Chemical Company } Building 150B Lincoln Street (packages) } P.O. Box 1972 (US Mail) } Kingsport, Tennessee 37662-1972 } (423) 229-4943 } (423) 229-4558 fax } }
I have always worn latex gloves more for protection ( abrasion from tightening chucks, etc.) Using long single edge razors (Weck Extra Long Blades, EMS #71933-60) seems to give me more control of the blade and therefore less of a chance to knick one's fingers.
Becky Garrison Pathology Supervisor Shand Jacksonville Jacksonville, FL 32209 904-244-6237 -----Original Message----- } From: Calvert, Dave [mailto:calvert-at-eastman.com] Sent: Wednesday, January 10, 2001 7:03 PM To: Microscopy-at-sparc5.microscopy.com
Listers My company's safety folks are bent on making us wear Kevlar gloves while razor trimming samples for ultramicrotomy - I'm talking about the final trimming done using a trimming stand on the microtome. As you might know, this is fine work - wearing gloves is a handicap. My questions: 1. Does anyone out there actually wear gloves? - if so then can you recommend a glove? 2. Are there other ways to make this task look less dangerous to the safety police? (short of purchasing a trimming machine)
Thanks!
Dave Calvert Eastman Chemical Company Building 150B Lincoln Street (packages) P.O. Box 1972 (US Mail) Kingsport, Tennessee 37662-1972 (423) 229-4943 (423) 229-4558 fax
I have used LKB (now Reichert-Jung or whatever) microtomes and it has always been possible to trim specimen blocks with glass knives on the microtome, itself. With a bit of practice you can both trim and cut within about 30 minutes per block so I don't think that it's particularly time consuming. The added bonuses are that blocks will always have smooth facets and students get some good hands-on training with microtome, binoculars and knives before actually cutting thin sections.
Once you get into the technique there are other rewards such as the ability to 'face' or smooth block fronts for best quality sections on difficult specimens, mesa trimming, combining trimming of blocks and light microscopy surveys etc. The only requirements are that your specimen must be embedded near to the surface of the resin for easy trimming and that you don't damage the microtome mechanism.
The basic technique is fairly intuitive you just need practice: trim the front of the block until you reach the specimen (1-5 um cuts on manual feed should be OK) rotate the knife stage by +10 to +40 deg (angle is optional and can be varied for each side) and trim until you reach the desired edge in your block rotate knife stage to the other side (i.e. -10 to -40deg) and again trim until you have a long thin block face rotate block in specimen holder by 90deg and repeat for the other two edges Finally I may carefully trim or 'face' the front of the block for best results If you want to produce mesas then don't rotate the knife - just use it to trim a few microns around each side of the area to stand 'proud' as a mesa. If the the block is difficult and blunts the knife you may need to use a second knife, although this is not common Smoothest trimming results from thinnest cuts at slowest speeds, so always make the last couple of cuts on each edge about 1um or less.
If you're uncertain about the ability of your ultramicrotome to trim in this fashion, I'm sure the manufacturer or someone on the list should be able to help. If I do ever trim with a blade I use single edged razors and cut away from myself on an uncluttered work area, but this may only happen once or twice a year often when I'm demonstrating the method to students.
I'm sure that this has been mentioned on the list quite some time back but please contact me directly if you have any further questions although there must be lots of labs that have experience of this technique.
DISCLAIMER I have no commercial interest in LKB or Reichert, they just happen to be the instruments that I have grown up with.
Good luck
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My company's safety folks are bent on making us wear Kevlar gloves while } } razor trimming samples for ultramicrotomy - I'm talking about the final } } trimming done using a trimming stand on the microtome. As you might know, } } this is fine work - wearing gloves is a handicap. } } My questions: } } 1. Does anyone out there actually wear gloves? - if so then can you } } recommend a glove? } } 2. Are there other ways to make this task look less dangerous to the } } safety police? (short of purchasing a trimming machine) } } When I train people to trim blocks, I always point to the Band-Aids first, } then to the razor blades! This gets a laugh, but drives home the } point. Then I suggest that they put the bandages on FIRST before trimming, } which also gets a laugh, but significant number of people decide to do } so! Therefore, I suggest something like finger cots or other wrap-around } solutions that leave some dexterity while protecting the last joints of } the fingers. } } Aloha, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
I am trying to find a way of looking at a cross section of a printed ink approx 10um thick on a PVC substrate. I am trying to measure the porosity of the ink so I guess some image analysis will be used. It is mainly a method of preparing the samples that has me a bit stumped. I have tried freeze fracturing and embedding the samples in resin then polising the surface. Neither are satisfactory. The microscopy techniques I have available are SEM and LM. Can anyone help?
Largish diatoms, like most other microfossils bigger than, say, 50 microns or so in diameter or length, can usually be handled under a binocular microscope with a very fine (00 or 000) artists' paintbrush. Just wet the brush with water (distilled, if it's going to matter) and touch the bug with it. It'll stick to the brush long enough for you to place it on a stub. When I'm looking at microfossils in the SEM, I can usually get them to stick to double-sided tape on a stub just by "dabbing" them on with the point of the brush. Orienting them can be a bit tricky, but you'll improve with practice. One problem with double-sided tape, though, is that once stuck on, a lot of the more delicate microfossils such as diatoms will break before they'll come loose. So you don't want to handle type specimens this way.... Another good way to stick such bugs down includes a little bit of Gum Tragacanth. An old micropaleontologists' standby, this stuff is available as a powder, which you mix with a little water to make a paste. You can glue individual bugs down with this, using your fine paint brush. It dries in a few seconds - and if you want to move/remove the bug later, just wet your brush and touch the bug. The gum redissolves and the specimen comes free. Unsightly gum residues on the bug will come off with a bit more application of your wet brush. Once mixed up and kept in a sealed little vial, the gum mixture should keep for months or years. A drop or two of clove oil in it will inhibit any possible mould growth, if you're going to keep it around for a while. One guy I used to work with liked to use bits of old undeveloped 35mm film. Apparently you can get small items to stick to such film really well if the specimen is wet. I've never tried this myself, though. And, of course, there's lots of micropaleontology texts out there with chapters on handling techniques. After reading all this, you're probably sorry you got me started. Anyway, good luck with the diatoms - they're among nature's most photogenic micro-organisms.
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 10, 2001 4:54 PM
Hi Folks,
Does anyone know of a stain to differentiate PMN's from Macrophages for light microscopy?
for a 7 Hr Minicourse in Cryomicroscopy and Microanalysis
at the Microscopy Technology Center of San Joaquin Delta College in Stockton, CA
Everyone is welcome. Come, enjoy, and RSVP. Happy New Year
Delta Microscopy Society, & Northern California Microscopy Society Joint Meeting
WHEN: Jan 17, 18, 2001; Wed. and Thur.
WHERE: San Joaquin Delta College, Microscopy Technology Center, Stockton, CA
Please email or fax RSVP. See form below.
7 Hr Minicourse in Low Temperature Microscopy and Microanalysis by Dr. Patrick Echlin, Director of Cambridge Analytical Microscopy and member of Multi-Imaging Centre, University of Cambridge, UK
Image Database (Asset Management System) for Multi-Platform, Multi-Instrument Microscopy Laboratory, Dr Judy A. Murphy, San Joaquin Delta College, Microscopy Technology Center
January 17, 2001: Wednesday 12:00 - 1:00 pm Registration, Holt 121 1:15 - 1:30 pm Welcome and Introduction, South Forum Building 1:30 - 2:30 pm Minicourse Lecture 1 : Chemical-physics of water and ice 2:45 - 3:30 pm Minicourse Lecture 2: Sample cooling 3:30 - 4:30 pm Tour of Microscopy Technology Center, Holt 121 including Demo of Image Database 4:30 - 5:30 pm Minicourse Lecture 3; Sectioning and Fracturing Discussion Social 5:30 - 6:30 pm, Budd Lounge Light Buffet 6;30 - 8pm, Budd Lounge 8:00 - 9:00 Image Database Lecture
January 18, 2001: Thursday 8:00 - 8:45 am Registration, 121 Holt 9:00 - 10:00 am Minicourse Lecture 4: Freeze drying, freeze substitution and low temperature embedding 10:15 - 11 am Minicourse Lecture 5: Low temperature microscopy Discussion 11:15 - 1:00 Lunch 1:15 - 2:15 pm Minicourse Lecture 6: Low temperature microanalysis 2:30 - 3:30 pm Minicourse Lecture 7 : Imaging, artefacts and beam damage. Discussion
Any building or room changes will be posted in Holt 121.
Activities are being sponsored by: Gatan Inc., Oxford Instruments, Hitachi, JEOL, FEI, Ted Pella Inc., EMS/Diatome, Anatech, Cressington, LEO, Advanced Computers and Networking, Delicato Winery, Paul De Georgio and others to be identified later.
Please RSVP so we know what room sizes we will need, # of handouts, and how much food.
Cost to Students: $0 Cost to Other Registrants $15 (to defray cost of activities, handouts, etc.)
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Please email or fax RSVP
email to: murphyjudy-at-mediaone.net
FAX to: 209/954-5600 This is a central fax, so put Judy Murphy, Microscopy on top of fax.
Any questions, call Judy Murphy (209/954-5284) or Pat Kysar (209/954-5246)
Checks can be made out to Delta Microscopy Society and sent to: Judy Murphy, San Joaquin Delta College, Microscopy Technology Center, 5151 Pacific Ave, Stockton, CA 95207
Directions to Microscopy Technology Center & Lodging Holt Center San Joaquin Delta College 5151 Pacific Avenue Stockton, CA 95207
1. Find Interstate 5 (I-5) on California map. It is a main North/South Interstate in California. 2. From I-5, exit on March Lane going East. Continue on March Lane 1.6 miles until you reach the traffic light at Pacific Ave. 3. At Pacific Ave.turn LEFT, and go 0.3 miles until you reach the traffic light for Delta College. 4. Turn LEFT at Delta College Entrance. The street is labeled Yoguts going East and Delta College going West. 5. As soon you turn into Delta College Entrance, bear LEFT on the Campus Drive (called Burke-Bradley in some places). 6. Continue around on Campus Drive until you reach Holt 1 Parking Lot. Turn RIGHT into Holt 1 Parking Lot. 7. As soon as you turn into Holt 1 Parking Lot, take a sharp RIGHT turn to enter Holt 2 Parking Lot. 8. Park as close to the front of the parking lot as possible, closest to the buildings. 9. Since this is the first week of Spring semester classes, no tickets are issued, so you do NOT need a parking permit. 10. As you walk towards the buildings, you should first enter Holt building. Go to Holt 121 which is the Microscopy Center. 11. Any questions, call Judy Murphy (209/954-5284) or Pat Kysar (209/954-5246)
Lodging Within 1.5 miles of San Joaquin Delta College Prices were quoted Dec. 20, 2000, but check when you make reservations. All listed lodging is close to March Lane exit off Interstate 5
Red Roof Inn, 2654 March Lane, Stockton, CA, Phone: 209/478-4300 Fax: 209/478-1872; Rates: Single - $59.99; Queen or 2 Double - $65.99; King - $71.99; Suite - $87 and up depending on number of people.
LaQuinta, 2710 W. March Lane, Stockton, CA, Phone: 209/952-7800, Fax: 209/472-0732; Rates: King-$75; 2 Double Beds-$69, Complimentary Breakfast AAA, AARP,Senior Citizen Discount Rates: King-$71.25; 2 Double beds-$65.55
Radisson Hotel, 2323 Grand Canal Blvd., Stockton, CA, Phone 209/957-9090 Fax: 209/473-0739; Rates: $79 - $99. Canal Blvd is one block off March Lane.
Courtyard by Marriott, 3252 W. March Lane, Stockton, CA, Phone 209/472-9700, Fax: 209/472-9722; Rate: $89-104 . Marriott Residence Inn, 3240 W. March Lane, Stockton, CA, Phone: 209/472-9800 Fax: 209/472-9888; Rates: Studio - $119; 1 Bedroom - $124. Complimentary Breakfast every day. Rooms have Kitchens equipped with Stove, refrigerators, pots & pans etc
It sounds like the safety folks may be overzealous in their attempts to protect you from embedding resins. Once polymerized, the resins are considerably safer than the monomers. Since the plastic is no longer liquid, latex gloves would be more appropriate (if the individual is not allergic to latex).
A bigger concern, I would think, when working with polymerized plastic is the possible allergic reaction due to inhalation of the plastic chips. I strongly recommend the use of a dust mask if you are using a grinder or other mechanical device to shape the blocks.
} My company's safety folks are bent on making us wear Kevlar gloves while } razor trimming samples for ultramicrotomy - I'm talking about the final } trimming done using a trimming stand on the microtome. As you might know, } this is fine work - wearing gloves is a handicap. } My questions: } 1. Does anyone out there actually wear gloves? - if so then can you } recommend a glove? } 2. Are there other ways to make this task look less dangerous to the } safety police? (short of purchasing a trimming machine) -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
I have no experience in EDS detector pumping, but a little bit in vacuum technology.
What do you think about heating the Dewar before pumping ?
That's what I do with sorbtion (zeolite) pumps used to prepump UHV vessel (to have a dry pumping before starting turbo or ion pump). I put a heater around the pump -in our case it would be hot water (hot air ? hot oil ? which temperture can support the Dewar assembly ?)- let it warm over the night, and close the venting valve.
You could do something similar with the detector : warm it at air pressure for a few hours (it schould dry too the zeolite) and than pump down with turbo, but pre-pump with dry backing pump to prevent oil contamination, as Jim mentioned. Than pump down until the Dewar is cold, with a base pressure of something like 10E-5 Torr. And after that, cool it with nitrogen.
What do you think about that ?
Bye
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
} } } } My company's safety folks are bent on making us wear Kevlar gloves while } } razor trimming samples for ultramicrotomy } } 2. Are there other ways to make this task look less dangerous to the } } safety police? (short of purchasing a trimming machine)
We routinely use a Dremel moto-tool (available at your local hardware store) with thin cutting wheels (i prefer the ones from Small Parts, Inc that have an abrasive on one side and a very thin metal backing on the other side). Working in the hood to avoid the dust, we can trim dozens of blocks rapidly. I advise holding the block in a pliers in case you slip with the dremel but it really is very easy once you get used to it. -- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
If someone in your lab has a Perrot Fabry spectrometer, you can use it to mesure film thickness between 2 and 100 nm. You évaporate your film on a glass slide (LM slide), than you put a fine scratch on the film, than you evaporate a opticaly opaque film of silver or aluminum. Than on the Perrot Fabry, with a monochromatic light, you can see and mesure the hights of the siver setp, which covers the the film to bee mesured. You must know the wavelength of your light. You schould find the way to calculate this in any optics book. You must do a serie of different thickness to draw a calibration curve. Its very sensitiv and cheep,... if you have the Perrot Fabry of course
An other way, opticaly too, is on a LM with a Nomarski Prism. I used it 15 years ago, but I do know the way to make the optical montage. I remember that there is a monochromatique light, the Nomarski prism, the polariser and analyser, but what kind of objectif ? The sample must be scratched and silver covered as with the Perrot Fabry...
Can someone tell me something about that ?
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I had a quick look at the specification of the Coolscan 8000ED and it seems to be aimed at TEM users with about 6x9cm films - I quote the largest areas from the web page: (6 x 9) 63.5 x 88.0mm (10,000x13,176 pixels); (Elect Micro) 56.9 x 83.7mm (8,964x13,176 pixels); My e.m. film is 4 x 3 5/16 inches or 83 x 102 mm with an absolute minimum picture area (without data) of 69 x 92mm so it may be a problem for some e.m.
users.
I must admit it's a great specification though.
Malcolm Haswell e.m. unit University of Sunderland UK
George Laing wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } All, } The preliminary information I have is that the Coolscan 8000 ED } will sell for approximately $4000 US. We will carry it when it becomes } available, currently listed as June. I hope to see it in early February. } Quick specs.. } 4000dpi optical resolution } IEEE1394 "Firewire" Interface- not SCSI } 4.2 dynamic range 48bit images } Multi Sample Scanning } } Info is also at } http://64.77.39.20/usa_product/product.jsp?cat=7&grp=703&productNr=9246 } } I'll pass along more info as it becomes available. } } George } } George Laing } National Graphic Supply } v:(800) 223-7130 X3109 } f:(800) 832-2205 } email: scisales-at-ngscorp.com } } } } This looked pretty interesting. Have you heard a ballpark price? } } } } } } } } In the context of recent queries, I just thought I'd } } } post Nikon's most recent announcement ... see: } } } http://www.klt.co.jp/Nikon/Press_Release/ls-8000.html } } } } } }
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Reply to: RE: Ultramicrotomy safety issues It is possible to perform all the trimming steps needed to get a good block by using glass knives. Not only is it safer for the operator it also produces blocks with smooth, regular edges which are great for serial sectioning. More details provided on command.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Calvert, Dave wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listers } My company's safety folks are bent on making us wear Kevlar gloves while } razor trimming samples for ultramicrotomy - I'm talking about the final } trimming done using a trimming stand on the microtome. As you might know, } this is fine work - wearing gloves is a handicap. } My questions: } 1. Does anyone out there actually wear gloves? - if so then can you } recommend a glove? } 2. Are there other ways to make this task look less dangerous to the } safety police? (short of purchasing a trimming machine) } } Thanks! } } Dave Calvert } Eastman Chemical Company } Building 150B Lincoln Street (packages) } P.O. Box 1972 (US Mail) } Kingsport, Tennessee 37662-1972 } (423) 229-4943 } (423) 229-4558 fax } } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A8F3168B0262; Wed, 10 Jan 2001 22:55:47 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id SAA00552 } for dist-Microscopy; Wed, 10 Jan 2001 18:06:37 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id SAA00549 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 10 Jan 2001 } 18:06:07 -0600 (CST) } Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id SAA00542 } for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 10 Jan 2001 18:05:55 -0600 (CST) } X-Sender: zaluzec-at-ultra5.microscopy.com } Message-Id: {v03130301b682a8bd6c36-at-[206.69.208.21]} } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Wed, 10 Jan 2001 18:03:27 -0600 } To: Microscopy-at-sparc5.microscopy.com } From: "Calvert, Dave" {calvert-at-eastman.com} } Subject: Ultramicrotomy safety issues } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273278871 } Status: R }
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA02680 for dist-Microscopy; Thu, 11 Jan 2001 11:45:47 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA02677 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 11 Jan 2001 11:45:17 -0600 (CST) Received: from utilitysrvr.qdots.com ([207.33.253.219]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id LAA02670 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 11 Jan 2001 11:45:06 -0600 (CST) Received: by UTILITYSRVR with Internet Mail Service (5.5.2448.0) id {CN74FMKY} ; Thu, 11 Jan 2001 09:25:52 -0800 Message-ID: {4B747C1B1138D211B9DB0008C7BF0B242E026F-at-UTILITYSRVR}
Dear Colleagues,
Does anyone know any place that has a TEM with a dedicated STEM unit attached. The reason for the question is that I have particles composed of cores and shells (coated particles), both of which are different materials. By TEM I am not able to discern the shells from the cores because the contrast is extremely low. However, I am told that a dedicated STEM unit might allow me distinguish the shells from the cores based on the fact that they both have very different z-contrast.
Regards, Thearith
_________________________
Thearith Ung, Ph.D. Quantum Dot Corporation 26136 Research Road Hayward CA 94545 Tel: (510)-887-8775 (x4125) Fax:(510)-783-9729 www.qdots.com
Hi, I'm looking for technical assistance or a service manual for a Leitz Orthomat E (7916) Photo System.The problem we are having is the unit will not allow a picture to be taken unless there is very little light. Once the system thinks it is in range, there is not enough light present to get the photograph. An adjustment of the light detector? Any assistance is appreciated. Lewis McCrigler Humboldt State University
I've never worked with ink samples, but nevertheless I am trying to give some advice (be cautious!).
I think that techniques like freeze fracturing are not suitable for your purposes. Cracks could prefer to propagate through pores, so fracture surface will exaggerate porosity. Polished cross sections are much better for quantification. Maybe you could find suitable media for embedding. And what about mounting 2-3 substrates in a clip and cutting them with a sharp blade (hm... I really do not know ink's properties...)?
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Tiffany Argent [mailto:tiffany.argent-at-abbott.com] } Sent: Thursday, January 11, 2001 6:07 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Cross section sample preparation } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } I am trying to find a way of looking at a cross section of a } printed ink } approx 10um thick on a PVC substrate. I am trying to measure } the porosity of } the ink so I guess some image analysis will be used. It is } mainly a method } of preparing the samples that has me a bit stumped. I have } tried freeze } fracturing and embedding the samples in resin then polising } the surface. } Neither are satisfactory. The microscopy techniques I have } available are } SEM and LM. Can anyone help? }
One of our investigators is studying the ratio of total and phosphorylated transcription factor in the cell. We are able to do multiple fluorescence labeling but I need help in determining the fluorescence intensity ratios. We are using the Alexa dyes 350, 488 and 630 just to make sure there is no bleed through with the filters that we use. I would like to know if anyone has done this type of experiment and what are the potential problems that one could run into. I would appreciate comments or suggestions.
Thank you,
Cora Bucana Corazon D. Bucana, Ph.D. Dept. Cancer Biology UT M.D.Anderson Cancer Center 1515 Holcombe Blvd, Box 173 Houston, Texas 77030 Phone: 713-792-8106 FAX 713-792-8747
We've only lost about 4 technologists due to them accidentally cutting open their jugulars with a razor blade whilst trimming blocks and bleeding to death on the spot. So with that good safety record, we've chosen to dispense with the gloves thus far. Should we lose a few more though, the issue might come up again.
Garry
Charge Technologist - Electron Microscopy Department of Pathology Health Sciences Centre Winnipeg, Canada
I've never done any serious particle arranging, but perhaps one of the people mentioned below can help you.
You might try asking Klaus Kemp via the website at http://www.diatoms.co.uk since he creates arranged slides as a profession. He has been referred to as a master of the art at several microscopy seminars I've attended.
http://www.microscopy-uk.org.uk/mag/artapr00/machslide.html will take you to an article titled "A microscopical view of an old diatom circle pattern slide with 250+ diatom shells" by Martin Mach (there is a link to his e-mail). He provides a "Modern Microscopy" reference for preparation of such slides, but unless you have a GOOD library, you might ask him if he could send you a copy of the article - it is dated 1895.
Anna Teetsov at McCrone Associates (http://www.mccrone.com/ma/staff.html) is also expert at positioning particles on slides. While she is famous for manipulating the tiniest of particles, I've seen some beautiful arranged butterfly scale slides she's made for viewing under the light microscope. She might be willing to provide some suggestions.
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-339-7300 phone 508-339-4996 fax Louise_Harner-at-albint.com
jmkrupp-at-cats. ucsc.edu (Jon To: Microscopy-at-sparc5.microscopy.com Krupp) cc: Subject: Mounting large diatoms? 2001/01/10 03:54 PM
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Hi:
Does anyone know how to mount large (100um+) diatoms for LM or SEM? The little ones are OK, they stick to most any stub, but these big guys like to fall off.
Anyone know how arranged diatom slides are/were made? I checked WWW but no specifics on techniques. Something like these methods would work for us.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
A colleague (visiting from Brazil) is looking for a microtome for cutting semi-thin (0.25 to 2 micrometer) sections of plastic (epoxy) embedded specimens (mammalian tissues). He is considering a new Leica RM2165 ($18K).
I am soliciting testimonials (good and bad) for this or ANY other microtomes for cutting semi-thins.
Thank you on his behalf.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
We've solved that problem with Kevlar collars. We found that gloves just made it MORE likely that jugulars would be severed, so we don't use them either.
Randy
-----Original Message----- } From: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, January 11, 2001 1:19 PM To: microscopy-at-sparc5.microscopy.com
We've only lost about 4 technologists due to them accidentally cutting open their jugulars with a razor blade whilst trimming blocks and bleeding to death on the spot. So with that good safety record, we've chosen to dispense with the gloves thus far. Should we lose a few more though, the issue might come up again.
Garry
Charge Technologist - Electron Microscopy Department of Pathology Health Sciences Centre Winnipeg, Canada
In my 21 years of thin sectioning I have from time to time cut my fingers, but a band aid is all that was needed. Kevlar gloves sound to me like ridiculous over kill. However for those of us working in really big and not always friendly cities, there might be an argument for Kevlar vests. Happy sectioning, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
----- Original Message ----- } From: "Garrison, Becky" {becky.garrison-at-jax.ufl.edu} } I have always worn latex gloves more for protection ( abrasion from } tightening chucks, etc.) Using long single edge razors (Weck Extra Long } Blades, EMS #71933-60) seems to give me more control of the blade and } therefore less of a chance to knick one's fingers. } I have at times shaved with a straight razor and that is little different that what a long single edge razor. Some people that have beards still do although a old Gillette safety razor with the guard removed is easier to use since sharpening a straight razor is a pain. Explaining that some folks still use them to shave should at least catch the safely Nazi off guard if not convince him of the absurdity of his request.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Hi! I am interested in using a light microscope to digitize photographic film, i.e. to use my light microscope as a film scanner to scan negatives.
Since our lab does a fair amount of investigation into telepathology applications, we have scopes that are set up for the automatic scanning of multiple fields, merging of fields, etc. That problem is (relatively) solved.
What I am getting stumped on here is how to mount my 35mm photographic negatives. If I just slap one on a slide and set a coverslip on top, I get (not surprisingly) nontrivial refractive problems with irregularities in the surface of the film.
Unfortunately, these are photographic negatives which might have forensic/evidentiary value, so I can't dribble on any of the mounting media I can think of -- since I can't permanently mount the negative. I have to be able to take that negative at a later time and make an "original" photograph.
Anybody have experience with this? Any lore would be appreciated.
A one day TEM course sponsored by the AVS will given during the upcoming February Santa Clara meeting. For more information please see www.vacuum.org
At 9:04 AM -0800 12/13/00, Thearith H. Ung wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Glycerin has about the right refractive index and is easily washed out with water, after which the negatives can be air dried. Certainly glycerin works as a temporary mountant for microscope slides and the negative's gelatin emulsion is not affected by it. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, January 12, 2001 5:30 AM, William R. Oliver [SMTP:oliver-at-cpt.afip.org] wrote: } } } } Hi! I am interested in using a light } microscope to digitize photographic film, i.e. } to use my light microscope as a film scanner } to scan negatives. } } Since our lab does a fair amount of investigation } into telepathology applications, we have scopes } that are set up for the automatic scanning of } multiple fields, merging of fields, etc. That } problem is (relatively) solved. } } What I am getting stumped on here is how to } mount my 35mm photographic negatives. If I } just slap one on a slide and set a coverslip } on top, I get (not surprisingly) nontrivial } refractive problems with irregularities in the } surface of the film. } } Unfortunately, these are photographic negatives } which might have forensic/evidentiary value, so } I can't dribble on any of the mounting media I } can think of -- since I can't permanently mount } the negative. I have to be able to take that } negative at a later time and make an "original" } photograph. } } Anybody have experience with this? Any lore } would be appreciated. } } } billo } }
Don't do it. The reason for pumping detectors first without heat is to avoid further contamination of the crystal. When the dewar is heated material is released from the walls of the contained space and especially the molecular sieve. Pump first and then heat, that way the pump can cope and quickly eliminate the additional contaminants. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, January 12, 2001 2:28 AM, Faerber Jacques [SMTP:Jacques.Faerber-at-ipcms.u-strasbg.fr] wrote: } } Hello } } I have no experience in EDS detector pumping, but a little bit in vacuum } technology. } } What do you think about heating the Dewar before pumping ? } } That's what I do with sorbtion (zeolite) pumps used to prepump UHV vessel } (to have a dry pumping before starting turbo or ion pump). I put a heater } around the pump -in our case it would be hot water (hot air ? hot oil ? } which temperture can support the Dewar assembly ?)- let it warm over the } night, and close the venting valve. } } You could do something similar with the detector : warm it at air } pressure for a few hours (it schould dry too the zeolite) and than pump } down with turbo, but pre-pump with dry backing pump to prevent oil } contamination, } as Jim mentioned. Than pump down until the Dewar is cold, with a base } pressure of something like 10E-5 Torr. And after that, cool it with } nitrogen. } } What do you think about that ? } } Bye } } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Materiaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess } 67037 Strasbourg CEDEX } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
There is also a film cleaning fluid I found several at Porters Camera http://www.porters.com/ and search for "film cleaner" in their on line store. These usually don't require washing and would be easier to clean off than Glycerin. There is a scratch remover that might also work. I couldn't find it so it may have had something in it that the EPA didn't like. Your darkroom or local camera store may have these.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 } From: "Jim at ProSciTech" {jim-at-proscitech.com} } } Glycerin has about the right refractive index and is easily washed out with } water, after which the negatives can be air dried. Certainly glycerin works as } a temporary mountant for microscope slides and the negative's gelatin emulsion } is not affected by it. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Friday, January 12, 2001 5:30 AM, William R. Oliver } [SMTP:oliver-at-cpt.afip.org] wrote: } } } } } } } } Hi! I am interested in using a light } } microscope to digitize photographic film, i.e. } } to use my light microscope as a film scanner } } to scan negatives. } } } } Since our lab does a fair amount of investigation } } into telepathology applications, we have scopes } } that are set up for the automatic scanning of } } multiple fields, merging of fields, etc. That } } problem is (relatively) solved. } } } } What I am getting stumped on here is how to } } mount my 35mm photographic negatives. If I } } just slap one on a slide and set a coverslip } } on top, I get (not surprisingly) nontrivial } } refractive problems with irregularities in the } } surface of the film. } } } } Unfortunately, these are photographic negatives } } which might have forensic/evidentiary value, so } } I can't dribble on any of the mounting media I } } can think of -- since I can't permanently mount } } the negative. I have to be able to take that } } negative at a later time and make an "original" } } photograph. } } } } Anybody have experience with this? Any lore } } would be appreciated. } } } } } } billo } } } } } }
does anybody knows the telephone number from Biosupplies (Melbourne, Australia), I bought a callose ((1-3)beta-glucan-specific) monoclonal antibody two years ago and I can't contact with them at the telephone that I have. Thanks a lot Nuria
------------------------------------------------------------------------------ Nuria Cortadellas Unitat de Microscopia de Transmissio Serveis Cientifico-Tecnics Universitat de Barcelona C/ Sole i Sabaris, 1-3 08028 Barcelona Tel: +34 3 402 13 52 Fax: +34 3 402 13 98 E-mail: nuriac-at-giga.sct.ub.es
We put up with the minor wounds. I do however tell students not to cut themselves as I will be obliged to fill in an accident form and the safety officer will invite me to think of alterative methods. We thought of blunting the blade corners or making little corner guards but never got round to it.
Dave
On Thu, 11 Jan 2001 15:50:36 -0500 Timothy Schneider {Timothy.Schneider-at-Mail.TJU.EDU} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dave: } } In my 21 years of thin sectioning I have from time to time cut my fingers, } but a band aid is all that was needed. Kevlar gloves sound to me like } ridiculous over kill. However for those of us working in really big and not } always friendly cities, there might be an argument for Kevlar vests. Happy } sectioning, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
we use a Hitachi H7000 with STEM attachment although I'm sure there will be someone much closer to you, but I would like to make a couple of comments: you mention a dedicated STEM unit which to me is a free-standing high resolution STEM and not a STEM attachment to a TEM. There should be lots of TEMs with STEM attachments on instruments bought in the last 10 or 15 years but only a few high resolution STEMs because they are expensive and specialized (try Brookhaven National Laboratory website http://bnlstb.bio.bnl.gov/biodocs/stem/stem.htmlx).
You may also want to consider that certain TEMs have built in energy filtering or electron energy spectrometers which can actually be tuned for different atomic masses in the specimen - Zeiss EM902s can do this but I'm not sure if any other machines are available.
I have no connection with Brookhaven or Zeiss, although of course I do use Hitachi.
Good luck
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
"Thearith H. Ung" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Colleagues, } } Does anyone know any place that has a TEM with a dedicated STEM unit } attached. The reason for the question is that I have particles composed of } cores and shells (coated particles), both of which are different materials. } By TEM I am not able to discern the shells from the cores because the } contrast is extremely low. However, I am told that a dedicated STEM unit } might allow me distinguish the shells from the cores based on the fact that } they both have very different z-contrast. } } Regards, } Thearith } } _________________________ } } Thearith Ung, Ph.D. } Quantum Dot Corporation } 26136 Research Road } Hayward CA 94545 } Tel: (510)-887-8775 (x4125) } Fax:(510)-783-9729 } www.qdots.com
Glycerin, and also ethylene glycol, can be dried under vacuum making it possible to avoid rehydrating the negative Chris
----- Original Message ----- } From: "Jim at ProSciTech" {jim-at-proscitech.com} To: "'William R. Oliver'" {oliver-at-cpt.afip.org} ; {microscopy-at-sparc5.microscopy.com} Sent: Friday, January 12, 2001 12:52 AM
Before this subject is banned from the list -
Our only serious injuries to date are blinded students falling asleep while sectioning and poking their eyes out on the binocular eyepieces!
Perhaps the kevlar collars would keep their heads up a little longer?!
Pete
-- Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
I am trying to judge the optical quality of two similar stereoscopic zoom scopes: the Nikon SMZ800 and the Leica MZ7, both equipped with a 1x Planapochromatic objective.
Leica publishes resolution figures for the MZ7, claiming 309 line pairs per mm. Nikon does not, and I haven't been able to persuade the Nikon representative to "characterize" the resolution of the SMZ800, although he says he can. I do not have the necessary test slide, nor do I know where I can get one with a scale graduated from say 200-400 lp/mm. I do know though that such a slide would cost probably $300-$500.
Interestingly, Nikon publishes a resolution number for its next higher quality scope, the SMZ1000. It claims the SMZ1000 can resolve 300 lp/mm with a 1x Planapo. Because Nikon product literature doesn't publish a resolution for the SMZ800, I suspect that its resolution is substantially lower than 300 lp/mm, and therefore inferior to the Leica MZ7.
Does anyone have any suggestions on how I might judge the relative quality of these two scopes, or does anyone have experience with either scope that might shed light on this problem? Is my only alternative to find and buy a suitable test slide? If so, can anyone point me to a source?
Thanks very much, Ed
Ed Bachmann Odum Institute for Research in Social Science Manning Hall CB 3355 University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3355 (919) 962-0512
Here are a couple of references you might find useful, from a specialist on this kind of electron microscopy. I'll send the abstracts as attachments separately.
Srnova-Sloufova I, Lednicky F, Gemperle A, et al. Core-shell (Ag)Au bimetallic nanoparticles: Analysis of transmission electron microscopy images LANGMUIR 16: (25) 9928-9935 DEC 12 2000
Lednicky F, Coufalova E, Hromadkova J, et al. Low-voltage TEM imaging of polymer blends POLYMER
} "Thearith H. Ung" wrote: } } } } Dear Colleagues, } } } } Does anyone know any place that has a TEM with a dedicated STEM unit } } attached. The reason for the question is that I have particles composed of } } cores and shells (coated particles), both of which are different materials. } } By TEM I am not able to discern the shells from the cores because the } } contrast is extremely low. However, I am told that a dedicated STEM unit } } might allow me distinguish the shells from the cores based on the fact that } } they both have very different z-contrast. } } } } Regards, } } Thearith } } } } _________________________ } } } } Thearith Ung, Ph.D. } } Quantum Dot Corporation } } 26136 Research Road } } Hayward CA 94545 } } Tel: (510)-887-8775 (x4125) } } Fax:(510)-783-9729 } } www.qdots.com
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Abstract: Layered core-shell bimetallic silver-gold colloids in the size range of 10-16 nm have been prepared by the seed-growth method. Silver nuclei were covered by gold shells of various thicknesses without any stabilization agent. Interfacial (Ag)Au colloid-2,2'-bipyridine films were prepared from these bimetallic colloids and used for the purpose of analysis of transmission electron microscopy (TEM) images and electron diffraction. Both observed and calculated TEM images were used to characterize the prepared nanoparticles. On the basis of the analysis of TEM images, the calculated TEM image contrast, and results obtained by electron diffraction, energy-dispersive X-ray analysis, and other experiments, the core-shell structure of the prepared (Ag)Au nanoparticles was revealed. Particles were found to consist of a silver core and a gold shell enriched with silver.
Abstract: Low-voltage transmission electron microscopy (LV-TEM) was applied to obtain images of the phase structure of selected polymer blends without any prior staining. The instrument used (LVTEM-5, working at 5 kV) is of a novel construction combining visual-light and electronmicroscopical techniques, resulting in an enhanced efficiency of light transport to the eye and facilitating CCD imaging. Results were compared wi th LV-STEM at 25 kV. Phase structure of polycarbonate/poly( styrene-co-acrylonitrile) (PC/SAN), polystyrene/polypropylene (PS/PP), and polyethylene/polypropylene blends (PE/PP, ADFLEX) were selected to demonstrate the above techniques. The difference in density between the individual components of polymer blends was found to be the reason for the obtained image contrast. Differences less than 0.04 g/cm(3) can be traced with this technique.
First off, I can't understand the rationale for using an LM to do neg scanning. Why not just use a scanner? Easy, fast, reproducable.
If you do want to use an LM, it seems to me that a low power objective is required in order to avoid imaging mostly the grain structure. With a typical field of view for the camera, you will likely be taking about 100 individual shots and then stitching them together. A 2.5X objective works on low NA condensers. A 1X objective requires an ultra low condenser. But even this would result in a 10X FOV on each image frame.
Notwithstanding the above, if you do want to temporarily mount 35mm frames, get Bair mounts from the Stock Solution, Salt Lake City.
http://www.tssphoto.com/sp/index.html
These are self-adhesive cardboard mounts which stick to themselves but not to the film. When you are done, just peel the mount apart and your neg will fall out.
gary g.
http://photoweb.net
At 11:29 AM 1/11/01, you wrote:
} Hi! I am interested in using a light } microscope to digitize photographic film, i.e. } to use my light microscope as a film scanner } to scan negatives. } } Since our lab does a fair amount of investigation } into telepathology applications, we have scopes } that are set up for the automatic scanning of } multiple fields, merging of fields, etc. That } problem is (relatively) solved. } } What I am getting stumped on here is how to } mount my 35mm photographic negatives. If I } just slap one on a slide and set a coverslip } on top, I get (not surprisingly) nontrivial } refractive problems with irregularities in the } surface of the film. } } Unfortunately, these are photographic negatives } which might have forensic/evidentiary value, so } I can't dribble on any of the mounting media I } can think of -- since I can't permanently mount } the negative. I have to be able to take that } negative at a later time and make an "original" } photograph. } } Anybody have experience with this? Any lore } would be appreciated. } } } billo
Some things don't need solvents. For example, citric acid can be melted on a hotplate at about 100^C, and if cooled slowly will give chunky crystals in all sorts of birefringence colours.
If you have a hot enough plate, it's good to look at sodium nitrate (mp about 305^C. This will form large slabs of crystals. These don't look so good in themselves, but if you have a Bertrand lens or else simply pull out the eyepiece and look at down the tube, many of them will display concentric rings on a Maltese cross, the interference pattern resulting from different path lengths of the two polarized rays through the crystal. Try this with different magnification objectives. The citric acid will also give wonderful interference patterns, but they're much more complicated.
Other organic compounds can simply be melted, and when cooled between cover slip and hot plate will form spherulites.
If you melt a thin film of polypropylene between slide and cover slip, and cool slowly, you will get fantastic spherulites. Spherulites are found in many plastics, but polypropylene is the star performer. (Further details on request).
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
On Fri, 5 Jan 2001 us004118-at-mindspring.com wrote:
} Question: I am enjoying doing photomicrographs of crystal preparations } in polarized light. However I have insuffient knowledge of chemistry to } choose solvents without a long series of trial and error efforts.Can you } give me a rationale and/or a reference to go about this in a more } productive manner?
Email: us004118-at-mindspring.com Name: Leonard Lessin, FBPA
*************************************************** Last call for papers: FOCUS ON MICROSCOPY 2001 ***************************************************
FOM 2001 , Amsterdam The Netherlands, April 1-4, 2001
FOM 2001 is a world leading international conference on advanced microscopy. It is the joint meeting of the 13th International Conference on Confocal Microscopy and the 13th International Conference on 3D Image Processing in Microscopy.
FOM 2001 will be held at the Academic Medical Centre (AMC) of the University of Amsterdam, Amsterdam, The Netherlands.
For more information visit: http://www.focusonmicroscopy.org
Call for abstracts ------------------ Deadline for abstract submission: february 1st, 2001.
Core conference subjects: "INSTRUMENTATION" and "IMAGE PROCESSING AND ANALYSIS"
SPEAKERS: S. Hell (Gottingen, Germany), T. Wilson (Oxford, UK), S. Kawata (Osaka, Japan) C.J.R. Sheppard (Sydney, Astralia), C. Coggswell (Sydney, Astralia) E. Stelzer (Heidelberg, Germany) C. Cremer (Heidelberg, Germany), P.A. Benedetti (Pisa, Italy), A. Kriete (Giessen, Germany), H. van der Voort (Hilversum, The Netherlands), P.C. Cheng (Buffalo, USA), G.J. Brakenhoff (Amsterdam, The Netherlands)
Special conference subjects this year: MULTI-PHOTON MICROSCOPY" and "MICROSCOPY OF LIVING CELLS AND TISSUE".
SPEAKERS: A. Zumbusch (Muenchen, Germany), L. Moreaux (Paris, France) A. Diaspro (Genova, Italy), M. Muller (Amsterdam, The Netherlands), K. Sullivan (La Jolla, USA, to be confirmed), R. van Driel (Amsterdam, The Netherlands), R. Eils (Heidelberg, Germany), T. Mistelli (Bethesda, USA; to be confirmed), D. Gadella (Wageningen, The Netherlands), J. Dobrucki (Krakov, Poland), A. Houtsmuller (Rotterdam, The Netherlands), E. Manders (Amsterdam, The Netherlands)
} First off, I can't understand the rationale for using an } LM to do neg scanning. Why not just use a scanner? } Easy, fast, reproducable.
There are a couple. The driving reason for me is that this gives me a *lot* more control in exploiting the lighting of the negative.
I do forensic image processing as well as "regular" pathology stuff. Consider the following scenario:
A bad guy is sitting in a shadowed room aiming a rifle through a window. Joe Snuffy tourist takes a photo of the street which incidentally captures the window (let's assume that Joe Snuffy is a purist and isn't using a Mavica, in which this is all moot and useless). The window is largely a dark blob.
The task, then is to get as much information of that dark blobby window as possible. If you look at most film scanners on the market, particularly in any reasonable price range, you don't have the option of cranking your illumination up or down, of sticking filters in the light pathway, of doing all the stuff we take for granted as microscopists.
There's a lot of tools in them thar microscopy hills. It ain't so, it seems, with the film scanners I have available.
Instead, almost always, the only thing you can play with is the gamma. That means you are going to spend a lot of your representation space on stuff you don't want to see. The scanner is going to get two bits of data out of that blob, whatever knob you twiddle.
Playing games resetting the gamma as an option in a TWAIN driver won't fix this, and I haven't seen a "burn the bejezus out of this little area here and ignore the rest of the slide" button. Oh, sure, you can take those two bits the scanner gives you and display it as 0 and 1 or 0 and 255, but it ain't going to fill up the space in between. Not even with noise. The bottom line is that my impression is that film has dynamic range that is not accessible to most film scanners.
As a secondary issue, I have a few ideas for for playing with grain and restoration that might benefit from data aquisition at a higher resolution.
} } If you do want to use an LM, it seems to me that a low } power objective is required in order to avoid imaging } mostly the grain structure.
Sorta, but I am actually interested in seeing about characterizing the grain structure and seeing if that characterization can be useful in restoration.
Finally, I have a good characterization of the mtf of my microscope. I don't have a good characterization of the mtf of my flatbed scanner. While that doesn't help me with blind deconvolution for the camera mtf, it does help me for at least that part of the restoration of the image. I may be goofy, but I'd like to play with it a little.
If you have some experience with this, I'd love to learn from you. I figure that *somebody* must have played with this, but I haven't been able to find much literature on it. Do you have some lore you would like to share (nudge, nudge)?
} With a typical field of view } for the camera, you will likely be taking about 100 } individual shots and then stitching them together.
Something like that. But that stitching and data handling is something we do on a daily basis.
} } Notwithstanding the above, if you do want to temporarily } mount 35mm frames, get Bair mounts from the Stock } Solution, Salt Lake City. } } http://www.tssphoto.com/sp/index.html
} Does anyone know any place that has a TEM with a dedicated STEM unit } attached. The reason for the question is that I have particles composed } of cores and shells (coated particles), both of which are different } materials. } By TEM I am not able to discern the shells from the cores because the } contrast is extremely low. However, I am told that a dedicated STEM unit } might allow me distinguish the shells from the cores based on the fact } that they both have very different z-contrast.
I agree with the person who suggested an energy-filtering TEM, such as the Zeiss 902. We have the newer version, the LEO 912, and I have also been doing some work with coated nanoparticles. Yes, it is possible to see the difference between the cores and coating with this type of instrument.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Someone here would like to probe for heparin on some amyloid fibrils they have made. Fibril formation is different in the presence of heparin and they are curious to know if the heparin is binding to the fibrils or not.
They prepare the fibrils in their lab, don't ask me how, and apply liquid suspensions to a formvar coated grid. We negative stain with UA and then go to the TEM to look for fibrils.
The results of the neg stain search can be variable, sometimes the fibrils look like fibers, other times it seems to be just a bunch of junk, so the fibril formation is not always we defined. These folks are trying to figure out whether to probe the fibrils on the grid, or before while still suspended. But they tell me that often the fibrils do not survive too many washings in suspension, so maybe probing in solution before applying to grids could be a problem.
Any ideas, this is not my area of expertise.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Food Structure & Functionality Symposium 2001 An international symposium leading food structure & Functionality studies through the 21st century *webaddress at the AOCS site http://www.aocs.org/member/division/fsff/index.htm*
Being held in conjunction with the , May 13-16, 2001, Minneapolis Convention Center, Minneapolis, Minnesota, USA. For information, e-mail us at: meetings-at-aocs.org
The symposium has two themes: * New and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * Food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Program (confirmed as of January 12th, 2001)
May 13th - Short Courses (short courses will run for a full day, and will run concurrently)
1) Food Contaminants. Contact person - Mark Auty 2) Specific Labeling in Foods. Contact person - Marcel Paques
May 14th-16th inclusive - Technical sessions 6 sessions will run over 3 days
Morning Symposium opening Plenary lecture - Food Quality andwhy the Structure matters. P. Lillford, Unilever Research, Colworth House UK
Session 1: Dairy Products and Fat Based Foods Session - chairs M. Auty and M. Paques
Milk protein polysaccharide interactions in aqueous solutions and oil-in-water emulsions. H. Singh*, Y. Hemar & P. Munro, Institute of Food, Nutrition and Human Health Massey University, Palmerston North, New Zealand
Structural functions of dairy ingredients in products formulated with taro flour. C. Onwulata, USDA/ARS,Eastern Regional Research Center,Wyndmoor, PA 19038
Confocal imaging of galactomannan mode of action in recrystallisation of ice in model ice cream D. Ferdinando, Unilever Research Colworth House, UK
Heating of Food Proteins in a Closed System at High Temperature N. Kitabatake, Kyoto University, Japan
Milk Protein - molecular components and functional properties. N. C. Ganguli, Indian Dairy Association
Afternoon Session 2: Food Safety - chairs J. W. Arnold and R. Droleskey Prevention of Foodborne Illness Through Sanitation and Processing Technology J.W. Arnold; USDA/ARS, Russell Research Center; Athens, GA 30604
PreHarvest Intervention Strategies to Control Foodborne Pathogens in Poultry J. A. Byrd; USDA/ARS/SPARC; College Station, TX 77845
Interactions of Competitive Exclusion Cultures with the Intestinal Mucosa of Newly Hatched Chicks R. Droleskey; USDA/ARS/SPARC; College Station, TX 77845
Adhesion of Salmonella on Alfalfa Sprouts A. Chartowski; USDA/ARS, Western Regional Research Center; Albany, CA 94710
Growth of Fusarium moniliforme Dependent upon Corn Tissue Type I. E. Yates; USDA/ARS, Russell Research Center; Athens, GA 30604
Dedicated poster viewing 4:00-6:00PM
Evening: Round Table Discussion - topic to be announced
Tuesday, May 15th, 2001 Morning Session 3: New Methods and Techniques for Food Structure and Functionality Analysis -chair K.Groves
Diffusing wave spectroscopy - a new and non-invasive method for the investigation of the structure, dynamics and interactions in complex food systems M. Alexander* and P. Schurtenberger
Food: How complex can it be? E. Esselink ,Unilever Research Vlaardingen, The Netherlands
Immunolocalization of Transgenic Protein in Wheat Endosperm M. L. Parker *, E. Stoger, R. Casey and P. Christou, Institute of Food Research, Norwich, England.
Structure/Function relationships through microrheology. M. Paques, Wageningen Centre for Food Sciences/Unilever Research Vlaardingen
Specific labeling techniques for foods J. Leunissen, Aurion: Immuno-Gold Reagents & Accessories, The Netherlands
Food Structure and Functionality Division Luncheon. Dr. Brian Brooker (Institute of Food Research, England * retired) will be presented with the Food Structure and Functionality Division award and will give a presentation entitled: Fat crystals - the importance of being small.
Afternoon Session 4: Agricultural Applications of Microscopy and Imaging Session- chairs D.F.Wood and P. Allan-Wojtas
The Utility of Sorting in Agriculture. H. J. Arnott, Department of Biology, University of Texas at Arlington, Texas
The Potential for Automatic X-ray Sorting of Insect Infested Grain R. Haff . USDA - ARS - WRRC, Albany, California.
Automated Sorting of Almonds with Embedded Shell by Laser Transmittance Imaging T. Pearson* R. Young, USDA - ARS - WRRC, Albany, California.
Use of a GFP-transformed strain of Fusarium graminearum to study ear rot development in corn S. S. Miller. , AAFC - ECORC, Ottawa, Ontario, Canada.
Popping modifies endosperm structure and improves digestibility in maize and sorghum. M.L. Parker, Institute of Food Research, Norwich, England.
Microstructural Changes in Rice During Cooking D. Wood* and P.C. Yu . USDA - ARS - WRRC, Albany, California.
Evening: Food Structure and Functionality Division Member meeting
Wednesday, May 16th, 2001 Morning Session 5: Ingredients and Food Processing - chairs D. Kittleson and J. Charbonneau
Water continuous fat crystal networks in ice cream induced by unsaturated monoglycerides N. M. Barfod , Danisco Cultor, Brabrand, Denmark
High pressure application fo food systems and its impact on functional ingredients B. Tauscher*, P. Butz,and A.F. Garcia, Federal Research Center for Nutrition, Karlsruhe, Germany
Specificity and application of lipolytic enzymes in bread making processes T.B. Frandsen, T. Spendler, G. Budolfsen, L. Christiansen and J. B. Neilsen, Novozymes A/S, Bagsvaerd, Denmark
Afternoon Session 6: Colloidal and Interfacial Sciences - chairs D. Pechak and M. Paques
Rheology and Structure of Particulate Protein Gels T. van Vliet, Wageningen Centre for Food Sciences,Wageningen, The Netherlands
Posters:
1. In situ study of the effect of heating on dough components using Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR)
O. Sevenou, S.E. Hill, I.A. Farhat and J.R. Mitchell. Division of Food Sciences, University of Nottingham, Loughborough, UK
2. Antioxidative activity of the crude extract of lignan glycosides from sesame meal
Y-S Shue and L.S. Hwang, Graduate Institute of Food Science and Technology, National Taiwan University, Taiwan, R.O.C.
3. Near Field Microscopy of phase separation in a mixed interfacial protein/surfactant film
A. P. Gunning, A.R. Mackie, A.R. Kirby and V.J. Morris, Institute of Food Research, Norwich Research Park, Norwich, UK
Contact information for the chairs is shown below, in alphabetical order:
Paula Allan-Wojtas Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre, Kentville, Nova Scotia, Canada B4N 1J5 Tel: (902) 679-5566 FAX: (902) 679-2311 email: allanwojtasp-at-em.agr.ca
Judy Arnold USDA -ARS - RRC 950 College Station Rd. P.O. Box 5677 Athens, GA 30604-5677 USA Tel: (706) 546-3515 FAX: (706) 546-3068 email: jarnold-at-ars.usda.gov
Mark Auty Dairy Products Research Centre TEAGASC Moorepark, Femoy, Co. Cork Ireland Tel: 011-353-25-42447 FAX: 011-353-25-42340 email: mauty-at-moorepark.teagasc.ie
James E. Charbonneau National Food Processors Association Food Chemistry and Packaging Department 1401 New York Ave, NW Washington, D.C. 20005 USA Tel: (202) 639-5972 FAX: (202) 639-5991 email: jcharbo-at-nfpa-food.org
Kathy Groves Leatherhead Food Research Association Randalls Road, Leatherhead Surrey KT22 7RY England Tel: 44 0132 822330 FAX: 44 0132 386228 email: kgroves-at-lfra.co.uk
Tony McKenna New Zealand Dairy Institute Private Bag 11 029 Palmerston North, New Zealand Tel: 011 64 6 350 4649 FAX: 011 64 6 356 1476 email: tony.mckenna-at-nzdri.org.nz
Marcel Paques Wageningen Centre for Food Sciences/Unilever Research Vlaardingen P.O. Box 20, 6710 BA Ede The Netherlands Tel: 011 31 318 659690 FAX: 011-31-318 650400 email: paques-at-nizo.nl
David Pechak Kraft Technology Centre 801 Waukegan Road Glenview, IL 60025 USA Tel: (847) 646-4808 FAX: (847) 646-3864 email: dpechak-at-kraft.com
Delilah Wood USDA - ARS - WWRC 800 Buchanan Street Albany, CA 94710 USA Tel: (510) 559-5653 FAX: (510) 559-5777 email: wood-at-pw.usda.gov
Greetings, Does anyone have experience with lanthanum and/or ruthenium HT staining specifically for isolated cells?? I prepared two samples of single atrial cells, using protocols from the 60's and 70's. The ultrastructure was great, in part because I believe that the lanthanum as well as the ruthenium enhanced the UA and LC staining. I was hoping for a much more dense cell margin that would help to delineate vesicles that might be open to the surface. I know that Ruthenium HT is supposed to bind to the glycocalyx and stain well....but the lanthanum protocols I could find were all intact tissue where the spaces between cells were filled with dense material. I am not sure that lanthanum works for single cells, but gave it a try. Ruth. HT was added to both the GA and OsO4, but lanthamun was used, per protocol, only in the OsO4. Etoh dehydration and Embed 812 resin. Any sage wisdom on how to improve staining on a single cell prep would be helpful. Linda Fox lfox1-at-lumc.edu
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Fellow Microscopists: As many of you know, the Facility Management session held at M&M2000 was very well received and there was a unanimous request for it to be continued. Therefore a similar session will be held at M&M 2001. I must finalize the topics within the next week or two. Since the purpose of this session is to discuss topics of interest to facility managers, I would like your input before the session is finalized.
The topics covered in the M&M 2000 session with transcripts published in Microscopy Today were: 1) Multi-user Facilities...managing users 2) Justification of Costs, cost recovery, electronic bookkeeping and billing 3) Equipment Maintenance: Vender service contracts vs. Independent providers vs. Insurance company/self-insure options.
The following are some suggestions that were gleaned from the evaluation forms and discussion during the session.
1) Training Users…courses, workshops, …what works for whom? 2) Scientific Ethics…What are our responsibilities as lab Managers? 3) Mission statements/lab organization/lab business plans/future planning issues 4) Staffing issues including training, part-time student help, Training course T.A’s 5) Daily issues…prioritizing use, prioritizing service projects, dealing with difficult users/customers 6) Legal issues involving data ownership, confidentiality of data, handling of potential evidence. 7) Justification for new equipment 8) Commercial use of university facilities 9) Microscopy as a research tool…integration of industry or campus facilities in coordination with other core facilities. 10) Issues unique to Multi-user and/or Service facilities and how such facilities can coexist.
The first 2 received the greatest number of requests. However there may be more relevant topics now. Please rank the 3 of highest priority to you or suggest additional topics of interest. I will sift through all replies at the end of next week and narrow them down to 3. Also please feel free to supply the names of individuals to lead specific discussion topics. The format will be the same as the last session. A presenter will give a 10-15 min. introduction of the topic followed by ~30 min. open discussion.
Thanks, Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
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Hi All, On the "serious" side of this, and to make the safety police happy, EMS sells a plastic guard that holds razor blades, exposing just a small expanse for trimming. I had bought some years ago, on the "that sound good" premise. Personally, I found them awkward to use, but they are cheap, and having them in the lab (even if not used) may be enough to satisfy your safety wizards.
Disclaimer: I have no financial interests in EMS.
Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Mounting of particles on the order of 50-100um onto double stick tape is easily done as previously noted by using a very fine artist's brush and distilled water. But this should be done moist, rather than wet. After dipping the brush into DW, twirling the length of the brush end on a lab tissue both removes excess water and creates a finer point at the tip. The moist, fine-tipped brush can then not only pick up the diatom, foram or other particle, but can also be used to orient it and position it on the tape with good control. This method also prevents glue from bleeding up the particle. Particles of this size range can also be set down on a thin layer of silver conducting paint that's still wet, if you can chase the drying horizon fast enough, or on a thin layer of still-wet collodion (the same adhesive used by belly dancers to secure the navel jewel, but that's another story).
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Ed: Edmund Scientific sells test slides for microscopes. Perhaps you can find an adequate one there. Some test slides run up $300, though. Or, what about using diatoms to compare one scope with another?
Sam Purdy Technical Center/National Steel Corp Trenton MI } ---------- } From: ed_bachmann-at-unc.edu } Sent: Friday, January 2001, 11:01 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Nikon SMZ800 vs Leica MZ7 Stereozooms } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying to judge the optical quality of two similar stereoscopic zoom } scopes: the Nikon SMZ800 and the Leica MZ7, both equipped with a 1x } Planapochromatic objective. } } Leica publishes resolution figures for the MZ7, claiming 309 line pairs } per mm. } Nikon does not, and I haven't been able to persuade the Nikon } representative to } "characterize" the resolution of the SMZ800, although he says he can. I } do not } have the necessary test slide, nor do I know where I can get one with a } scale } graduated from say 200-400 lp/mm. I do know though that such a slide } would cost } probably $300-$500. } } Interestingly, Nikon publishes a resolution number for its next higher } quality } scope, the SMZ1000. It claims the SMZ1000 can resolve 300 lp/mm with a 1x } Planapo. Because Nikon product literature doesn't publish a resolution } for the } SMZ800, I suspect that its resolution is substantially lower than 300 } lp/mm, and } therefore inferior to the Leica MZ7. } } Does anyone have any suggestions on how I might judge the relative quality } of } these two scopes, or does anyone have experience with either scope that } might } shed light on this problem? Is my only alternative to find and buy a } suitable } test slide? If so, can anyone point me to a source? } } Thanks very much, } Ed } } Ed Bachmann } Odum Institute for Research in Social Science } Manning Hall CB 3355 } University of North Carolina at Chapel Hill } Chapel Hill, NC 27599-3355 } (919) 962-0512 } }
I believe that the time-honoured method of manipulating small particles with a fine brush can be improved on. A customer thought that certain specimen should not be wetted, because they were to be weighed. Presumably they could be left to dry, but that was inconvenient. I suggested one of those vacuum pick-up devices; these pick up specimens using a square tipped syringe needle. Obviously those needles are too large for foramifera but they can be tipped with thin plastic tubing. On the first try I pulled some 0.3mm internal tubing using a PE transfer pipette. With practice smaller tubes could be produced. I suggest that manipulating specimens with a vacuum pick-up is faster and more precise than using a brush. I have used a vacuum pick-up for placing (and turning over) TEM grids when making support films. Its a rather more elegant and faster method than using tweezers. Disclaimer: ProSciTech is a supplier of vacuum pick-ups. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, January 13, 2001 9:18 AM, Dee Breger [SMTP:micro-at-ldeo.columbia.edu] wrote: } } } Mounting of particles on the order of 50-100um onto double stick tape is } easily done as previously noted by using a very fine artist's brush and } distilled water. But this should be done moist, rather than wet. After } dipping the brush into DW, twirling the length of the brush end on a lab } tissue both removes excess water and creates a finer point at the tip. The } moist, fine-tipped brush can then not only pick up the diatom, foram or } other particle, but can also be used to orient it and position it on the } tape with good control. This method also prevents glue from bleeding up the } particle. Particles of this size range can also be set down on a thin } layer of silver conducting paint that's still wet, if you can chase the } drying horizon fast enough, or on a thin layer of still-wet collodion (the } same adhesive used by belly dancers to secure the navel jewel, but that's } another story). } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 914/365-8640 } F: 914/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) } }
Hi Listers, I've been thrown on the scrapheap at 60. So I'll be unsubscribing soon but will keep an eye on the list over a colleague's shoulder. Many thanks for all the help and interesting items, especially the forensic stuff - a fascinating and useful insight into the problems of others. Byeeee, Chris, Plant Path., IACR-Rothamsted, UK. chris.smith-at-bbsrc.ac.uk
Most injuries while trimming blocks with hand-held razor blades result either from the free hand not knowing where the razor hand is, or from sudden uncontrolled movement of the blade when force is released at end of cut, or due to slippage, etc. when the razor hand / arm movement is unlimited. A couple of simple rules will prevent these situations from arising. 1) Look before you move 2) when cutting, use both hands - hold one end of the blade in each hand at all times. That way you know where both hands are in relation to the blade all the time and there is also improved control over blade movement 3) Never cut with the blade at the end of an unsupported arm. Arms and hands should be supported as close to the specimen as possible, so that blade movement is strictly limited to the range that can be driven by deliberate finger movements, and free, involuntary movements cannot occur.
I recommend you not to wear gloves because they interfere with touch perception and interfere with your grip on the blade. Has your safety officer seen you in action? Does he really know what the risks are, or is his imagination running riot? Show him your procedures. (Any gender-specificity in the above is wholly unintentional)
Chris Jeffree University of Edinburgh Biological Sciences EM Facility
} Hi Listers, } I've been thrown on the scrapheap at 60. So I'll be unsubscribing soon but } will keep an eye on the list over a colleague's shoulder. Many thanks for } all the help and interesting items, especially the forensic stuff - a } fascinating and useful insight into the problems of others. } Byeeee, Chris, Plant Path., IACR-Rothamsted, UK. chris.smith-at-bbsrc.ac.uk
Chris -
WRONG attitude! I retired at the same age, and started Project MICRO so that I could continue to share my love of the microworld. I suggest that you contact the RMS equivalent, AMFES, and have some fun yourself!
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Dear Listers, Many thanks indeed for the unanticipated response to my 'for info' note. Attitude adjustment assimilated. The world will hear from me again! ttfn, Chris
Still looking...for a few good account representatives for an electron and optical microscopy service. territory is open., excellent earning potential. Please call Dick O'Connellat 734-668-3309 or e-mail Oconnell-at-ltu.edu. for more information. (I was unable to reach some of you who responded because of one reason or another--you know who you are. Please contact me again with a phone number or some other means to reach you.)
We are looking for someone with EDS/SEM experience to work out of our Florida office. Responsible for setting up procedures and repairing systems in house as well as customer sites. Working without supervision is a must. if you are interested to know more about the position, please send your qualification and salary history to our Email address. rangets-at-aol.com
Dear Colleage, Does anybody know if there is a software that can create the atomic structure for an interface or a dislocation, so that an image simulation (by using EMS or Mac Tempas) can be carried out directly for such structures? I know crystal Kit can create an interface structure,however, its atomic cordinations can not be exported directly to MacTempas or EMS directly for image simulations.
I would be greatly appreciate if you could provide such kind of information!
Sincerely,
James
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Hello, I a trying to stain sections of plant root that has been infected with a fungus to highlight certain features including lignin, callose, suberin and the fungal tissue itself. However, my samples have been embedded in LR White and Spurr's Resins. I am having difficulty finding protocols for staining of these resins. Most protocols refer to paraffin sections and it is not possible to use ethanol based stains for these resin sections. Does anyone know of any good references, books etc where I can find protocols for staining of these resins for LM? Thanks for your help. Rose
Several stains are used with resin embedded sections. The favorite seems to be Toluidine blue. This topic was discussed last March and several postings will be in the Archives. The differentiation method I contributed then, but to make this posting useful, I'll give here "my" complete method. There are many variants, most recently contributors to the histoserver were "frothing" how wonderful the results of staining with a combination of urea and Toluidine blue are - I have no experience with that.
Make up 0.5% Toluidine blue and 1% Borax in water. This will keep "forever" in a stoppered bottle on the bench. Keep with it a small funnel and a double layer filter paper.
Sections should be well adhering to a slide after heating on a 60-80 degree hotplate for some minutes after the sections had dried. Run a thick felt-tip pen around the section - on the underside. This will help locating sections and will not wash off.
Poor about 5ml of stain into the filter and apply a couple of drops to the sections. Sit the funnel back on the bottle recovering the stain.
Place the section on a hotplate and leave until the edge of the stain has dried up.
Now a few drops of water could be added to gently remove the remaining stain. This would result in a fairly intense, but blue-only staining.
Its better to differentiate. This is one of the most attractive features of the Toluidine stain as it can give a two colour, pink and blue rendition. This shows cell features similar to a double staining with Haematoxylin & Eosin.
After staining on a hotplate add a drop of acid alcohol and then rinse the slide with distilled water. The acid alcohol can destain partially or even completely, but a brief application brings forth the two colours.
Make acid alcohol by adding to 25ml of 50% Ethanol a couple of drops of 1N HCl. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, January 16, 2001 1:59 PM, Rosalie Daniel [SMTP:rosalie-at-deakin.edu.au] wrote: } } Hello, } I a trying to stain sections of plant root that has been infected with a } fungus to highlight certain features including lignin, callose, suberin and } the fungal tissue itself. However, my samples have been embedded in LR } White and Spurr's Resins. I am having difficulty finding protocols for } staining of these resins. Most protocols refer to paraffin sections and it } is not possible to use ethanol based stains for these resin sections. Does } anyone know of any good references, books etc where I can find protocols } for staining of these resins for LM? } Thanks for your help. } Rose } }
LR White is a methacrylate, and most routine paraffin stains can be used directly with minor time modifications (the methacrylate can give some background staining, but usually umimportant). Spurr's (and other epoxies) can also be stained with differential methods--primarily using Toluidine Blue or methylene blue. Check out Hayat's book on staining (MA Hayat, "Stains and Cytochemical Methods", Plenum Press, 1993). He includes a broad range of stains, including PAS, basic fuchsin-toluidine blue (similar to H&E), Azure II-methylene blue-safranin O, H&E, and a thionin-acridine orange (specifically for plant tissues). Another good source of references would be to go to PubMed (http://www.ncbi.nlm.nih.gov/PubMed) and do a search for stains, then for resins or methacrylate, combine your searches, and use the original papers (I know there are a lot out there). BTW, most suppliers of stains (e.g. EM Sciences, Polysciences, Ted Pella, EB Sciences, Fullam, Ladd, ProSciTech,SPI, etc) will have protocols for using their stains on LR White and Spurr's--another good source for you. Hope this helps.
Roger C. Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
I have no financial interest in any of the mentioned vendors/suppliers (and if I missed anyone--I apologize, it was purely a senior moment).
(On Tue, 16 Jan 2001 14:59:03 +1100, Rosalie Daniel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I a trying to stain sections of plant root that has been infected with a } fungus to highlight certain features including lignin, callose, suberin and } the fungal tissue itself. However, my samples have been embedded in LR } White and Spurr's Resins. I am having difficulty finding protocols for } staining of these resins. Most protocols refer to paraffin sections and it } is not possible to use ethanol based stains for these resin sections. Does } anyone know of any good references, books etc where I can find protocols } for staining of these resins for LM? } Thanks for your help. } Rose } } }
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Hello, I work with radicle tips and I'm looking for an effective stain in order to reveal different cellular types such as xylem and/ or phloem, but I'm havin some problemsa in finding protocols. Can anybody help me? Thanks Carmen Martín
One of our students was worried and put labeling tape (sometimes referred to as "time tape") along the edge to be held (we normally break a double edge in half). We also have a very graphic cartoon drawn by our grad student depicting a severed finger complete with blood (he was getting frustrated with the EM students...)
Regarding the safety officers - teach *them* to section...
John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
Hi Linda, I worked with isolated ventricular cells many years ago, and as I remember, there was always some variablilty of staining intensities between cells that were processed in the same sample. We used cell that were isolated using enzymatic treatment in conjunction with shaking ( I believe that was pretty standard. This was in the early 1980's, work with Beatrice Wittenberg and Thomas Robinson). Cells that ended up in the same dish for study, and were fixed & processed together, took the stains differently. We also used Ruthenium Red to stain the glycocalyx as well as Alcian Blue, Safraninin O, phenylene diamine and a few other stains. You can look up the papers by Simionescu & Simionescu from the 1970's for examples. Caveoli can also be visualized with careful staining of silver-grey sections. You can also try bismuth instead of lead, but use it at 1/10 the recipe's strength and for a shorter time.
good luck, Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
The Department of Biological Sciences at Chicago State University is looking for an experienced electron microscopy technologist. We teach a course in TEM in the fall and SEM in the spring. In the summer we have students from several programs who learn about EM and complete a simple research project. We work on research projects with undergraduates, graduate students, and
faculty. We have a really nice facility with JEOL 1200 STEM, JEOL 2100 SEM, and RMC ultramicrotomes with cryo. We have a beautiful campus that is very easy to get to as it is right at
several major freeways. Our website will give you more information: WWW.CSU.EDU/jbrown/EM
In our lab we have been sputter coating with gold since the dawn of time. This has essentially been the "traditional" metal we have used for coating our non-conductive samples. We have found that in some of our applications that the gold grain is to large and is causing us problems in visualizing some of the smaller structures (primarily in engineering samples) that we are interested in. We currently sputter our samples for 1 to 4 minutes with a current reading of 18 to 20 mA using an argon atmosphere in the coater. We are aware that chromium and/or osmium coating are preferential to gold sputtering, but those technologies are not available to us at this time. What I would like to know is: 1) What are the advantages of using something other than gold as the target material (eg. Gold palladium, platinum palladium, or just platinum)? 2) Are there other materials that will work besides these metals that would give us equal service for small grain and a stable coating? 3) Are there operating conditions we could try that would still give us good coating but reduce the grain size of the film material? Thanks for your help,
Mike
====================================== Michael D. Standing BYU Microscopy Lab 401 WIDB Brigham Young University Provo, UT 84602
Hello listers- I am looking for a supplier of bakelite ring forms 1 inch in diameter (outside). Would someone contact me with that information. I can't find anything on the web. I appreciate your help! Many Thanks, Sarah -- Sarah A.W. Lundberg Electron Microanalysis and Imaging Laboratory Department of Geoscience, UNLV 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010
Dear Michael, you dont say what resolution you need, but if you cut the current on your existing gold system down to 4-5mA for 3 min or so, you may see a considerable improvement in the 10-50,000X range.
Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU EMU Box 475 Canberra ACT 2601 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 or 6125 8525 http://www.anu.edu.au/EMU
} } } "Michael D. Standing" {Michael_Standing-at-byu.edu} 01/17/01 07:37am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers:
In our lab we have been sputter coating with gold since the dawn of time. This has essentially been the "traditional" metal we have used for coating our non-conductive samples. We have found that in some of our applications that the gold grain is to large and is causing us problems in visualizing some of the smaller structures (primarily in engineering samples) that we are interested in. We currently sputter our samples for 1 to 4 minutes with a current reading of 18 to 20 mA using an argon atmosphere in the coater. We are aware that chromium and/or osmium coating are preferential to gold sputtering, but those technologies are not available to us at this time. What I would like to know is: 1) What are the advantages of using something other than gold as the target material (eg. Gold palladium, platinum palladium, or just platinum)? 2) Are there other materials that will work besides these metals that would give us equal service for small grain and a stable coating? 3) Are there operating conditions we could try that would still give us good coating but reduce the grain size of the film material? Thanks for your help,
Mike
====================================== Michael D. Standing BYU Microscopy Lab 401 WIDB Brigham Young University Provo, UT 84602
Ah, once again invaluable information, Thank you John, Fred and Nestor too!! I tried all the suppliers I could think of, except Buehler of course!
-- Sarah A.W. Lundberg Electron Microanalysis and Imaging Laboratory Department of Geoscience, UNLV 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010
Hi All, Here's an LM-related question: A group here needs to look at samples stained with picrosirius red using polarizing optics. I have a microscope equipped with DIC, and realize that if we pull the Wollaston prisms, we might be able to see something, but I don't have a rotatable stage which would make it easier to align their samples relative to the polarized light. Do any of you out there in the New York City area have an appropriate microscope? Or any ideas? TIA, Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
We at South Bay Technology offer such a product. They are available as our Part Number RFP100-10 and cost $5.25 for a package of 10. We have them in stock and can ship them out the same day you order them. I hope this helps.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Sarah Lundberg } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello listers- I am looking for a supplier of bakelite ring forms 1 inch in diameter (outside). Would someone contact me with that information. I can't find anything on the web. I appreciate your help! Many Thanks, Sarah -- Sarah A.W. Lundberg Electron Microanalysis and Imaging Laboratory Department of Geoscience, UNLV 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010
Hi all, I am working with seeds of Ilex paraguariensis (the maté). The aim of my research is to describe the ultrastructure (TEM) of the endosperm and the imature embryo. The samples (small dissected pieces) were fixed in glutaraldehyde and formaldehyde solution in phosphate buffer, washed, transferred for osmium (2% in 0,8% K3FeCN6, 1:1), dehydrated in acetone (10, 30, 50, 70, 90, 90 ,100, 100, 30 min in each step) and included in 1:3, 1:1, 3:1 and two changes of pure Spurr resin (24 hour in each step with continuous agitation). However, the tissues are very poorly infiltrated (endosperm and embryo with holes and soft parts). The endosperm cells have a lot of proteins and lipids as storage substances. I think that I should use a special protocol for this plant tissues, but I want to maintain the osmium fixation (lipid preservation). I tried the LR White and Unicryl resins with worst results. Does anybody suggest some thing? Thank's.
Dr. Rinaldo Pires dos Santos Lab. of Plant Anatomy Dept. of Botany - UFRGS Brazil e-mail: rinaldop-at-uol.com.br
you will find a lot of information in "Histochemistry" by R.W. Horobin, Stuttgart, G. Fischer 1982.
A few years ago I did some staining of plant cuticles (lets assume cutin is similar to suberin for your purpose) with Sudan Black B in Epon sections. SBB needs to be dissolved in hot alcohol but it worked quiet well on my Epon semis. I even managed to dissolve the resin in KOH before staining and got good results ... A recipe for SBB staining you will find in Bronner, R., 1975: Simultanous Staining of Lipid and Starch in Plant Tissues. Stain Technology 50(1):1-4.
I do not know if you want to use Toluidin Blue O, but TBO is a water based stain for acid polyanions like pectins, DNA/RNA and stains cytoplasm too. It will stain paraffin and resin sections and is easy and relatively safe to use. A very basic paper describing it is: Sakai, S.W, 1973: Simple method for differential staining of paraffin embedded plant material using Toluidin Blue O. Stain Tech 48(5):247-249.
Hope that helps you,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
------- Forwarded message follows ------- } From: Alexander Tikhonovski {tikhonov-at-mpi-halle.mpg.de} To: "Pier, Julie (LNA)" {Julie.Pier-at-america.luzenac.com}
Hi Rinaldo,
I have been working on the ultrastructure of orchid stigmas for a few years, this tissue contains a large amount of lipids and intercellular cutin, debris from plasmolysed "nutritive cells", slimy pectins etc ... very difficult to embed - at least that was what everbody told me when I started. As in the orchid column the pollen (pollinia in this case) is still there, in most cases I embedded even the pollen with its sporopollenin, a lipidic substance usually extremely bad to embed (sections may break out at the interface to the resin).
In the beginning I tried Spurr and had very poor results. Then I switched to Epon although everybody told me that this would work even worse because of the high viscosity of Epon (= bad infiltration).
But I could achieve godd embedding with Epon for my tissue. Here is how I did it:
Specimen: Orchid buds, columns and stigmas, preferably smaller then 2x2x2 mm Fixation: 3% GAD in cacodylate buffer pH7.2 for at least 24 h Wash a few times in buffer and then in a.d. Stepwise dehydration in acetone: 30, 50, 70, 90, 100, 100% for 20 min each Intermedium acetontril 3 x 100% for 10 min each Embedding in Epon (Agar 100 Resin Kit): No movement of the specimens needed, just use Eppendorf tubes or something similar acetonitril:Epon 3:1 overnight acetonitril:Epon 1:1 4 h acetonitril:Epon 1:3 4 h pure Epon overnight change to Epon + accelerator Polymerisation in flat embedding form 48 h at 60°C
I used the acetonitril as a substitute for propylene oxide simply because it is said to be less poisonous. Maybe its the acetonitril step that is responsible for the good infiltration - I really do not know, but it worked well, so "never change a winning team".
Hope that helps you,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
15th International Congress on Electron Microscopy
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1 - 6 September 2002
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Mike, The obvious grains of Au are the reason that the Au/Pd alloy is used. It should be less apparent with a Au/Pd target. Ken Converse owner Quality Images Delta, PA
Michael D. Standing wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers: } } In our lab we have been sputter coating with gold since the dawn of time. } This has essentially been the "traditional" metal we have used for coating } our non-conductive samples. We have found that in some of our applications } that the gold grain is to large and is causing us problems in visualizing } some of the smaller structures (primarily in engineering samples) that we } are interested in. We currently sputter our samples for 1 to 4 minutes with } a current reading of 18 to 20 mA using an argon atmosphere in the coater. } We are aware that chromium and/or osmium coating are preferential to gold } sputtering, but those technologies are not available to us at this time. } What I would like to know is: } 1) What are the advantages of using something other than gold as the target } material (eg. Gold palladium, platinum palladium, or just platinum)? } 2) Are there other materials that will work besides these metals that would } give us equal service for small grain and a stable coating? } 3) Are there operating conditions we could try that would still give us good } coating but reduce the grain size of the film material? } Thanks for your help, } } Mike } } ====================================== } Michael D. Standing } BYU Microscopy Lab } 401 WIDB } Brigham Young University } Provo, UT 84602 } } e-mail: Michael_Standing-at-byu.edu } phone: (801) 378-4011 } fax: (801) 378-3937 } ====================================== } } } } }
I would suggest you try gold/palladium. It has a smaller grain size and the same reflectivity.
John Arnott
Disclaimer: Ladd Research sell targets for most types of sputter coaters. --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
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Michael D. Standing wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Dear Listers: } } In our lab we have been sputter coating with gold since the dawn of time. } This has essentially been the "traditional" metal we have used for coating } our non-conductive samples. We have found that in some of our applications } that the gold grain is to large and is causing us problems in visualizing } some of the smaller structures (primarily in engineering samples) that we } are interested in. We currently sputter our samples for 1 to 4 minutes with } a current reading of 18 to 20 mA using an argon atmosphere in the coater. } We are aware that chromium and/or osmium coating are preferential to gold } sputtering, but those technologies are not available to us at this time. } What I would like to know is: } 1) What are the advantages of using something other than gold as the target } material (eg. Gold palladium, platinum palladium, or just platinum)? } 2) Are there other materials that will work besides these metals that would } give us equal service for small grain and a stable coating? } 3) Are there operating conditions we could try that would still give us good } coating but reduce the grain size of the film material? } Thanks for your help, } } Mike } } ====================================== } Michael D. Standing } BYU Microscopy Lab } 401 WIDB } Brigham Young University } Provo, UT 84602 } } e-mail: Michael_Standing-at-byu.edu } phone: (801) 378-4011 } fax: (801) 378-3937 } ======================================
I have been doing cryoultramicrotomy for the last year or so with varying levels of success. I finally got some pretty pictures, so I know I can get it right. Now I want to make it consistent.
So, my biggest problem appears to be with freezing. I am fixing my cells with 4% para, 0.1% glut, 0.25M Hepes, rinsing with Hepes, embedding in 10% gelatin and then infiltrating with 2.3M sucrose in a stepwise manner. I am cutting the sample into *very* small blocks (less than 0.5mm) hoping to infiltrate better. I then place the sample onto pins and drop into liquid nitrogen to freeze. However, it always seems that only the very outer layer of tissue/cells are well frozen. If I take a 1 micron section to see how the tissue looks, I can't take another b/c it would bring me into a freeze damaged area. I have tried using liquid proprane and liquid freon, but the samples cracked when I attempted to section them.
I have now been given a very precious human sample that they want to immunolabel on cryo thin sections, and I don't have enough tissue to make lots of blocks to section just the outside.
If anyone has any suggestions to help me get good freezing deeper in the tissue, I'd appreciate it. TIA Leslie
These are my pretty pictures: http://www.aecom.yu.edu/aif/gallery/TEM/shields_immuno/tem_cryo.htm Analytical Imaging Facility visit our web site: www.aecom.yu.edu/aif Albert Einstein C. of M 1300 Morris Park Ave Forchheimer 639 Bronx, NY 10461 718-430-3547
It always amazes me how much discussion can be generated on some topics!
I very rarely offer my 'two cents', but love reading through the comments. On the issue of microtome safety: I have been an electron microscopist for over 25 years (I started as a child prodigy!) and other than one incident early on when I (stupidly) tried trimming a block by bringing the razor blade toward me, I have NEVER had a problem with severed fingers, major lacerations, etc.
The aforementioned incident did result in stitches and I bear the scar proudly.
I do my trimming on a special holder on the Reichert Ultracut looking through the binoculars and "map" out the block face by marking the edges with the blade. Then I proceed very carefully (away from me and the block) to trim the epon. I then do the rough sectioning on the microtome itself. At times, we have used a jeweler's saw and vise to trim away excess epon and get down to the tissue, etc.
I agree that gloves or other enhancements only add to the problem.
I believe the main thing to consider is: "Think before you cut!"
Peggy Sherwood -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The sleeves need to be 3 1/4 x 4 in. I don't know where they were bought } previously and need help finding out where. } } Thanks } } Cindy K. } } Joyce L. Kleier } Northwestern University, Evanston, IL. USA } j-kleier-at-northwestern.edu
Polysciences in Warrington, PA 800-523-2575 Electron Microscopy Sciences in Ft. Washington, PA 800-523-5874
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Hennepin County Medical Center, Minneapolis, MN has a used ISI-60A SEM available to anyone interested. Supporting equipment also available include: Eiko Multicoater, model VX-10A and an OMAR critical point dryer, model SPC 900/EX.
With emphasis on Tripod Polishing and Focused Ion Beam Specimen Preparation
.will be offered in conjunction with Surface Analysis 2001 and the Joint Annual Meetings of the Florida Society for Microscopy and the Florida American Vacuum Society in Orlando at the University of Central Florida March 14-16, 2001.
Instructors:
Ron Anderson, IBM Fred Stevie, Lucent Technologies Lucille Giannuzzi, UCF
Vendor Sponsors will be participating!
To register, please go to www.dce.ucf.edu
For technical information please contact Lucille Giannuzzi at lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Group, One of my readers has asked the following question. If you can help, kindly contact her direct. Don Grimes, Microscopy Today
Do you know or can you ask your readers about the possibility of Electrostatic Discharge damage caused by an SEM? Is this a problem? My company is reluctant to have me examine flight-ready devices in the SEM because of this possibility. I can't find any information on the subject. Thanks for any help you can give me.
Well, one of the (supposedly true)legends in our facility is that a student was knocked out of a chair one day long ago by a major spark from an old JSM35 SEM. But maybe that's not what you meant.... ;-)
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-8304 (573) 884-5414 (fax) email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu} http://biotech.missouri.edu/emc
-----Original Message----- } From: Don Grimes [mailto:microtoday-at-mindspring.com] Sent: Wednesday, January 17, 2001 3:23 PM To: microscopy-at-sparc5.microscopy.com
Group, One of my readers has asked the following question. If you can help, kindly contact her direct. Don Grimes, Microscopy Today
Do you know or can you ask your readers about the possibility of Electrostatic Discharge damage caused by an SEM? Is this a problem? My company is reluctant to have me examine flight-ready devices in the SEM because of this possibility. I can't find any information on the subject. Thanks for any help you can give me.
Does anyone know where to find a Teflon stirrer/mixer attachment in the shape of a corkscrew. We used to use one at the New Mexico State EM lab (yup, that's you, Soumitra) for mixing resins, attached to a variable speed drill-type mixer on a stand. I have never been able to locate another and my stirring arm is about worn out.
Thanks
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Dear Colleage, Does anybody know if there is a software that can draw the atom arrangments in a specific crystalline plane? I know several software, such as Crystalkit, Mactempas and Crystal Maker, can draw the atomic projection, in which many layers are overlapped, however they can not display the atomic arrangments in one specific plane.
I would greatly appreciate if you could provide such kind of information!
Sincerely,
James
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com
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Reply to: RE: Microtome Safety I too have been surprized by the response to this subject. I wonder if everyone who uses Lowicryl resin also hand trims the blocks with a blade?
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
} } To all: } } It always amazes me how much discussion can be generated on some topics! } } I very rarely offer my 'two cents', but love reading through the comments. } On the issue of microtome safety: I have been an electron microscopist for } over 25 years (I started as a child prodigy!) and other than one incident early } on when I (stupidly) tried trimming a block by bringing the razor blade toward } me, I have NEVER had a problem with severed fingers, major lacerations, etc. } } The aforementioned incident did result in stitches and I bear the scar proudly. } } I do my trimming on a special holder on the Reichert Ultracut looking } through the binoculars and "map" out the block face by marking the } edges with the blade. Then I proceed very carefully (away from me } and the block) to trim the epon. I then do the rough sectioning on } the microtome itself. At times, we have used a jeweler's saw and } vise to trim away excess epon and get down to the tissue, etc. } } I agree that gloves or other enhancements only add to the problem. } } I believe the main thing to consider is: "Think before you cut!" } } Peggy Sherwood } -- } Peggy Sherwood } Lab Associate, Photopathology } Wellman Laboratories of Photomedicine (W224) } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-724-4839 (voice mail) } 617-726-6983 (lab) } 617-726-3192 (fax) } sherwood-at-helix.mgh.harvard.edu } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A58458C70244; Wed, 17 Jan 2001 16:15:00 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA15685 } for dist-Microscopy; Wed, 17 Jan 2001 11:00:39 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA15682 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 17 Jan 2001 } 11:00:09 -0600 (CST) } Received: from helix.mgh.harvard.edu (helix.mgh.harvard.edu [132.183.108.14]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id KAA15672 } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Jan 2001 10:59:58 -0600 (CST) } Received: from [132.183.150.185] by helix.mgh.harvard.edu } (8.8.8/1.1.20.3/31Dec98-1151AM) } id LAA0000025503; Wed, 17 Jan 2001 11:52:15 -0500 (EST) } Mime-Version: 1.0 } Message-Id: {a05001902b68b7bb661c5-at-[132.183.150.185]} } Date: Wed, 17 Jan 2001 11:55:27 -0500 } To: Microscopy-at-sparc5.microscopy.com } From: Peggy Sherwood {sherwood-at-helix.mgh.harvard.edu} } Subject: Microtome Safety } Content-Type: text/plain; charset="us-ascii" ; format="flowed" } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273279032 } Status: U }
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id TAA17086 for dist-Microscopy; Wed, 17 Jan 2001 19:53:12 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id TAA17083 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 17 Jan 2001 19:52:42 -0600 (CST) Received: from hall.mail.mindspring.net (hall.mail.mindspring.net [207.69.200.60]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id TAA17076 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Jan 2001 19:52:31 -0600 (CST) Received: from mindspring.com (user-2ivemhk.dialup.mindspring.com [165.247.90.52]) by hall.mail.mindspring.net (8.9.3/8.8.5) with ESMTP id UAA01008 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 17 Jan 2001 20:50:56 -0500 (EST) Message-ID: {3A664C2C.8E2581AB-at-mindspring.com}
James,
Perhaps you are familiar with an older version but CrystalMaker can limit the size of your structure to one atomic plane and draw an opaque or translucent plane in any color parallel or coincident ot the plane of interest. Check it out.
{http://www.crystalmaker.co.uk}
Boron nitride wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Colleage, } Does anybody know if there is a software that can draw the atom arrangments } in a specific crystalline plane? I know several software, such as } Crystalkit, Mactempas and Crystal Maker, can draw the atomic } projection, in which many layers are overlapped, however they can not } display the } atomic arrangments in one specific plane. } } I would greatly appreciate if you could provide such kind of } information! } } Sincerely, } } James } } _________________________________________________________________ } Get your FREE download of MSN Explorer at http://explorer.msn.com
-- Gordon Nord Scientist Emeritus US Geological Survey Email gnord-at-mindspring.com
I think you are doing everything right (fixation, rinsing, 10% gelatin, 0.5mm size, drop into LN2), but maybe it is the STEPWISE infiltration with 2.3 M sucrose that does not work well for your cells. How long do you do the infiltration? At what temperature?
You probably get a gradient of sucrose concentration from the edges to the center of your blocks because the lower suc concentrations enter the block rapidly while the higher concentrations need more time?! The different concentrations probably do not have time enough at low temperatures (i.e. +4°C) to mix/substitute properly.
I would recommend to do the infiltration in an Eppendorf vial (app. 1ml) DIRECTLY in 2.3M sucrose at +4°C (your gelatin is not fixed I assume, so you need the low temperature for the gelatin to stay solid). Infiltrate for AT LEAST 2 HOURS on a rotating table.
Jan Slots group in Utrecht, The Netherlands achieves very good results with that method. They are giving a highly recommendable CRYOCOURSE from 3rd - 12th July 2001 in cooperation with Leica (I am doing a tiny little bit of it). If you are interested I can send you detailed information.
Hope that helps you,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Your question is quite frankly hard to fathom. In an SEM, the sample is subjected to up to 50 KV electron bombardment intentionally at up to several hundred nanoamps in an area as small as a few tenths of a square millimeter. Exactly what measure are you considering as electrostatic discharge? Voltage, current or radiation flux density? And what specs are you holding your electronics to? If lightning strikes or EMP effects are considered, the SEM's effects are probably minimal.
50 KV can certainly be considered an electrostatic potential. 50 KV at 100 nanoamps sample current translates to only 5 milliwatts. However, that amount of power translated to the sub-millimeter scales of an electron microscope can add up, as can the effects mentioned when considering such small structures.
A rough estimate shows that a 30 KV beam at 100 nA sample current and an excitation surface on the sample of 100 micrometers diameter yields a huge energy density at the surface. Assuming a subsurface absorption sphere of around 200 micrometers diameter, obviously the radiation flux densities are quite high. I leave it to you to calculate the actual figures based on the materials you are sampling.
However, given your internet domain and the specific question asked, I have to wonder what reservations your company has. If there are electrostatic weaknesses in the circuitry you want to examine, is it better to have these brought out in R&D SEM examination or in field exposure to lightning, EMP and ECM? If I were in the left seat, I would want some assurance that those envelope edges had been explored. My intuitive side would tend to believe that the potential lightning, EMP and ECM effects would likely be as concentrated, if not more so, that the SEM beam energies in the nanoscale structures of the circuitry you are probably using. In fact, the electron beam energies may be the best method you have of simulating the localized energy density effects of these sources.
} Group, } One of my readers has asked the following question. If you can help, kindly } contact her direct. } Don Grimes, Microscopy Today } } Do you know or can you ask your readers about the possibility of } Electrostatic Discharge damage caused by an SEM? Is this a problem? My } company is reluctant to have me examine flight-ready devices in the SEM } because of this possibility. I can't find any information on the } subject. Thanks for any help you can give me. } } Virginia Harper } vbharper-at-west.raytheon.com } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
We get them from Better Containers Mfg. Co. 530 Hyde Park Hillside, Ill 60162 708-547-7272
There is also other companies (e.g. Consolidate Plastics Co., Inc. 330-425-3900) out there.
Good Luck
On 17 Jan 2001, at 10:10, Cindy Kleier wrote:
} The sleeves need to be 3 1/4 x 4 in. I don't know where they were } bought previously and need help finding out where. } } Thanks } } Cindy K. } } Joyce L. Kleier } Northwestern University, Evanston, IL. USA } j-kleier-at-northwestern.edu }
John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
} } The sleeves need to be 3 1/4 x 4 in. I don't know where they were bought } previously and need help finding out where.
We carry Plastine sleeves for 3 1/4 x 4 negatives. These sleeves are archival quality polyethylene and come in a box of 1000. We also carry Kodak Sleeves which come in boxes of 100, and the Print File EM-6 page which will hold 6 negatives in a page that fits a 3 ring binder.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
We are preparing Pt-C and C replicas from different biological material. For cleaning the obtained replica we either use bleach (5.2%) or chromic acid (10, 20 followed by 40%) with washes in between. The protocol usually works well for most stuff but when it comes to oocytes from Xenopus we are usually left with a lot of organic material still attached to the replica. If we leave the replica in cleaning agent for a long time we end up with a very brittle replica which can easily break.
Oocytes have got a reputation for being very sticky and difficult to clean. We are considering to use plasma cleaning instead but I don't know what to expect from that. I would appreciate it if you could share your 'tips and tricks' with us.
Majid Ghoddusi
................................................................ Majid Ghoddusi, PhD Protein Dynamics Unit Department of Chemistry Australian National University ACTON 0200 Canberra Tel: (02) 6125 4190 Fax: (02) 6125 0760 e.mail: majid.ghoddusi-at-anu.edu.au web site: http://langevin.anu.edu.au/ ........................................................................... ..........
Hi Leslie, One thing I can think of, is that you shouldn't drop your samples into the liq. Nit. LN2 is a poor cryogen in that it boils when warm things are placed into it, creating a pocket of nitrogen gas around the sample, that acts as an insulating layer. Most of us do use nitrogen to freeze our samples. I always hold the pin end of the stub in a hemostat-type clamp and wiggle it as it freezes to try to ensure an exchange of the cryogen and avoid too much of a gas bubble around the sample. I'm sure you will get many other suggestions, since there are so many of us out there doing this with varying degrees of success. Good luck, Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
ESD damage on semiconductor samples is quite real & has been documented for years. When CMOS circuits first came out this was a real concern. Motorola & a number of other semiconductor companies has reams of papers on this subject.
Contact me offline & I can see if I can dig up the info.
Regards,
Earl
----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Don Grimes'" {microtoday-at-mindspring.com} ; {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 18, 2001 2:10 AM
Hi
Just a few points which may help in your quest for better sputter coatings.
Sputter coaters fall into a number of categories based upon age. Each category requires a slightly different set up procedure.
Class 1 - Coaters with a variac voltage control of up to 1500volts Class 2 - Coaters with a variac voltage control of up to 2500volts Class 3 - Cool coaters with a fixed voltage but a variable "deposition" control
General - All coaters need approximately a 5cm target to specimen distance. Coat a piece of paper the 2-3 minutes the paper should cover the specimen bed, you will then see the distribution pattern of low and high depostion areas. Make sure you put your specimen in a high deposition area. Coating many specimens in one run may lead to uneven coatings as each specimen has an affect on those near by, outgassing, partial shadowing etc.
Class 1 need an operating voltage which you would guess is about 700 volts on the voltage dial, Class 2 around 800 volts (i.e. as low as possible whilst still striking a plasma). These coaters probably need a 1 minute coating at 20mA for gold.
Class 3 coaters need a deposition rate set at 10mA and a 30 second coating for gold.
With any coater increase the coating time and you increase thickness and the possibility of visualizing the structure.
Multi coats increase the visibility of the metal structure.
Platinum deposits at half the rate but at less than half the grain size; increase coating times but do not exceed the 20mA and 10mA settings as described above.
Hope this helps
Steve Chapman Senior Consultant Protrain For consultancy and training by professionals World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
Thanks to all of you who have responded to my query about polarizing microscopes. Among the responses was the name of someone nearby who has a fully equipped instrument, and if all the "poor man's" adaptations I've received don't pan out, I'll give that fellow a call. Below are highlights of the responses I received. We can put this topic to rest now.
Lee
*********************** Hello re: rotating stage.
Put some vaseline on top of your stage after removing the mechanical stage, and invert an AOL CD on it. Makes an elegant rotating stage for a scope. ************************ Alternatively, you can make a mini "Theta" stage by mounting any sort of small turntable on your existing stage. Take off all the normal slide holding/mechanical stage equipment and mount the turntable in place. Parameters: a. You need to open a hole in the center so that you can get light through the slide. b. You need some mechanism (as simple as a rubber band or an old stage clip) which will allow you to slide the slide for XY; the special stage will supply the rotation c. Depending on the thickness of the turntable, you may need a long working distance condenser; simple Abbe condensers are good, but for true pol work, you need high NA
There are limits to this approach, especially in terms of centration, but if they are just doing qualitative Pol, it's a good way to go. *********************** Yes, as you wrote, if you remove the wallaston's from the light path, you will indeed have a set up with crossed polarizers. This is a good start, but as you suggested that still leaves a few problems. You mentioned the rotating stage. As DIC is fairly popular, you might well find someone around with a DIC stand that has a rotatable stage. Really, for DIC work, the stage ought to rotate too, because contrast in the specimen does change as a function of the orientation of the sample. But there are two other very useful things that you should look for in the stand.
1) Be sure that you can rotate either the polarizer or the analyser. For DIC, it doesn't matter all that much if the analyser and polarizer are not exactly crossed. But for polarized light they need to be crossed as exactly as possible. You can look through the eyepieces at a blank slide and rotate either the analyser or the polarizer until the field gets maximally dark. Then stop.
2) A compensator of some kind is handy. That is because often the sample has weak retardation and the compensator enhances that. If you are lucky enough to find a real polarized light scope, it will have several of these available. But if you use a DIC scope, then what you can do is to experiment with various kinds of plastic that you find lying around. Petri dish lids are a good bet, or clear tape or plastic wrap stretched on a slide. Put this in the beam, somewhere after the polarizer in a way that you can rotate to some extent. You'll know you have got the right thing when you find something that makes the nice dark field (from step 1) get bright as your rotate it. *********************** I have made a 'poor-man's' polarizing microscope by putting one sheet of polaroid material (glass or plastic) over the light source of an ordinary LM and another sheet between the slide and the objective lens. I rotate the sheet over the light source until I get the desired effect. Works great as long as you do not need a high magnification lens that works close to the specimen.
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
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Okey, one last time. Please, if you haven't weighed in on your preference for topis (see below) by tomorrow, I will pick them based on response to date. There is a wide range of responses and I would love to have more to see if a few topics end up standing above the rest as to interest level. Also, don't forget to recommend presenters for specific topics. And thanks to all who have already responded. Please don't stuff the ballot box by voting again. Thanks, Debby
=================
Fellow Microscopists: As many of you know, the Facility Management session held at M&M2000 was very well received and there was a unanimous request for it to be continued. Therefore a similar session will be held at M&M 2001. I must finalize the topics within the next week or two. Since the purpose of this session is to discuss topics of interest to facility managers, I would like your input before the session is finalized.
The topics covered in the M&M 2000 session with transcripts published in Microscopy Today were: 1) Multi-user Facilities...managing users 2) Justification of Costs, cost recovery, electronic bookkeeping and billing 3) Equipment Maintenance: Vender service contracts vs. Independent providers vs. Insurance company/self-insure options.
The following are some suggestions that were gleaned from the evaluation forms and discussion during the session.
1) Training Users…courses, workshops, …what works for whom? 2) Scientific Ethics…What are our responsibilities as lab Managers? 3) Mission statements/lab organization/lab business plans/future planning issues 4) Staffing issues including training, part-time student help, Training course T.A’s 5) Daily issues…prioritizing use, prioritizing service projects, dealing with difficult users/customers 6) Legal issues involving data ownership, confidentiality of data, handling of potential evidence. 7) Justification for new equipment 8) Commercial use of university facilities 9) Microscopy as a research tool…integration of industry or campus facilities in coordination with other core facilities. 10) Issues unique to Multi-user and/or Service facilities and how such facilities can coexist.
The first 2 received the greatest number of requests. However there may be more relevant topics now. Please rank the 3 of highest priority to you or suggest additional topics of interest. I will sift through all replies at the end of next week and narrow them down to 3. Also please feel free to supply the names of individuals to lead specific discussion topics. The format will be the same as the last session. A presenter will give a 10-15 min. introduction of the topic followed by ~30 min. open discussion.
Thanks, Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA18774 for dist-Microscopy; Thu, 18 Jan 2001 11:35:15 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA18769 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 18 Jan 2001 11:34:45 -0600 (CST) Received: from arcs4.cc.bbsrc.ac.uk (arcs4.cc.bbsrc.ac.uk [149.155.100.41]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id LAA18762 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 18 Jan 2001 11:34:34 -0600 (CST) Received: from arcs.cc.bbsrc.ac.uk by arcs4.cc.bbsrc.ac.uk with SMTP (PP) with ESMTP; Thu, 18 Jan 2001 17:32:12 +0000 Received: from exrout.cc.bbsrc.ac.uk by arcs.cc.bbsrc.ac.uk with SMTP (PP) with ESMTP; Thu, 18 Jan 2001 17:31:54 +0000 Received: from rs-exsrv1.res.bbsrc.ac.uk ([149.155.12.82]) by exrout.cc.bbsrc.ac.uk with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2232.9) id DFB0BSPQ; Thu, 18 Jan 2001 17:31:51 -0000 Received: by rs-exsrv1.res.bbsrc.ac.uk with Internet Mail Service (5.5.2232.9) id {DFCBTR77} ; Thu, 18 Jan 2001 17:31:50 -0000 Message-ID: {194E4E81F995D411989200508BDCD8F3212617-at-rs-exsrv1.res.bbsrc.ac.uk}
Hi Mike, Just a thought. When we tried to change to other metals we found that our sputterer would handle Au, Au/Pd but not Pt. Might be worth checking yours before spending serious money. ttfn, Chris
I am searching for a new TEM (hoping to write a grant to get funding for one anyway - since I am not independantly wealthy). The one we currently have is a JOEL 100S (and a crabby one, too boot).
I would like to know what is available. I would also like to know some approximate prices.
Please reply off-list!
Thank you. Susan J Rehorek, Ph.D. Department of Biology Slippery Rock University of Pennsylvania PA, 16057-1326
I was wondering if there are any stains/methods that would allow the identification of basal bodies or centromeres in a non-dividing cell. I would like to determine the basal body position in a cell without resorting to serial TEM. My thinking is that I might be able to do it in whole cells with the right marker. Any comments?
Majid, Try 50% sulfuric acid for 2 hours to overnight. I used this successfully some years ago on replicas from Xenopus oocytes. Frank
At 08:10 AM 1/18/01 -0600, Majid wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, EDS is very real and chip manufacturers go to great length (end expense) to avoid them. However, I think the mechanism is different:
EDS means, that something collects static electricity (by walking on a carpet, for example), then touches something that is at a very different potential. That's what happens if you touch a metal after walking on that carpet. The enormous potential difference create strong electric field that finally ionize the air and you get "zapped". The strong fields can destroy the electronics.
In an SEM the sample is usually grounded and does not see strong fields. True, the electrons are accelerated to 30 KV or more, but the sample does not see that, as the anode usually sits at ground level with the Cathode being at - 30 kV. What you need to be concerned about here is the kinetic energy that is transferred to the sample and can heat it up considerable. On the other hand, if the sample is not grounded, there will be fields developing. There are also other effects, such as residual organics getting "cracked" and forming a residue on the sample, but that's a different story.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Earl Weltmer [mailto:eweltmer-at-home.com] Sent: Thursday, January 18, 2001 7:24 AM To: ars-at-sem.com; 'Don Grimes'; microscopy-at-sparc5.microscopy.com
Hi All,
ESD damage on semiconductor samples is quite real & has been documented for years. When CMOS circuits first came out this was a real concern. Motorola & a number of other semiconductor companies has reams of papers on this subject.
Contact me offline & I can see if I can dig up the info.
Regards,
Earl
----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Don Grimes'" {microtoday-at-mindspring.com} ; {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 18, 2001 2:10 AM
Can anyone offer any tips for embedding rat tongue tissue in paraffin? I am having difficulty with complete infiltration (?) of the tissue. The tongue has a dense muscle core that becomes hard during the process causing shredding during sectioning. Any help would be much appreciated.
On a second note: This tissue has been injected with tritiated thymidine. Has anyone ever heard of loss of radioactive label during tissue post-fix or embedding?
thanks so much,
Susan Hendricks University of Virginia Department of Psychology PO Box 400400 Gilmer Hall Charlottesville, VA 22904
Thank you for the numerous responses to my query about which metals and operating conditions to consider for improving the grain size of the coatings we have been getting from our sputter coater. Below is a brief summary of the recommendations we have received. I will be experimenting with several of these items to see where we ultimately want to end up with this. I suspect that we will develop several protocols based on the requirements of the experiment. Again thank you.
1) Gold/Palladium definitely has a smaller grain and we should see a definite improvement by using it.
2) Platinum will have the finest grain size but it takes longer to deposit, potentially causing damage to sensitive samples.
3) Platinum/Palladium is sometimes used as an alternative to Chromium coating
4) Reduce the coating thickness as much as possible
5) Coat using higher vacuum / lower currents / lower voltage to reduce grain size
6) Pulse the deposition
7) Decrease sample surface temperature
8) Gold may be the best alternative for samples where resolution is not as much a factor as is sample integrity as the other metals require more energy / longer exposure to get an adequate coating. ====================================== Michael D. Standing BYU Microscopy Lab 401 WIDB Brigham Young University Provo, UT 84602
Interesting. What was it about the unit that precluded Pt? Did the maker supply Pt targets or did you just try your own? It would seem that if Pt target are factory supplied, they should work with the system. My Hummer does Au, Au/Pd and Pt very well. No problems.
gary g.
At 09:31 AM 1/18/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What I did many years ago, I treated the replica with enzymes to degradate the most of the sample and then I did clean the replica in 20-40% H2SO4. The enzymes you could use depends from nature of your contamination on the replica. I did use proteinase-K. It's very active enzyme if you need to have deal with proteins. I assume that, because your containerizations are survived even in chromic-mixture (difficult to impress to me) it may be some poly-sugars or something like that. So, you may try the particular enzymes. There are plenty of them in the Sigma Catalog.
You may also use full-strength chromic-mixture. In theory it may not affect the Pt-C replica. I don't like the bleach at all, personally. Plasma cleaning will affect the integrity of the carbon layer, I believe.
Good luck.
Sergey.
At 08:10 AM 1/18/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
I've never had a problem with freeze damage and I've done this a lot...I can't see anything obviously wrong in what you're doing..The few things I can think of are:
- wash your cell pellet in PBS with 0.12% glycine before putting them in the gelatin. This will quench free aldehyde groups from the fix and avoid crosslinking the gelatin (I'm not sure this would affect the sucrose infiltration..?? but..maybe..I never tried not quenching)
- avoid putting the cells in gelatin (if you can). If you have tissue there's certainly no need for gelatin. Cut the fixed tissue in to small cubes, wash in PBS/glycine for 15 minutes then infiltrate in 3 drops of 2.3 M sucrose at RT for a total of 15 minutes (30 minutes for tissue pieces), put a piece of the pellet/tissue on the pin and put into LN2.
( I only do gelatin embedding if I have cells that have been fixed in suspension for some reason - they will never stay together as a pellet in the sucrose! If you spin the cells down in a microfuge tube in the fixative and let them fix as a pellet, they will stay together really well without the gelatin (you can get the pellet out of the microfuge tube with a toothpick)
- If you need to do gelatin, try putting the gelatin embedded cells in 2.3M Sucrose (not step-wise) and incubate over night in the fridge.
good luck! don't hesitate to contact me if you have more problems.
Maria
At 10:26 AM 1/17/01 -0500, Leslie Cummins wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________
Maria Ericsson Harvard Medical School EM Facility 220 Longwood Avenue Boston, MA 02115 (617) 432 1698 maria_ericsson-at-hms.harvard.edu http://www.hms.harvard.edu/core/em.html
The kinetic energy referred to by Michael can be a very real problem for your sample in the SEM.
As far as the concern over Electrostatic Discharge damage goes, it may not look the same as that caused by somebody's finger discharging through the part, but it will be real. I have analyzed samples with both types of damage. In the SEM, floating nodes of circuitry may charge up from the beam. When they exceed the breakdown voltage of the dielectric, such as a CMOS gate oxide, a very large, very short in duration, current spike will occur and cause extreme localized heating. This causes a hole in the oxide, filled with alloyed silicon, causing a short circuit. Another way for this to happen, is for the interlevel dielectric to build up a charge of electrons, and then suddenly discharge through a part of the circuitry. Both of these mechanisms have given me problems while trying to localize resistive via contacts with Resistive Contrast Imaging in the SEM, where the specimen absorbed current is imaged. The defect "heals" when it is blown through.
Another problem with looking at circuits that you hope to operate after SEM observation, is the electron beam can "implant" electrons in the chips structures and affect the proper operation of the circuitry. This is especially true at the higher accelerating voltages.
I hope this helps.
Darrell Miles Failure Analyst IBM Microelectronics Analytical Services Fishkill, New York
} Yes, EDS is very real and chip manufacturers go to great length (end } expense) to avoid them. However, I think the mechanism is different: } } EDS means, that something collects static electricity (by walking on a } carpet, for example), then touches something that is at a very different } potential. That's what happens if you touch a metal after walking on that } carpet. The enormous potential difference create strong electric field that } finally ionize the air and you get "zapped". The strong fields can destroy } the electronics. } } In an SEM the sample is usually grounded and does not see strong fields. } True, the electrons are accelerated to 30 KV or more, but the sample does } not see that, as the anode usually sits at ground level with the Cathode } being at - 30 kV. What you need to be concerned about here is the kinetic } energy that is transferred to the sample and can heat it up considerable. On } the other hand, if the sample is not grounded, there will be fields } developing. There are also other effects, such as residual organics getting } "cracked" and forming a residue on the sample, but that's a different story. } } Michael } } Michael Bode, Ph.D.
Not really true as backscattered electrons as well as x-rays generated from the sample can have an energy upto the acceleration voltage. The anode is grounded to create the 30KeV field which accelerates the electrons. The electrons shoot through the hole in the anode, through lenses (and other things) all the way to the sample, at 30KeV. Check any source on how the SEM works or for that matter how a basic electron accelerator works.
As for the EDS (static discharge that is) question, why would a chip maker use SEM to analysis the effect of static discharge if the SEM could cause the effect in the first place. Check out the older chip makers data books, plenty of SEM photos of EDS effects. An SEM would be pretty useless for IC failure analysis if it caused EDS damage.
EDS is destructive from the current density, not the field density. Too much current through a sub-micron conductor blows up the conductor in the same way electrical wires vaporize under too much current. EDS protection is handled through shunting the current before it gets to the sensitive areas. Most modern ICs are EDS protected, some (like IO drivers) more than others. Old hat to chip makers and not much expense compared to the rest of the design.
I would suggest this, try it. If you have EDS problems you will see the results. If you see the results, send the design back to R&D for further development. If you are nervious about 30KeV, use a lower voltage, that's what it is there for.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
} } } Do you know or can you ask your readers about the possibility of } } } Electrostatic Discharge damage caused by an SEM? Is this a problem? My } } } company is reluctant to have me examine flight-ready devices in the SEM } } } because of this possibility. I can't find any information on the } } } subject. Thanks for any help you can give me.
Virginia , if you are suggesting to examine these devices and then install. Very bad idea for any sort of testing besides operational on shipping product. This is never done for production that goes out the door. That's what product testing is all about. You take samples from production, then subject them to things that they will see over their life. If they fail, a statistical analysis will tell you their failure rate. Or if they cannot fail (because of were they are used), tells you to try again.
Flight-ready devices have very specific rules and regulations, I can understand their reluctance. Keeps things in the air, in the air. Why do you need to examine by SEM? Do you need to examine every one or will a statistical sample of production devices work as well. IC makers don't EDS test every chip they ship yet they are EDS rated. They test samples from pre-production and then test production samples every now and then. The tested devices are not returned to production.
If these devices are expensive (most flight-ready devices are) and your need for examination is important, then the company should have no problem pulling some production devices for you to use. Once used, they do not ship but are stored or destroyed. If you company says it's too expensive, then you have a managment problem because product testing costs should have been built into the selling price of the devices. It's part of NRE, non-returnable expenses.
For a flight-ready device example, say you have a box of electronic that must survive 100Gs. You don't subject every box to 100Gs and say ship the ones that survived. You design for 100Gs. You test for 100Gs. You start production and pull samples for testing at 100Gs. Over life of the production and whenever the production process changes, you test for 100Gs. Everytime you test, survive or fail, you open the box to check if there is an unseen problem that will cause a future failure because you performed the test. Product testing 101.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
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Usually 5% of the lot is pulled for testing. The evaluation for the entire lot is based upon the test results of the representative 5%.
Sometimes examining a part under the SEM will cause damage depending upon the construction of the part. The damage is artifact and has nothing to do with the part integrity. I think it is this damage that Raytheon is most concerned.
The emphasis for SEM examination of semiconductor has shifted from 15KV to 20KV, to lower (up to 2KV) accelerating voltages. Resolution at low KV suffers and hence the untilization of field emission guns.
Regards All,
Earl
----- Original Message ----- } From: "Scott D. Davilla" {davilla-at-4pi.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 18, 2001 4:30 PM
Majid:
The common cleaning methods for FF replicas include: Conc. Bleach, 70% Sulfuric acid, or 50%Chromic acid. These work for almost everything I have encountered. For the others I recommend Fisher Scientific Glass Cleaning Solution*. Start with 10% increasing to 50% overnight in a humidified chamber with a cover.
If you don't take care to slowly increase the conc. of ANY of the cleaning solutions you take the chance of "cooking" the remaining biological material to the replica. This maybe what is happening. Several labs I work with are using Xenopis oocytes and don't encounter problems with the replicas and they use only Bleach. Plasma cleaning replicas may also "cook" the replica too but I don't know for certain.
I think the brittleness you are encountering is related strictly to the replication process and not the cleaning. I would need more information on your protocol to comment on what might be causing it. Pt/C replicas are stabile in all of the solutions mentioned above for extended cleaning times.
* I have no financial interest in Fisher Chemicals or Products.
Al Coritz Sales & Service Manager RMC-Boeckeler Instruments 4650 S. Butterfield Dr. Tucson, AZ 85714 Voice: 520-745-0001 Cell: 520-465-3598 Fax: 520-745-0004 Email:Al-at-Boeckeler.com Website:RMCProducts.com
-----Original Message----- } From: Majid [mailto:majid.ghoddusi-at-anu.edu.au] Sent: Thursday, January 18, 2001 7:11 AM To: Microscopy-at-sparc5.microscopy.com
Hello all
We are preparing Pt-C and C replicas from different biological material. For cleaning the obtained replica we either use bleach (5.2%) or chromic acid (10, 20 followed by 40%) with washes in between. The protocol usually works well for most stuff but when it comes to oocytes from Xenopus we are usually left with a lot of organic material still attached to the replica. If we leave the replica in cleaning agent for a long time we end up with a very brittle replica which can easily break.
Oocytes have got a reputation for being very sticky and difficult to clean. We are considering to use plasma cleaning instead but I don't know what to expect from that. I would appreciate it if you could share your 'tips and tricks' with us.
Majid Ghoddusi
............................................................... Majid Ghoddusi, PhD Protein Dynamics Unit Department of Chemistry Australian National University ACTON 0200 Canberra Tel: (02) 6125 4190 Fax: (02) 6125 0760 e.mail: majid.ghoddusi-at-anu.edu.au web site: http://langevin.anu.edu.au/ .......................................................................... .........
Hi Karen - I've worked with ciliates (i.e., eukaryotic microorganisms -- one common example is Paramecium), and there are several silver stains that, when properly executed, allow the user to clearly visualize basal bodies under LM brightfield. They are: Klein silver stain (using silver nitrate); Chatton-Lwoff silver stain (using a salinated gel in combo with silver nitrate); Rio Hortega ammoniacal silver stain (using a combo of ammonium hydroxide and silver nitrate), and Protargol stain (using proteinaceous silver nitrate). There are protocols for these stains in a protozoology book by Kirby (I forget the title offhand). Any of these stains may help you. Good luck with the basal body visualization! Nelson Conti
On Thu, 18 Jan 2001 12:47:08 -0600, Karen.Pawlowski-at-worldnet.att.net wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi all, | | I was wondering if there are any stains/methods that would allow the | identification of basal bodies or centromeres in a non-dividing cell. | I would like to determine the basal body position in a cell without | resorting to serial TEM. My thinking is that I might be able to do | it in whole cells with the right marker. Any comments? | | Karen Pawlowski |
164 Ferne Court Palo Alto, CA 94306 (650) 494-0472 [NelsonC51-at-excite.com]
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one way that you could improve the cooling properties of your liquid nitrogen is by 'slushing' it. This simply involves supercooling liquid nitrogen by using latent heat of evaporation. A simple rotary pump and vacuum chamber is used to evaporate gas off the liquid until a slush of solid nitrogen is produced. If air is leaked into the chamber the solid nitrogen melts and the process should be repeated 2 or 3 times to thoroughly cool the nitrogen. The resulting liquid/slush should be handled with great care because it does not boil when warm objects are plunged into it (i.e. it will freeze specimens and you more quickly). It can be used for a few minutes after production and can be tested simply by dropping small metal objects in (if they 'fizz' it's no longer cool enough).
If you intend to try this you will need to experiment with quantities and containers (about 100-200ml in a 200-400ml PTFE or metal beaker) and take care that the nitrogen doesn't all spurt out when the solid forms on its surface. You will need a vacuum chamber design where chilling and nitrogen spillage will not cause damage, you can view the process through a window and bleed air in quickly. I have used freeze drying units and a home/lab-made chamber made out of an old film desiccator and they both worked well. If you're careful and have some form of insulated drip tray you might try an ordinary vacuum coater with just the roughing/rotary pump.
I'm sure there is plenty of published information on this technique (perhaps in a Hayat book or other cryo technique texts) and I apologise if you were already aware of it. If my memory serves me well, the last paper I looked at stated that pentane etc. were better cryogens but that slush was a lot better than liquid nitrogen.
Good luck
Malcolm Haswell e.m. unit University of Sunderland UK
Leona Cohen-Gould wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Leslie, } One thing I can think of, is that you shouldn't drop your samples } into the liq. Nit. LN2 is a poor cryogen in that it boils when warm } things are placed into it, creating a pocket of nitrogen gas around } the sample, that acts as an insulating layer. Most of us do use } nitrogen to freeze our samples. I always hold the pin end of the } stub in a hemostat-type clamp and wiggle it as it freezes to try to } ensure an exchange of the cryogen and avoid too much of a gas bubble } around the sample. I'm sure you will get many other suggestions, } since there are so many of us out there doing this with varying } degrees of success. } Good luck, } Lee } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175
} } } } The orignal question was } } } } } } } Do you know or can you ask your readers about the possibility of } } } } } Electrostatic Discharge damage caused by an SEM? Is this a problem? } My } } } } } company is reluctant to have me examine flight-ready devices in the } SEM } } } } } because of this possibility. I can't find any information on the } } } } } subject. Thanks for any help you can give me. } }
Having worked on aircraft parts you will never get to test a flight ready part and get it declared flight ready again. I expect an inspector will destroy it in some way when you are finished with it. If you think about it a little you wouldn't want it any other way. The device was not designed to be run through a SEM so even if it was totaly safe it would be out of the queston.
If you need to look at one look at one that has been used in other testing or has failed inspection and can not be reworked. These happen on occasion just have them hold one for you.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
sorry that I answer your question a bit late! Some weeks ago you asked about recipes for LR White embedding of plant tissue.
Well, here are some references:
Lonsdale J.E., McDonald K.L., Jones R.L. (1999): High pressure freezing and freeze substitution reveal new aspects of fine structure and maintain protein antigenicity in barley aleurone cells. Plant Journal 17(2): 221-229.
Takahashi, H., Kuroiwa, H., Miyagishima, S., Toda K., Itoh R., Kuroiwa T. (1997): Improved procedure for immunogold electron microscopy: Rapid-freeze substitution with absolute acetone. Cytologia (Tokyo) 62(3): 303 ? 308
This one is with LR Gold: Staff, I.A., Schappi, G., Taylor P.E. (1999): Localisation of allergens in ryegrass pollen and in airborne micronic particles. Protoplasma 208(1-4): 47 ? 57.
Hope that helps,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Hello } My problem is: after revealing the semithin slices (10 um), I can see } eye-naked the blue staining, but then it fades away in 10 to 30 } minutes. Can it be possible that the glycerol-PBS that I use as } mounting media has something to do, or it is maybe the PBS in which I } rinse the slides after revealing, to clean the excess and toxic DAB ? } } We suspect the PBS may have some contaminating elements which may } reduce the DAB so it re-dissolves, or the glycerol is too old (it was } bought in 1997) and has turned acid so it gives protons to the media so } DAB re-dissolves. } Has it ever happened to anyone? Thank you. } } Albert Cardona. } University of Barcelona. Spain.
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Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Email: ldb2-at-ix.netcom.com Name: Larry Bell School: Westfields High School State: VA
Question: My son built a polarized light microscope and has been trying to observe the liquid crystal properies of DNA for his 10th grade science fair project. We've been all over looking for books and materials on the subject and haven't had much luck. We've seen great pictures of the 3 liquid crystal phases exhibited by DNA, but can't find information on preparing the sample and the slides. His science teacher gave him a simple procedure to extract DNA from peas (or bananas) using a blender, meat tenderizer and alcohol. The DNA rises to the top after the alcohol is added. He tried allowing a drop of DNA/Alcohol mixture evaporate under the crossed polarizer, but no changes have been observed. I'm told he needs to "dissolve the DNA in a buffered saline solution prior to evaporation". Can you advise him on a simple process to prepare the sample, and offer advice on how to obtain or make the buffered saline solution?
Can you help us with a simple process to prepare a DNA sample for analysis by evaporation?
I have used all of the aforementioned replica cleaning reagents. One that hasn't been mentioned yet, and that is very good for cleaning Pt/C replicas is HF. The method is like all the rest, except that the HF has to be in plastic containers (small petri dish, usually). Cleaning time can be up to overnight, wash by transferring across several water washes, and pick up on the grid. This is the method of choice for high resolution shadowed/replicas protein preps. I have used it on other specimens as well, and think that if necessary you could try it on yours. Hope this helps.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Thu, 18 Jan 2001 21:57:52 -0700, Al Coritz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Majid: } } The common cleaning methods for FF replicas include: Conc. Bleach, 70% } Sulfuric acid, or 50%Chromic acid. These work for almost everything I have } encountered. For the others I recommend Fisher Scientific Glass Cleaning } Solution*. Start with 10% increasing to 50% overnight in a humidified } chamber with a cover. } } If you don't take care to slowly increase the conc. of ANY of the cleaning } solutions you take the chance of "cooking" the remaining biological material } to the replica. This maybe what is happening. Several labs I work with are } using Xenopis oocytes and don't encounter problems with the replicas and } they use only Bleach. Plasma cleaning replicas may also "cook" the replica } too but I don't know for certain. } } I think the brittleness you are encountering is related strictly to the } replication process and not the cleaning. I would need more information on } your protocol to comment on what might be causing it. Pt/C replicas are } stabile in all of the solutions mentioned above for extended cleaning times. } } * I have no financial interest in Fisher Chemicals or Products. } } } Al Coritz } Sales & Service Manager } RMC-Boeckeler Instruments } 4650 S. Butterfield Dr. } Tucson, AZ 85714 } Voice: 520-745-0001 } Cell: 520-465-3598 } Fax: 520-745-0004 } Email:Al-at-Boeckeler.com } Website:RMCProducts.com } } } } -----Original Message----- } } From: Majid [mailto:majid.ghoddusi-at-anu.edu.au] } Sent: Thursday, January 18, 2001 7:11 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Freeze-Fracture replica cleaning } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hello all } } We are preparing Pt-C and C replicas from different biological material. For } cleaning the obtained replica we either use bleach (5.2%) or chromic acid } (10, 20 followed by 40%) with washes in between. The protocol usually works } well for most stuff but when it comes to oocytes from Xenopus we are usually } left with a lot of organic material still attached to the replica. If we } leave the replica in cleaning agent for a long time we end up with a very } brittle replica which can easily break. } } Oocytes have got a reputation for being very sticky and difficult to clean. } We are considering to use plasma cleaning instead but I don't know what to } expect from that. I would appreciate it if you could share your 'tips and } tricks' with us. } } Majid Ghoddusi } } } } ............................................................... } Majid Ghoddusi, PhD } Protein Dynamics Unit } Department of Chemistry } Australian National University } ACTON 0200 } Canberra } Tel: (02) 6125 4190 } Fax: (02) 6125 0760 } e.mail: majid.ghoddusi-at-anu.edu.au } web site: http://langevin.anu.edu.au/ } ......................................................................... } ......... } } } }
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First, a question: are you embedding the whole tongue or a cross section (or cross sections)? We routinely process rat tongue as part of the tissue workup on whole animal collections. We collect a cross section approximately 7mm long and embed to view the cross section. Processing is done with the same schedule that we use for all the rest of our rat tissue. Dehydration/clearing are at 45 min steps except for the last 100% alcohol and clearant (those are 60 min each). We use either two 1 hour 30 min steps in paraffin (Shandon Hypercenter XP) or two 45 min and two 60 min steps (VIP E300). The only differences I can see between the two processors is that the skins and cns tissues tend to be less well embedded through the Shandon two-stage paraffin vs the VIP four-stage paraffin. Tongue embeds equally well in either (as does other dense tissues)--the problem with skin (especially from female rats or very old animals--either sex) and cns is the high lipid/fat content and the ability to remove/replace that more effectively with the additional paraffin infiltration steps. The other thing I do is to make sure that my paraffins are not contaminated with clearant (even the first stage). If the paraffin feels "slippery" when rubbed between finger and thumb, dump it and replace with fresh paraffin. Hope this helps.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Thu, 18 Jan 2001 16:15:54 -0500, Sue Hendricks wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone offer any tips for embedding rat tongue tissue in paraffin? I } am having difficulty with complete infiltration (?) of the tissue. The } tongue has a dense muscle core that becomes hard during the process causing } shredding during sectioning. Any help would be much appreciated. } } On a second note: This tissue has been injected with tritiated thymidine. } Has anyone ever heard of loss of radioactive label during tissue post-fix or } embedding? } } thanks so much, } } Susan Hendricks } University of Virginia } Department of Psychology } PO Box 400400 } Gilmer Hall } Charlottesville, VA 22904 } } 804.982.4755 } 802.982.4785 [fax] } sjh2e-at-virginia.edu } }
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Thanks, Allen, for pointing out some of the issues here. By the way, It's "Wehnelt", not "Wienault".
You are probably right about the ESD vs. EDS name. However, it was clear to me what the original question was about.
Someone pointed out, that the cause of ESD (!) failure in electronic devices is, that the fields increase to beyond the breakdown strength of the devices, which causes a high and sudden current to flow through the parts where the fields are highest, which in turn heats and destroys the sample. I think, that is right. It's the fields that reach critical strength, which then causes the current flow and destruction of the device. That's what I meant when I said you get "zapped".
Now, about the electron microscope. Obviously I don't want to pronounce, that electron microscopy is impossible. That would be stupid, wouldn't it? However, as you said yourself, the sample is at ground potential, or very near it, and so is the anode. In other words, there is not electric potential (or very little) between the anode and the sample and thus no electric field. Since the sample is grounded, as is the rest of the microscope, there is no electric field between those, either. The energy of an electron in an electric field is proportional to the electric field (actually proportional to the square of the field, if my memory serves me). No field - no electric energy. Of course that does not mean that the electrons don't have energy. They have quite a lot of energy, on the order of 20keV, by "falling" through the electric field between cathode and anode and not being stopped by the anode. Since the sample is grounded (or should be), the electric effects of the electron beam on the grounded sample will be cancelled. What you have to contend with is the kinetic energy (20 keV/electron), which is transferred to the sample.
Now, I have always talked about grounded samples. That does not mean that electric fields cannot be generated by the electron beam within the sample. Obviously, you put electrons in the sample and they have to move out of the sample. If there is any kind of resistance (an oxide layer, etc.), that will result in charging and an electric potential.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Allen R. Sampson [mailto:ars-at-sem.com] Sent: Friday, January 19, 2001 2:44 AM To: 'Mike Bode'
Hi, Many thanks for the suggestions. We have an Oxford Instruments 1500 attached to an SEM. Oxford themselves say that this model will only sputter Au or Au/Pd, although they supply all types of target. It can be upgraded to a 1500HF at £11,000 plus tax which will do Pt but they say we would also need to swap the rotary vacuum pump for a turbo. We compromised and bought an Au/Pd target. We find cracking open the valve to the SEM chamber to get a sniff of the higher vacuum helps deposition. ttfn, Chris
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Microscopists, We have some students doing projects where GAS staining is used as a genetic marker in arabadopsis roots. These roots are very small and must then be fixed embedded and thick-sectioned so that the distribution of the blue GUS staining can be observed with the light microscope.
The staining tends to fade badly with most embedding procedures. We have found that if samples are only run up to 95% ETOH and then embedded, the staining intensity is preserved. Thus only resins which tolerate 5% water can be used. We have successfully used technovit but this resin is very difficult to cut. Has anyone had experience with this stain and other resins that might preserve the stain intensity but is easier to handle (and hopefully not as expensive!). Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id PAA23212 for dist-Microscopy; Fri, 19 Jan 2001 15:54:41 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id PAA23206 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 19 Jan 2001 15:54:10 -0600 (CST) Received: from aurora.uaf.edu (aurora.uaf.edu [137.229.18.100]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id PAA23198 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 19 Jan 2001 15:53:59 -0600 (CST) Received: from emlab.alaska.edu (kim.geology.uaf.edu [137.229.53.96]) by aurora.uaf.edu (8.9.3/8.9.3) with ESMTP id MAA28312 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 19 Jan 2001 12:52:27 -0900 (AKST) Message-Id: {4.3.2.7.2.20010119123432.00ad6180-at-aurora.alaska.edu} X-Sender: fnksd1-at-aurora.alaska.edu X-Mailer: QUALCOMM Windows Eudora Version 4.3.2
Fellow Listers, Our Zeiss 109 recently developed a large vacuum leak in the fiber optic panel (Between the column and the camera). I am told a new one runs approximately $6000.00. We are no longer on service contract, but my old service technician at LEO may already have a line on a used one. Above and beyond the call of duty, if you ask me. In case that falls through I thought I'd see If anyone out there might know of a likely source. Thanks in advance You are all so amazingly resourceful
Kim DeRuyter Histology and Electron Microscopy Technician 900 Yukon Dr. Fairbanks, Alaska 99775-0180 907-474-5452
Hi Kim, I think there is a 35mm camera port on that microscope? You might be able to blank off the bottom plate and pick up a second hand 35mm camera. Good luck Sally
Dr Sally Stowe Facility Coordinator, ANU EMU Box 475 Canberra ACT 2601 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 or 6125 8525 http://www.anu.edu.au/EMU
} } } Kim DeRuyter {fnksd1-at-uaf.edu} 01/20/01 08:51am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Fellow Listers, Our Zeiss 109 recently developed a large vacuum leak in the fiber optic panel (Between the column and the camera). I am told a new one runs approximately $6000.00. We are no longer on service contract, but my old service technician at LEO may already have a line on a used one. Above and
beyond the call of duty, if you ask me. In case that falls through I thought I'd see If anyone out there might know of a likely source. Thanks in advance You are all so amazingly resourceful
Kim DeRuyter Histology and Electron Microscopy Technician 900 Yukon Dr. Fairbanks, Alaska 99775-0180 907-474-5452
I need to obtain some certified standards for carbon in steel matrix to do quantitative work using WDX. The problem is finding material that is homogeneous on a micro-scale. I am trying to cover a range of 0.1 to 2.0 Wt. % carbon. No luck thus far with NIST or Brammer.
Does anyone know of a supplier who can produce and sell such material?
Thanks in advance.
Jim Koncki Senior Project Chemist Torrington Co./Ingersoll-Rand
Does anyone use an Alden model 9315 CTP direct thermal printer to print images? We have obtained this printer without drivers and the company appears to be out of business. Thanks, Lynne C. Garone Polaroid Corp. W4-1D 1265 Main St. Waltham, MA 02451-1714 (781) 386 -1446 (781) 386 -0378 GaroneL-at-Polaroid.com
} From: "Kim DeRuyter" {fnksd1-at-uaf.edu} } Fellow Listers, } Our Zeiss 109 recently developed a large vacuum leak in the fiber optic } panel (Between the column and the camera). I am told a new one runs } approximately $6000.00. We are no longer on service contract, but my old } service technician at LEO may already have a line on a used one. Above and } beyond the call of duty, if you ask me. In case that falls through I } thought I'd see If anyone out there might know of a likely source. } Thanks in advance } You are all so amazingly resourceful } A free wanted add on www.labx.com turns up a lot of hard to find stuff and the price it right.
I have no connection with labx except using their service to find some really hard to find stuff.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
I think that the emphasis is not quiet right. If you have resolution problems due to the coating in the common SEM magnification range at say up to x60k, chances are, its the thickness of the coating and not the grain-size. The analogy with a blanket of snow should be clear to all of you in frigid climates: A dusting of snow contours every pebble, whereas a 300mm layer would hide all small features. You cannot expect to see much detail using a thick coating. A thin coating may not be continuos because of grain size and a thin coating may lead to charging effects and these have to be addressed by various means. Finer grain size gives a more continuous coating, but if the coating is thick the grain-size scarcely matters in terms of resolution. The difference in grain size between Au and Au/Pd is marginal and was first given as a theoretical advantage when SEM specimens were coated by evaporation. Sputtering gives lower grain sizes than evaporation for any given metal, but I guess that the relative grain-size remains. I rather doubt that you will find any real advantage after switching from Au to Au/Pd targets. Pt would give an advantage at high SEM mags, but even then, part of the effect may be the thinner coating that Pt gives when compared with Au. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, January 19, 2001 7:29 AM, Michael D. Standing [SMTP:Michael_Standing-at-byu.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers: } } Thank you for the numerous responses to my query about which metals and } operating conditions to consider for improving the grain size of the } coatings we have been getting from our sputter coater. Below is a brief } summary of the recommendations we have received. I will be experimenting } with several of these items to see where we ultimately want to end up with } this. I suspect that we will develop several protocols based on the } requirements of the experiment. Again thank you. } } 1) Gold/Palladium definitely has a smaller grain and we should see a } definite improvement by using it. } } 2) Platinum will have the finest grain size but it takes longer to deposit, } potentially causing damage to sensitive samples. } } 3) Platinum/Palladium is sometimes used as an alternative to Chromium } coating } } 4) Reduce the coating thickness as much as possible } } 5) Coat using higher vacuum / lower currents / lower voltage to reduce grain } size } } 6) Pulse the deposition } } 7) Decrease sample surface temperature } } 8) Gold may be the best alternative for samples where resolution is not as } much a factor as is sample integrity as the other metals require more energy } / longer exposure to get an adequate coating. } ====================================== } Michael D. Standing } BYU Microscopy Lab } 401 WIDB } Brigham Young University } Provo, UT 84602 } } e-mail: Michael_Standing-at-byu.edu } phone: (801) 378-4011 } fax: (801) 378-3937 } ====================================== } }
Thank you Grace It was my first question here and it amazes me how fast answers came from everywhere. Yes, I'm using niquel amonium sulfate as DAB enhancer, and then I mount the slides with glycerol-PBS 50%, since the slides are 8-well ones -pretty expensive- and we re-use them hundreds of times. It would not be possible to do so with permanent mounting media as DPX or permount. Anyway, there is no point in keeping the screening results ... these immunostainings are used screen monoclonal antibodies produced by myself. But, the issue is, it has always been possible to mount the slides in glycerol-PBS with no loss-of-stain problems -at least, the blue stain lasted 2 days. But now we are losing the staining in 30 minutes, even less, sometimes even 10 minutes. That is why I was asking if the glycerol aged more acid had something to do, or the PBS. Thank you. Oh, by the way, I accidentaly deleted the last 10 mails, one of them also including an answer to my request. If anybody could be so kind as to resend it to me to planaria2000-at-yahoo.com I would really appreciate it.
Albert Cardona University of Barcelona. Spain.
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Thank you Grace It was my first question here and it amazes me how fast answers came from everywhere. Yes, I'm using niquel amonium sulfate as DAB enhancer, and then I mount the slides with glycerol-PBS 50%, since the slides are 8-well ones -pretty expensive- and we re-use them hundreds of times. It would not be possible to do so with permanent mounting media as DPX or permount. Anyway, there is no point in keeping the screening results ... these immunostainings are used screen monoclonal antibodies produced by myself. But, the issue is, it has always been possible to mount the slides in glycerol-PBS with no loss-of-stain problems -at least, the blue stain lasted 2 days. But now we are losing the staining in 30 minutes, even less, sometimes even 10 minutes. That is why I was asking if the glycerol aged more acid had something to do, or the PBS. Thank you. Oh, by the way, I accidentaly deleted the last 10 mails, one of them also including an answer to my request. If anybody could be so kind as to resend it to me to planaria2000-at-yahoo.com I would really appreciate it.
Albert Cardona University of Barcelona. Spain.
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For your information, thought you all might be interested to know what is happening in microscopy in this sector. The sender is the head of plant / food em microscopy at AG Canada in Dartmouth NS I think.
Tim
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Food Structure & Functionality Symposium 2001 An international symposium leading food structure & Functionality studies through the 21st century *webaddress at the AOCS site http://www.aocs.org/member/division/fsff/index.htm*
Being held in conjunction with the , May 13-16, 2001, Minneapolis Convention Center, Minneapolis, Minnesota, USA. For information, e-mail us at: meetings-at-aocs.org
The symposium has two themes: * New and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * Food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Program (confirmed as of January 12th, 2001)
May 13th - Short Courses (short courses will run for a full day, and will run concurrently)
1) Food Contaminants. Contact person - Mark Auty 2) Specific Labeling in Foods. Contact person - Marcel Paques
May 14th-16th inclusive - Technical sessions 6 sessions will run over 3 days
Morning Symposium opening Plenary lecture - Food Quality andwhy the Structure matters. P. Lillford, Unilever Research, Colworth House UK
Session 1: Dairy Products and Fat Based Foods Session - chairs M. Auty and M. Paques
Milk protein polysaccharide interactions in aqueous solutions and oil-in-water emulsions. H. Singh*, Y. Hemar & P. Munro, Institute of Food, Nutrition and Human Health Massey University, Palmerston North, New Zealand
Structural functions of dairy ingredients in products formulated with taro flour. C. Onwulata, USDA/ARS,Eastern Regional Research Center,Wyndmoor, PA 19038
Confocal imaging of galactomannan mode of action in recrystallisation of ice in model ice cream D. Ferdinando, Unilever Research Colworth House, UK
Heating of Food Proteins in a Closed System at High Temperature N. Kitabatake, Kyoto University, Japan
Milk Protein - molecular components and functional properties. N. C. Ganguli, Indian Dairy Association
Afternoon Session 2: Food Safety - chairs J. W. Arnold and R. Droleskey Prevention of Foodborne Illness Through Sanitation and Processing Technology J.W. Arnold; USDA/ARS, Russell Research Center; Athens, GA 30604
PreHarvest Intervention Strategies to Control Foodborne Pathogens in Poultry J. A. Byrd; USDA/ARS/SPARC; College Station, TX 77845
Interactions of Competitive Exclusion Cultures with the Intestinal Mucosa of Newly Hatched Chicks R. Droleskey; USDA/ARS/SPARC; College Station, TX 77845
Adhesion of Salmonella on Alfalfa Sprouts A. Chartowski; USDA/ARS, Western Regional Research Center; Albany, CA 94710
Growth of Fusarium moniliforme Dependent upon Corn Tissue Type I. E. Yates; USDA/ARS, Russell Research Center; Athens, GA 30604
Dedicated poster viewing 4:00-6:00PM
Evening: Round Table Discussion - topic to be announced
Tuesday, May 15th, 2001 Morning Session 3: New Methods and Techniques for Food Structure and Functionality Analysis -chair K.Groves
Diffusing wave spectroscopy - a new and non-invasive method for the investigation of the structure, dynamics and interactions in complex food systems M. Alexander* and P. Schurtenberger
Food: How complex can it be? E. Esselink ,Unilever Research Vlaardingen, The Netherlands
Immunolocalization of Transgenic Protein in Wheat Endosperm M. L. Parker *, E. Stoger, R. Casey and P. Christou, Institute of Food Research, Norwich, England.
Structure/Function relationships through microrheology. M. Paques, Wageningen Centre for Food Sciences/Unilever Research Vlaardingen
Specific labeling techniques for foods J. Leunissen, Aurion: Immuno-Gold Reagents & Accessories, The Netherlands
Food Structure and Functionality Division Luncheon. Dr. Brian Brooker (Institute of Food Research, England * retired) will be presented with the Food Structure and Functionality Division award and will give a presentation entitled: Fat crystals - the importance of being small.
Afternoon Session 4: Agricultural Applications of Microscopy and Imaging Session- chairs D.F.Wood and P. Allan-Wojtas
The Utility of Sorting in Agriculture. H. J. Arnott, Department of Biology, University of Texas at Arlington, Texas
The Potential for Automatic X-ray Sorting of Insect Infested Grain R. Haff . USDA - ARS - WRRC, Albany, California.
Automated Sorting of Almonds with Embedded Shell by Laser Transmittance Imaging T. Pearson* R. Young, USDA - ARS - WRRC, Albany, California.
Use of a GFP-transformed strain of Fusarium graminearum to study ear rot development in corn S. S. Miller. , AAFC - ECORC, Ottawa, Ontario, Canada.
Popping modifies endosperm structure and improves digestibility in maize and sorghum. M.L. Parker, Institute of Food Research, Norwich, England.
Microstructural Changes in Rice During Cooking D. Wood* and P.C. Yu . USDA - ARS - WRRC, Albany, California.
Evening: Food Structure and Functionality Division Member meeting
Wednesday, May 16th, 2001 Morning Session 5: Ingredients and Food Processing - chairs D. Kittleson and J. Charbonneau
Water continuous fat crystal networks in ice cream induced by unsaturated monoglycerides N. M. Barfod , Danisco Cultor, Brabrand, Denmark
High pressure application fo food systems and its impact on functional ingredients B. Tauscher*, P. Butz,and A.F. Garcia, Federal Research Center for Nutrition, Karlsruhe, Germany
Specificity and application of lipolytic enzymes in bread making processes T.B. Frandsen, T. Spendler, G. Budolfsen, L. Christiansen and J. B. Neilsen, Novozymes A/S, Bagsvaerd, Denmark
Afternoon Session 6: Colloidal and Interfacial Sciences - chairs D. Pechak and M. Paques
Rheology and Structure of Particulate Protein Gels T. van Vliet, Wageningen Centre for Food Sciences,Wageningen, The Netherlands
Posters:
1. In situ study of the effect of heating on dough components using Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR)
O. Sevenou, S.E. Hill, I.A. Farhat and J.R. Mitchell. Division of Food Sciences, University of Nottingham, Loughborough, UK
2. Antioxidative activity of the crude extract of lignan glycosides from sesame meal
Y-S Shue and L.S. Hwang, Graduate Institute of Food Science and Technology, National Taiwan University, Taiwan, R.O.C.
3. Near Field Microscopy of phase separation in a mixed interfacial protein/surfactant film
A. P. Gunning, A.R. Mackie, A.R. Kirby and V.J. Morris, Institute of Food Research, Norwich Research Park, Norwich, UK
Contact information for the chairs is shown below, in alphabetical order:
Paula Allan-Wojtas Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre, Kentville, Nova Scotia, Canada B4N 1J5 Tel: (902) 679-5566 FAX: (902) 679-2311 email: allanwojtasp-at-em.agr.ca
Judy Arnold USDA -ARS - RRC 950 College Station Rd. P.O. Box 5677 Athens, GA 30604-5677 USA Tel: (706) 546-3515 FAX: (706) 546-3068 email: jarnold-at-ars.usda.gov
Mark Auty Dairy Products Research Centre TEAGASC Moorepark, Femoy, Co. Cork Ireland Tel: 011-353-25-42447 FAX: 011-353-25-42340 email: mauty-at-moorepark.teagasc.ie
James E. Charbonneau National Food Processors Association Food Chemistry and Packaging Department 1401 New York Ave, NW Washington, D.C. 20005 USA Tel: (202) 639-5972 FAX: (202) 639-5991 email: jcharbo-at-nfpa-food.org
Kathy Groves Leatherhead Food Research Association Randalls Road, Leatherhead Surrey KT22 7RY England Tel: 44 0132 822330 FAX: 44 0132 386228 email: kgroves-at-lfra.co.uk
Tony McKenna New Zealand Dairy Institute Private Bag 11 029 Palmerston North, New Zealand Tel: 011 64 6 350 4649 FAX: 011 64 6 356 1476 email: tony.mckenna-at-nzdri.org.nz
Marcel Paques Wageningen Centre for Food Sciences/Unilever Research Vlaardingen P.O. Box 20, 6710 BA Ede The Netherlands Tel: 011 31 318 659690 FAX: 011-31-318 650400 email: paques-at-nizo.nl
David Pechak Kraft Technology Centre 801 Waukegan Road Glenview, IL 60025 USA Tel: (847) 646-4808 FAX: (847) 646-3864 email: dpechak-at-kraft.com
Delilah Wood USDA - ARS - WWRC 800 Buchanan Street Albany, CA 94710 USA Tel: (510) 559-5653 FAX: (510) 559-5777 email: wood-at-pw.usda.gov
You might try Geller Microanalytical Lab in Peabody, Mass. The last phone number I had for him was 508-535-5595. If he doesn't have it, he would know where to direct you.
-----Original Message----- } From: "Jim_Koncki-at-Ingersoll-Rand.com"-at-sparc5.microscopy.com [mailto:"Jim_Koncki-at-Ingersoll-Rand.com"-at-sparc5.microscopy.com] Sent: Friday, January 19, 2001 5:14 PM To: Microscopy-at-sparc5.microscopy.com
Hi All:
I need to obtain some certified standards for carbon in steel matrix to do quantitative work using WDX. The problem is finding material that is homogeneous on a micro-scale. I am trying to cover a range of 0.1 to 2.0 Wt. % carbon. No luck thus far with NIST or Brammer.
Does anyone know of a supplier who can produce and sell such material?
Thanks in advance.
Jim Koncki Senior Project Chemist Torrington Co./Ingersoll-Rand
there is (probably not a "routine method") a really good paper on embedding Cornus wood in Spurr:
Ristic, Z., Ashworth E.N. (1995): Response of xylem ray parenchyma cells of supercooling wood tissues to freezing stress: Microscopic study. International Journal of Plant Sciences. 156(6): 784-792.
The preparation there was done by plunge freezing followed by Freeze Substitution.
Hope that helps,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
A friend wants to examine Mg-based alloys in TEM and warned us that in a previous lab this alloy was expelled from the microscope because it caused contamination by sublimation. I have no experience with these materials. Can anyone give me more detailed info if this is a real danger?
Thanks in advance:
János
Dr. habil, Janos L. Labar Research Institute for Technical Physics and Materials Science H-1121 Budapest, Konkoly-Thege u. 29-33 Postal address: H-1525 Budapest-114, Po Box 49 Tel: (36)(1) 392-26-92 Fax: (36)(1) 275-49-96 Fax/phone: (36)(1) 395-92-32 home page: www.mfa.kfki.hu/~labar
Yes, please forgive me. My spelling is not very accurate when I am responding so early in the morning. Your correction on Wehnelt is correct. Please forgive any such errors in this response, as it is similar in timing.
The electric field of interest is not between any parts of the column, but rather between the electrons and the sample. I really don't want to try to attempt any quantum exercises here, but the energy impressed on the electrons results in their increased EM field effects. The acceleration of the electrons controlled by the electron gun configuration controls the energy afforded the electrons. What happens after the anode does not affect the energy of the electrons in the beam.
The electrons are the field. The acceleration given to them by the gun determine their acceleration, and thus their field and relativistic effects. Their energy is determined by the fields in the gun structure alone. Once they are given that energy, their travel through the column is determined by the momentum they have been given. The fields they generate as they travel through the sample surface are the direct result of the energy they were provided with in the electron gun.
Here's a simple challenge - define the 'kinetic' energy of a fast moving electron. As the kinetic energy is generally defined as the mass vs. the velocity of that mass, you may have a problem. The best high energy experiments have been unable to assign a 'mass' to the electron. Apparently, the only mass associated with the electron is the Einsteinium energy-mass equivalent of its charge. If there is any increase in an electron's mass/energy by acceleration in an applied electric field, it is to the electrons energy. Check with SLAC's experiments.
A fine point, but one with merit. It doesn't matter whether the distance from anode to sample is 10 cm or 1000 cm, if the electron can travel the distance without interference and external force. Thus the electro-magn etic interaction of the electron with the sample is simply dependant on the force originally impressed on the electron by the gun.
The field present at the sample surface will thus be determined by the acceleration given the electrons by the gun and their current density. The higher the beam current, the greater the field flux. The smaller the area of the electron impingement on the surface, the higher the field flux. As we work towards smaller circuit dimensions, these electron interactions produce higher field flux densities as the circuit dimensions come closer to the electron beam diameters.
You seem to want to separate the mass related kinetic effects from the energy of the particle, but that can not be done. As the electron is accelerated, its energy is apparently increased, rather than its mass. The result is that the classical and relativistic effects normally related to mass are instead seen in an increase in the energy, and thus, the field effects of the electron with the sample.
On Friday, January 19, 2001 10:43 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] wrote: } { { File: ATT00000.txt; charset = windows-1252 } }
Thanks, Allen, for pointing out some of the issues here. By the way, It's "Wehnelt", not "Wienault".
You are probably right about the ESD vs. EDS name. However, it was clear to me what the original question was about.
Someone pointed out, that the cause of ESD (!) failure in electronic devices is, that the fields increase to beyond the breakdown strength of the devices, which causes a high and sudden current to flow through the parts where the fields are highest, which in turn heats and destroys the sample. I think, that is right. It's the fields that reach critical strength, which then causes the current flow and destruction of the device. That's what I meant when I said you get "zapped".
Now, about the electron microscope. Obviously I don't want to pronounce, that electron microscopy is impossible. That would be stupid, wouldn't it? However, as you said yourself, the sample is at ground potential, or very near it, and so is the anode. In other words, there is not electric potential (or very little) between the anode and the sample and thus no electric field. Since the sample is grounded, as is the rest of the microscope, there is no electric field between those, either. The energy of an electron in an electric field is proportional to the electric field (actually proportional to the square of the field, if my memory serves me). No field - no electric energy. Of course that does not mean that the electrons don't have energy. They have quite a lot of energy, on the order of 20keV, by "falling" through the electric field between cathode and anode and not being stopped by the anode. Since the sample is grounded (or should be), the electric effects of the electron beam on the grounded sample will be cancelled. What you have to contend with is the kinetic energy (20 keV/electron), which is transferred to the sample.
Now, I have always talked about grounded samples. That does not mean that electric fields cannot be generated by the electron beam within the sample. Obviously, you put electrons in the sample and they have to move out of the sample. If there is any kind of resistance (an oxide layer, etc.), that will result in charging and an electric potential.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Allen R. Sampson [mailto:ars-at-sem.com] Sent: Friday, January 19, 2001 2:44 AM To: 'Mike Bode'
Apologies in advance if this has been asked before ..
Has anyone had an success in interfacing an Epson PhotoPC 3000 digital camera to an optical microscope?
Thanks
Giles Sanders ---------------------------------------------------------------------------- --------------------------- Dr. Giles Sanders Zeneca / SmithKline Beecham Centre for Analytical Sciences Chemistry Department Imperial College of Science, Technology and Medicine London SW7 2AY
Hello everybody My questions are: 1- Is it very important to use an extremly pure ethanol (AA) to remove the xylene from the semithin sections placed on slides? 2- Can the tissue burst and the cells disrupt if it is not 100% pure? 3- Can the xylene stay within the tissue if the AA is not 100% pure, as little yellow bubbles? 4- What happens if the paraffin has been stored at 56 degrees for too long? Can it turn into longer polymers and then be harder or impossible to remove from the sections? How may I detect so in the sections, is it visible?
Albert Cardona University of Barcelona, Spain.
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Just thought I'd provide a corrected phone number & address (having been to their lab). These were pulled off their website, but I didn't check to see if they have what you're looking for.
Joseph D. Geller Email: jg-at-gellermicro.com www.gellermicro.com
Geller MicroAnalytical Laboratory 426e Boston St. Topsfield, MA 01983-1212
Phone:(978) 887-7000 Fax:(978) 887-6671
Hope this helps. Disclaimer - I have no connection with Geller MicroAnalytical Laboratory except as a satisfied customer. - Louise
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-339-7300 phone 508-339-4996 fax Louise_Harner-at-albint.com
----- Forwarded by Louise Harner/AlbInt on 2001/01/22 09:32 AM -----
"Ekstrom, Harry" {harry.ekstrom-at-hone To: '\"Jim_Koncki {"'\"Jim_Koncki-at-Ingersoll-Rand.com\" ywell.com} -at-sparc5.microscopy.com'" {"Jim_Koncki-at-Ingersoll-Rand.com" -at-sparc5.microscopy.com} , Microscopy-at-sparc5.microscopy.com 2001/01/21 12:21 PM cc: Subject: RE: Carbon in Steel Standards for WDX
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You might try Geller Microanalytical Lab in Peabody, Mass. The last phone number I had for him was 508-535-5595. If he doesn't have it, he would know where to direct you.
-----Original Message----- } From: "Jim_Koncki-at-Ingersoll-Rand.com"-at-sparc5.microscopy.com [mailto:"Jim_Koncki-at-Ingersoll-Rand.com"-at-sparc5.microscopy.com] Sent: Friday, January 19, 2001 5:14 PM To: Microscopy-at-sparc5.microscopy.com
Hi All:
I need to obtain some certified standards for carbon in steel matrix to do quantitative work using WDX. The problem is finding material that is homogeneous on a micro-scale. I am trying to cover a range of 0.1 to 2.0 Wt. % carbon. No luck thus far with NIST or Brammer.
Does anyone know of a supplier who can produce and sell such material?
Thanks in advance.
Jim Koncki Senior Project Chemist Torrington Co./Ingersoll-Rand
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Dear János, I did a lot of TEM work of Mg-based alloys and I never experienced any danger of these alloys in the microscope. The Mg matrix is definitely stable in the microscope under the beam. The only concern may arise from the stability of the compounds in your samples (usually I don't think it's a big deal, since once a particle decomposes under the beam, the amount is quite tiny). Anyway you may test your sample out of the microscope first. Best regards, Z.P. Luo
} From: "Labar Janos" {labar-at-mfa.kfki.hu} } To: "Microscopy ListServer" {Microscopy-at-sparc5.microscopy.com} } Subject: TEM of Mg-based alloys } Date: Mon, 22 Jan 2001 10:41:58 +0100 } } } Dear Colleagues, } } A friend wants to examine Mg-based alloys in TEM and warned us that in a } previous lab this alloy was expelled from the microscope because it caused } contamination by sublimation. I have no experience with these materials. Can } anyone give me more detailed info if this is a real danger? } } Thanks in advance: } } János } } } Dr. habil, Janos L. Labar } Research Institute for Technical Physics and Materials Science } H-1121 Budapest, Konkoly-Thege u. 29-33 } Postal address: H-1525 Budapest-114, Po Box 49 } Tel: (36)(1) 392-26-92 } Fax: (36)(1) 275-49-96 } Fax/phone: (36)(1) 395-92-32 } home page: www.mfa.kfki.hu/~labar }
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id KAA27753 for dist-Microscopy; Mon, 22 Jan 2001 10:14:28 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id KAA27746 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 22 Jan 2001 10:13:57 -0600 (CST) Received: from e3.ny.us.ibm.com (e3.ny.us.ibm.com [32.97.182.103]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id KAA27739 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 22 Jan 2001 10:13:46 -0600 (CST) Received: from northrelay02.pok.ibm.com (northrelay02.pok.ibm.com [9.117.200.22]) by e3.ny.us.ibm.com (8.9.3/8.9.3) with ESMTP id LAA167736 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 22 Jan 2001 11:11:17 -0500 Received: from d01ml068.pok.ibm.com (d01ml068.pok.ibm.com [9.117.250.68]) by northrelay02.pok.ibm.com (8.8.8m3/NCO v4.95) with ESMTP id LAA30456 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 22 Jan 2001 11:09:11 -0500 Importance: Normal
Dear Cindy,
The following is our supplier for clear "glassine " envelopes:
National Graphic Supply 226 N. Allen Street Albany, NY 12206 Phone: (800)-223-7130
The manufacturer is Savage Universal Corporation in Arizona. We have purchased, in the past, the following sizes:
2 3/4" X 3 3/4" Part No. SA 64 OPEN END (1000 per carton) - $ 63.00 per carton* 4 1/4" X 5 1/4" Part No. SA 66 OPEN END (1000 per carton) - $ 49.40 per carton** 5 1/4" X 7 3/8" Part No. SA 67 OPEN END (1000 per carton) - $111.45 per carton* 8 1/4" X 10 1/4" Part No. SA 68 OPEN END (500 per carton) - $102.90 per carton*
Here we use the LKB plastic knife boats for our glass knives on cutting 1 micron sections. To attach the knife boat to the glass I use clear nail polish. The problem is how can I safely remove the nail polish and use the knife boats again.
I have tried a low concentration of acetone which melts the boats. I have tried a diluted nail polish remover which also melts the plastic boats and deforms them.
Any suggestions?
Thanks,
Eric A. Rosen UCLA Medical Center Electron Microscopy Lab Department of Pathology Email: biology-at-ucla.edu or erosen-at-mednet.ucla.edu
} I was wondering if there are any stains/methods that would allow the } identification of basal bodies or centromeres in a non-dividing cell. } I would like to determine the basal body position in a cell without } resorting to serial TEM. My thinking is that I might be able to do } it in whole cells with the right marker. Any comments?
This is classical cytology, so you need a classical method! Here are a few suggestions.
1. Osmium tetroxide is a must, either in the primary fixative or as a post-fix. A glutaraldehyde-osmium sequence, as for EM, is excellent.
2. Plastic embedding followed by staining 1um sections with a basic dye may be all you need. Otherwise, cut paraffin sections as thinly as possible (The 19th century methods for centrosomes were, according to McClung's Handbook, done this way, after chrome-osmic fixatives.)
3. Try treating paraffin sections of postosmicated tissue with ethyl gallate, which makes a more darkly coloured product at sites of osmium binding. This method was used with great elegance in a study of the hypothalamus and neurohypophysis by D. G. Montemurro - J. Endocrinol. 35: 271-279 (1966).
4. Try Heidenhain's iron-haematoxylin method. With careful differentiation this can make almost anything visible.
5. When you have found a good method let us all know about it, by way of Histonet.
John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1 E-mail: kiernan-at-uwo.ca http://publish.uwo.ca/~jkiernan/index.htm
I didn't think that you could get a single-phase microstructure for carbon in steel at a very high level of carbon. But I couldn't remember with certainty, so I asked my friendly neighborhood materials scientist. Here is his reply. ------------- } At what levels can carbon be held in the matrix by solid solution? When } does it force a second phase to appear?
About 0.02% C.
That's a huge range of carbon levels to expect homogenous microstructure. Only thing I can think of would be to look for standards with nickel to keep the steel austenitic, like an austenitic stainless steel. Might be able to get quenched steel standards having a somewhat uniform structure of martensite, though not purely homogenous.
So it begs the question, if the guy needs standards to analyze samples that are homogenous, how are the samples being prepared? ------------- I would be curious to know what sort of samples will be the subject of the investigation. I suppose it might be possible to get some estimate by probing a larger area, but the results would be subject to dis-similarities in the microstructure and whatever errors result from dealing with a two-phase structure.
C in iron is problematic, and I don't have a probe to get enough signal. Besides, my friend gets good, cheap, fast results from a spectrographic lab so that I haven't bothered to even try.
Warren S.
At 06:14 PM 1/19/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ok, here I go again :-) We've checked the PBS pH and turned out to be just fine, about 7 (suposedly is 7.2). The glycerol was more acid though, down to 4.5. So maybe it was the glycerol. But we performed a dot-blot test and it was negative for that batch of glycerol: we fixed HRPeroxidase on nitrocelulose paper and we tested 2 batches of DAB (an old and a new one)to reveal it, and both performed equally up to the same HRP-dilution. Then we added the supposedly bad glycerol:PBS and nothing happened, the stain didn't fade, not even in 2 hours. So that is why it's running us crazy. Maybe the DAB performs differently on semithin tissue sections than over nitrocellulose paper. The semithin sections, well they are 7 or 10 micrometers wide,in 53ª paraffin, and a few months ago the immunostaining didn't work at all (though the protocol has been working fine for 5 years), and then small yellowish bubbles could be seen inside the tissue. And we never cleared if they were residual paraffin or xylene -though we found out both paraffin and xylene were doing wrong, since the paraffin had been at 56º for months -supposedly the polymers grew longer-, the xylene was too polluted with paraffin and the AA was not pure at all. I'll never forget again that EVERY single step on that protocol is killingly important. That is why I was asking if anyone had ever had any problems with immunostainings ... So, what do you think? Right now the protocol works fine some days, and some others doesn't.
Albert Cardona University of Barcelona
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Just thought I would throw my two cents worth in regarding the issue of fields. It seems that one of the problems is defining exactly what we mean by an electric field. Faraday introduced the concept of a field (or more precisely a "force field") simply as model or construct used to describe the existence - i.e. the observation - of electric forces. An electric field is characterized or explored by "measuring" the magnitude and direction of electric force experienced by a test charge (the force vector). A collection of infinitely many force vectors "defines" the electric field. At any point in the field, the direction of the field is DEFINED as the direction of force on the test charge, and the strength of the field is DEFINED as the magnitude of the force experienced by the test charge. There are different models for the mechanism of electric force - the "original" Faraday-Maxwell theory, in which lines or tubes of force represent a strain in the "ether" - to the more modern theory of exchange of particles (photon-field theory). But the important point here is that "field" is simply a concept used to characterize the electric forces experienced by our imaginary test charge (we'll say an electron, even though physicists use a positive test charge by convention).
So maybe the question to ask is what electric force will an imaginary, stationary electron experience in the "vicinity" of the sample (either above the sample surface or within the sample). Assuming that the sample is a perfect conductor, my sense is that this stationary electron would not experience any significant electric forces, indicating that (by definition) no significant electric field exists.
However, real samples such as what we're discussing in this thread will contain regions of varying electrical conductivity. The best example is perhaps that of a charging dust particle, around which a strong electrical "field" is generated - I believe a stationary test charge will experience strong electrical forces in this region. The reason the field is produced is the existence of a buildup of charges on/in the dust particle, and the close proximity of the surface of the sample which does not build up this electric charge (perhaps semantics, but I would NOT say the electrons ARE the field - they may be partly responsible for the field, but the "field" itself is just a concept which characterizes our observation that a test charge in this region will experience a force which has magnitude and direction.
Therefore, it seems to me that any electrostatic discharge (ESD) damage caused within a sample will be initiated by the generation of such an electric field, which in turn can only be generated when there are differences in electrical conductivity (otherwise, how can a differential buildup of charges - which produces the field - occur?)
Best regards,
Tony mailto:towens-at-camscan-usa.com
Tony Owens CamScan USA Inc. 508 Thomson Park Dr. Cranberry Twp., PA 16066 Tel: 724-772-7433 Fax: 724-772-7434 URL: www.camscan-usa.com
____________________________________________
Monday, January 22, 2001, 1:44:53 PM, You wrote:
ARS} ------------------------------------------------------------------------ ARS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America ARS} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com ARS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ARS} -----------------------------------------------------------------------.
ARS} Yes, please forgive me. My spelling is not very accurate when I am ARS} responding so early in the morning. Your correction on Wehnelt is correct. ARS} Please forgive any such errors in this response, as it is similar in ARS} timing.
ARS} The electric field of interest is not between any parts of the column, but ARS} rather between the electrons and the sample. I really don't want to try to ARS} attempt any quantum exercises here, but the energy impressed on the ARS} electrons results in their increased EM field effects. The acceleration of ARS} the electrons controlled by the electron gun configuration controls the ARS} energy afforded the electrons. What happens after the anode does not ARS} affect the energy of the electrons in the beam.
ARS} The electrons are the field. The acceleration given to them by the gun ARS} determine their acceleration, and thus their field and relativistic ARS} effects. Their energy is determined by the fields in the gun structure ARS} alone. Once they are given that energy, their travel through the column is ARS} determined by the momentum they have been given. The fields they generate ARS} as they travel through the sample surface are the direct result of the ARS} energy they were provided with in the electron gun.
ARS} Here's a simple challenge - define the 'kinetic' energy of a fast moving ARS} electron. As the kinetic energy is generally defined as the mass vs. the ARS} velocity of that mass, you may have a problem. The best high energy ARS} experiments have been unable to assign a 'mass' to the electron. ARS} Apparently, the only mass associated with the electron is the Einsteinium ARS} energy-mass equivalent of its charge. If there is any increase in an ARS} electron's mass/energy by acceleration in an applied electric field, it is ARS} to the electrons energy. Check with SLAC's experiments.
ARS} A fine point, but one with merit. It doesn't matter whether the distance ARS} from anode to sample is 10 cm or 1000 cm, if the electron can travel the ARS} distance without interference and external force. Thus the electro-magn ARS} etic interaction of the electron with the sample is simply dependant on the ARS} force originally impressed on the electron by the gun.
ARS} The field present at the sample surface will thus be determined by the ARS} acceleration given the electrons by the gun and their current density. The ARS} higher the beam current, the greater the field flux. The smaller the area ARS} of the electron impingement on the surface, the higher the field flux. As ARS} we work towards smaller circuit dimensions, these electron interactions ARS} produce higher field flux densities as the circuit dimensions come closer ARS} to the electron beam diameters.
ARS} You seem to want to separate the mass related kinetic effects from the ARS} energy of the particle, but that can not be done. As the electron is ARS} accelerated, its energy is apparently increased, rather than its mass. The ARS} result is that the classical and relativistic effects normally related to ARS} mass are instead seen in an increase in the energy, and thus, the field ARS} effects of the electron with the sample.
ARS} On Friday, January 19, 2001 10:43 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] ARS} wrote: } } { { File: ATT00000.txt; charset = windows-1252 } }
ARS} Thanks, Allen, for pointing out some of the issues here. By the way, It's ARS} "Wehnelt", not "Wienault".
ARS} You are probably right about the ESD vs. EDS name. However, it was clear to ARS} me what the original question was about.
ARS} Someone pointed out, that the cause of ESD (!) failure in electronic ARS} devices ARS} is, that the fields increase to beyond the breakdown strength of the ARS} devices, which causes a high and sudden current to flow through the parts ARS} where the fields are highest, which in turn heats and destroys the sample. ARS} I ARS} think, that is right. It's the fields that reach critical strength, which ARS} then causes the current flow and destruction of the device. That's what I ARS} meant when I said you get "zapped".
ARS} Now, about the electron microscope. Obviously I don't want to pronounce, ARS} that electron microscopy is impossible. That would be stupid, wouldn't it? ARS} However, as you said yourself, the sample is at ground potential, or very ARS} near it, and so is the anode. In other words, there is not electric ARS} potential (or very little) between the anode and the sample and thus no ARS} electric field. Since the sample is grounded, as is the rest of the ARS} microscope, there is no electric field between those, either. The energy of ARS} an electron in an electric field is proportional to the electric field ARS} (actually proportional to the square of the field, if my memory serves me). ARS} No field - no electric energy. Of course that does not mean that the ARS} electrons don't have energy. They have quite a lot of energy, on the order ARS} of 20keV, by "falling" through the electric field between cathode and anode ARS} and not being stopped by the anode. Since the sample is grounded (or should ARS} be), the electric effects of the electron beam on the grounded sample will ARS} be cancelled. What you have to contend with is the kinetic energy (20 ARS} keV/electron), which is transferred to the sample.
ARS} Now, I have always talked about grounded samples. That does not mean that ARS} electric fields cannot be generated by the electron beam within the sample. ARS} Obviously, you put electrons in the sample and they have to move out of the ARS} sample. If there is any kind of resistance (an oxide layer, etc.), that ARS} will ARS} result in charging and an electric potential.
ARS} -----Original Message----- } } From: Allen R. Sampson [mailto:ars-at-sem.com] ARS} Sent: Friday, January 19, 2001 2:44 AM ARS} To: 'Mike Bode' ARS} Subject: RE: Question
ARS} Ok, apparently a simple physics lesson needed here.
ARS} First of all we are talking about ESD, or Electro-Static Discharge effects ARS} here. Not EDS, or what in this field is known as Electron Dispersive ARS} Spectroscopy.
ARS} Secondly, the electrons arriving at the sample surface in an SEM are ARS} getting there with an acceleration that is roughly equivalent to the the ARS} acceleration voltage impressed at the cathode. The sample is at, or very ARS} near, ground potential. But the electrons in the beam are reaching the ARS} sample at close to the accelerating voltage impressed. The grid or ARS} "Wienault" of the electron gun is at a potential roughly +500V from the ARS} accelerating voltage and is used to induce the electrons from the cathode ARS} through the aperture in the anode, preventing the 'space charge' effects ARS} around the cathode and providing the first electrostatic lens in the gun.
ARS} The potential of the anode of an electron microscope may be at ground ARS} potential, but the electron beam is passing through the opening in the ARS} anode. The anode is used as the second electrostatic lens, and has little ARS} effect on the energy of the electrons passing through it. By its charge ARS} opposite that of the electron beam, it encourages the first crossover of ARS} the beam, but since the beam does not directly interact with it, there is ARS} no direct energy exchange.
ARS} Precisely what kinetic energy can be transferred to the sample if there is ARS} no energy differential between the electron potential at the anode and the ARS} sample? Given your understanding of the processes involved, there can be ARS} no discernable energy changes in the sample since the electrons have no ARS} energy, and thus, there can be no discernable energy release from the ARS} sample. In other words, in your view, electron microscopy is impossible.
ARS} On Thursday, January 18, 2001 2:24 PM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] ARS} wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Yes, EDS is very real and chip manufacturers go to great length (end } } expense) to avoid them. However, I think the mechanism is different: } } } } EDS means, that something collects static electricity (by walking on a } } carpet, for example), then touches something that is at a very different } } potential. That's what happens if you touch a metal after walking on that } } carpet. The enormous potential difference create strong electric field ARS} that } } finally ionize the air and you get "zapped". The strong fields can ARS} destroy } } the electronics. } } } } In an SEM the sample is usually grounded and does not see strong fields. } } True, the electrons are accelerated to 30 KV or more, but the sample does } } not see that, as the anode usually sits at ground level with the Cathode } } being at - 30 kV. What you need to be concerned about here is the kinetic } } energy that is transferred to the sample and can heat it up considerable. ARS} On } } the other hand, if the sample is not grounded, there will be fields } } developing. There are also other effects, such as residual organics ARS} getting } } "cracked" and forming a residue on the sample, but that's a different ARS} story. } } } } Michael } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } phone: (888) FIND SIS } } (303) 234-9270 } } fax: (303) 234-9271 } } email: mailto:info-at-soft-imaging.com } } web: http://www.soft-imaging.com } } =================================== } } ARS} Allen R. Sampson, Owner ARS} Advanced Research Systems ARS} 317 North 4th. Street ARS} St. Charles, Illinois 60174 ARS} voice 630.513.7093 fax 630.513.7092
ARS} Allen R. Sampson, Owner ARS} Advanced Research Systems ARS} 317 North 4th. Street ARS} St. Charles, Illinois 60174 ARS} voice 630.513.7093 fax 630.513.7092
Hiyo Listers, What are the problems associated with imaging (SE and BSE) low level radioactive samples (rocks)? They were irradiated for isotopic dating ~2 years ago and are now only a couple of times background? Does anyone have experience with this low level radioactivity and SEM analysis? I would appreciate any advice. Many thanks, Sarah
-- Sarah A.W. Lundberg Electron Microanalysis and Imaging Laboratory Department of Geoscience, UNLV 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010
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} Hiyo Listers, } What are the problems associated with imaging (SE and BSE) low level } radioactive samples (rocks)? They were irradiated for isotopic dating } ~2 years ago and are now only a couple of times background? Does anyone } have experience with this low level radioactivity and SEM analysis? I } would appreciate any advice. } Many thanks, } Sarah } } -- } Sarah A.W. Lundberg
Sarah,
From your description there should not be any problem with SE and BSE imaging if the levels are only a few times "background".
If your samples have been irradiated (neutrons?) at some point in the past then you might pick up some spontaneous signals from them in your EDS detector, but that's probably about the extent of any interference you'll get.
Cheers, Arthur.
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
The concept of fields are not, I think, as arbitrary as you seem to believe. Their introduction was not as an expedient to describe electric forces, but rather DEFINED as their influence over distance, which could not be explained by any other means. A fine point, perhaps, but one worth making. Newton had given some explanation of the gravitational field, and thus the underpinning of field theory. When it was discovered that electricity and magnetism were related, it was obvious that the same theories that explained the mass-gravity duality might be extended to charge-magnetism . In each case, it was supposed that distant interactions could be related to localized observations.
In our specific field, we are privy to the roots of quantum theory that currently pervade our understanding of this world. The characteristic x-ray spectrums that we rely on were a catalyst for the basic quantum theories such as Pauli's, not to mention the photo-electric effects of PMTs that were the foundation of Einstein's theory of Special Relativity. These theories also support the concept of fields, although they couch them in the exchange of discrete particles. This is used to explain the empirically observed discrete, rather than continuous, exchanges of energy between sub-atomic particles.
You digress to interactions of a sample with 'stationary' electrons. I have written about electrons that have been given energy by acceleration in electric fields. Apples and oranges. An electron that has been accelerated by the typical electron beam instrument has far more energy than a 'stationary' electron, and that energy is primarily available through its interaction with matter. Can a 'stationary' electron produce an inner shell electron vacancy? I can prove that an accelerated electron can produce far greater yields than those produced by simple kinetic theories. Producing an inner shell electron vacancy requires a defined amount of energy and energy transfer at a distance that is simply not available in a 'stationary' electron. As the electron has no measurable dimension, the 'billiard ball' idea of kinetics can simply not explain such an energy exchange or the spectral yield in a typical x-ray instrument.
A simple demonstration. Within your SEM or TEM is a circuit that provides compensation for various accelerating voltages used. When you increase the accelerating voltage, this circuit will increase the currents applied to the condenser and objective lenses. Why? Because the electrons have more energy at the increased accelerating voltages and require more force to be applied to alter their trajectories to produce a similar demagnification and focussing. The rest energy of these electrons has not changed, but their total energy has. What repercussions does this have? Very simply, their ability to interact with external fields and thus other charged particles has increased while their susceptibility to those influences has decreased.
In terms of fields, the accelerated electron produces an increased electric field gradient in its immediate vicinity as its energy is increased. For a single electron, this is a very localized effect. But in an electron beam instrument, this effect can be spread over a much larger area as there is considerable uncertainty in the location of individual electrons. In a macro gradient produced by ESD, they are larger still. While the field gradients are substantially different from those produced by electro-static discharge, the effect and cause can be basically the same.
In ESD, two macro materials are brought in close contact that have greatly differing electric potentials. Those electric differentials eventually find a path through ionization of the interstitial gases to equalize the electrical difference. The intervening gases provide a randomizing effect that spreads the effect over a relatively large area. However, the energy transfer behind the effect is the acceleration of electrons by the differing electric potentials of the materials. Those accelerated electrons hit the secondary material with the same enhanced energy as those accelerated by artificial means.
Whether you use particle energy and fields or wave functions, the math is similar. The electron has a rest mass, that is indistinguishable from its charge. When accelerated through electric fields, the electron gains energy that is then available externally, through interactions with the fields it generates, or the extension of its wave function. In either case, as the energy of the electron is increased, the gradient of its field or wave function increases. That creates an increasing EM field that on a small scale can create large electric and magnetic fields gradients that can emulate larger scale effects within reason.
On Monday, January 22, 2001 1:39 PM, Tony Owens [SMTP:towens-at-camscan-usa.com] wrote: } Just thought I would throw my two cents worth in regarding the } issue of fields. It seems that one of the problems is defining } exactly what we mean by an electric field. Faraday introduced the } concept of a field (or more precisely a "force field") simply as } model or construct used to describe the existence - i.e. the } observation - of electric forces. An electric field is } characterized or explored by "measuring" the magnitude and } direction of electric force experienced by a test charge (the } force vector). A collection of infinitely many force vectors } "defines" the electric field. At any point in the field, the } direction of the field is DEFINED as the direction of force on } the test charge, and the strength of the field is DEFINED as the } magnitude of the force experienced by the test charge. There are } different models for the mechanism of electric force - the } "original" Faraday-Maxwell theory, in which lines or tubes of } force represent a strain in the "ether" - to the more modern } theory of exchange of particles (photon-field theory). But the } important point here is that "field" is simply a concept used to } characterize the electric forces experienced by our imaginary } test charge (we'll say an electron, even though physicists use a } positive test charge by convention). } } So maybe the question to ask is what electric force will an } imaginary, stationary electron experience in the "vicinity" of } the sample (either above the sample surface or within the } sample). Assuming that the sample is a perfect conductor, my } sense is that this stationary electron would not experience any } significant electric forces, indicating that (by definition) no } significant electric field exists. } } However, real samples such as what we're discussing in this } thread will contain regions of varying electrical conductivity. } The best example is perhaps that of a charging dust particle, } around which a strong electrical "field" is generated - I believe } a stationary test charge will experience strong electrical forces } in this region. The reason the field is produced is the existence } of a buildup of charges on/in the dust particle, and the } close proximity of the surface of the sample which does not build } up this electric charge (perhaps semantics, but I would NOT say } the electrons ARE the field - they may be partly responsible for the } field, but the "field" itself is just a concept which } characterizes our observation that a test charge in this region } will experience a force which has magnitude and direction. } } Therefore, it seems to me that any electrostatic discharge (ESD) } damage caused within a sample will be initiated by the generation } of such an electric field, which in turn can only be generated } when there are differences in electrical conductivity (otherwise, } how can a differential buildup of charges - which produces the } field - occur?) } } Best regards, } } } Tony mailto:towens-at-camscan-usa.com } } } Monday, January 22, 2001, 1:44:53 PM, You wrote: } } ARS} Yes, please forgive me. My spelling is not very accurate when I am } ARS} responding so early in the morning. Your correction on Wehnelt is correct. } ARS} Please forgive any such errors in this response, as it is similar in } ARS} timing. } } ARS} The electric field of interest is not between any parts of the column, but } ARS} rather between the electrons and the sample. I really don't want to try to } ARS} attempt any quantum exercises here, but the energy impressed on the } ARS} electrons results in their increased EM field effects. The acceleration of } ARS} the electrons controlled by the electron gun configuration controls the } ARS} energy afforded the electrons. What happens after the anode does not } ARS} affect the energy of the electrons in the beam. } } ARS} The electrons are the field. The acceleration given to them by the gun } ARS} determine their acceleration, and thus their field and relativistic } ARS} effects. Their energy is determined by the fields in the gun structure } ARS} alone. Once they are given that energy, their travel through the column is } ARS} determined by the momentum they have been given. The fields they generate } ARS} as they travel through the sample surface are the direct result of the } ARS} energy they were provided with in the electron gun. } } ARS} Here's a simple challenge - define the 'kinetic' energy of a fast moving } ARS} electron. As the kinetic energy is generally defined as the mass vs. the } ARS} velocity of that mass, you may have a problem. The best high energy } ARS} experiments have been unable to assign a 'mass' to the electron. } ARS} Apparently, the only mass associated with the electron is the Einsteinium } ARS} energy-mass equivalent of its charge. If there is any increase in an } ARS} electron's mass/energy by acceleration in an applied electric field, it is } ARS} to the electrons energy. Check with SLAC's experiments. } } ARS} A fine point, but one with merit. It doesn't matter whether the distance } ARS} from anode to sample is 10 cm or 1000 cm, if the electron can travel the } ARS} distance without interference and external force. Thus the electro-magn } ARS} etic interaction of the electron with the sample is simply dependant on the } ARS} force originally impressed on the electron by the gun. } } ARS} The field present at the sample surface will thus be determined by the } ARS} acceleration given the electrons by the gun and their current density. The } ARS} higher the beam current, the greater the field flux. The smaller the area } ARS} of the electron impingement on the surface, the higher the field flux. As } ARS} we work towards smaller circuit dimensions, these electron interactions } ARS} produce higher field flux densities as the circuit dimensions come closer } ARS} to the electron beam diameters. } } ARS} You seem to want to separate the mass related kinetic effects from the } ARS} energy of the particle, but that can not be done. As the electron is } ARS} accelerated, its energy is apparently increased, rather than its mass. The } ARS} result is that the classical and relativistic effects normally related to } ARS} mass are instead seen in an increase in the energy, and thus, the field } ARS} effects of the electron with the sample. } } ARS} On Friday, January 19, 2001 10:43 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] } ARS} wrote: } } } { { File: ATT00000.txt; charset = windows-1252 } } } } ARS} Thanks, Allen, for pointing out some of the issues here. By the way, It's } ARS} "Wehnelt", not "Wienault". } } ARS} You are probably right about the ESD vs. EDS name. However, it was clear to } ARS} me what the original question was about. } } ARS} Someone pointed out, that the cause of ESD (!) failure in electronic } ARS} devices } ARS} is, that the fields increase to beyond the breakdown strength of the } ARS} devices, which causes a high and sudden current to flow through the parts } ARS} where the fields are highest, which in turn heats and destroys the sample. } ARS} I } ARS} think, that is right. It's the fields that reach critical strength, which } ARS} then causes the current flow and destruction of the device. That's what I } ARS} meant when I said you get "zapped". } } ARS} Now, about the electron microscope. Obviously I don't want to pronounce, } ARS} that electron microscopy is impossible. That would be stupid, wouldn't it? } ARS} However, as you said yourself, the sample is at ground potential, or very } ARS} near it, and so is the anode. In other words, there is not electric } ARS} potential (or very little) between the anode and the sample and thus no } ARS} electric field. Since the sample is grounded, as is the rest of the } ARS} microscope, there is no electric field between those, either. The energy of } ARS} an electron in an electric field is proportional to the electric field } ARS} (actually proportional to the square of the field, if my memory serves me). } ARS} No field - no electric energy. Of course that does not mean that the } ARS} electrons don't have energy. They have quite a lot of energy, on the order } ARS} of 20keV, by "falling" through the electric field between cathode and anode } ARS} and not being stopped by the anode. Since the sample is grounded (or should } ARS} be), the electric effects of the electron beam on the grounded sample will } ARS} be cancelled. What you have to contend with is the kinetic energy (20 } ARS} keV/electron), which is transferred to the sample. } } ARS} Now, I have always talked about grounded samples. That does not mean that } ARS} electric fields cannot be generated by the electron beam within the sample. } ARS} Obviously, you put electrons in the sample and they have to move out of the } ARS} sample. If there is any kind of resistance (an oxide layer, etc.), that } ARS} will } ARS} result in charging and an electric potential. } } } ARS} -----Original Message----- } } } From: Allen R. Sampson [mailto:ars-at-sem.com] } ARS} Sent: Friday, January 19, 2001 2:44 AM } ARS} To: 'Mike Bode' } ARS} Subject: RE: Question } } } ARS} Ok, apparently a simple physics lesson needed here. } } ARS} First of all we are talking about ESD, or Electro-Static Discharge effects } ARS} here. Not EDS, or what in this field is known as Electron Dispersive } ARS} Spectroscopy. } } ARS} Secondly, the electrons arriving at the sample surface in an SEM are } ARS} getting there with an acceleration that is roughly equivalent to the the } ARS} acceleration voltage impressed at the cathode. The sample is at, or very } ARS} near, ground potential. But the electrons in the beam are reaching the } ARS} sample at close to the accelerating voltage impressed. The grid or } ARS} "Wienault" of the electron gun is at a potential roughly +500V from the } ARS} accelerating voltage and is used to induce the electrons from the cathode } ARS} through the aperture in the anode, preventing the 'space charge' effects } ARS} around the cathode and providing the first electrostatic lens in the gun. } } ARS} The potential of the anode of an electron microscope may be at ground } ARS} potential, but the electron beam is passing through the opening in the } ARS} anode. The anode is used as the second electrostatic lens, and has little } ARS} effect on the energy of the electrons passing through it. By its charge } ARS} opposite that of the electron beam, it encourages the first crossover of } ARS} the beam, but since the beam does not directly interact with it, there is } ARS} no direct energy exchange. } } ARS} Precisely what kinetic energy can be transferred to the sample if there is } ARS} no energy differential between the electron potential at the anode and the } ARS} sample? Given your understanding of the processes involved, there can be } ARS} no discernable energy changes in the sample since the electrons have no } ARS} energy, and thus, there can be no discernable energy release from the } ARS} sample. In other words, in your view, electron microscopy is impossible. } } } } ARS} On Thursday, January 18, 2001 2:24 PM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] } ARS} wrote: } } } } } } } } } Yes, EDS is very real and chip manufacturers go to great length (end } } } expense) to avoid them. However, I think the mechanism is different: } } } } } } EDS means, that something collects static electricity (by walking on a } } } carpet, for example), then touches something that is at a very different } } } potential. That's what happens if you touch a metal after walking on that } } } carpet. The enormous potential difference create strong electric field } ARS} that } } } finally ionize the air and you get "zapped". The strong fields can } ARS} destroy } } } the electronics. } } } } } } In an SEM the sample is usually grounded and does not see strong fields. } } } True, the electrons are accelerated to 30 KV or more, but the sample does } } } not see that, as the anode usually sits at ground level with the Cathode } } } being at - 30 kV. What you need to be concerned about here is the kinetic } } } energy that is transferred to the sample and can heat it up considerable. } ARS} On } } } the other hand, if the sample is not grounded, there will be fields } } } developing. There are also other effects, such as residual organics } ARS} getting } } } "cracked" and forming a residue on the sample, but that's a different } ARS} story. } } } } } } Michael } } } } } } Michael Bode, Ph.D. } } } Soft Imaging System Corp. } } } 1675 Carr St., #105N } } } Lakewood, CO 80215 } } } =================================== } } } phone: (888) FIND SIS } } } (303) 234-9270 } } } fax: (303) 234-9271 } } } email: mailto:info-at-soft-imaging.com } } } web: http://www.soft-imaging.com } } } ===================================
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
You could attach the boats with wax. They can then be reused.
Dave
On Mon, 22 Jan 2001 08:22:14 -0800 ERIC {biology-at-ucla.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To my fellow Microscopists, } } Here we use the LKB plastic knife boats for our glass knives on cutting 1 } micron sections. To attach the knife boat to the glass I use clear nail } polish. The problem is how can I safely remove the nail polish and use the } knife boats again. } } I have tried a low concentration of acetone which melts the boats. I have } tried a diluted nail polish remover which also melts the plastic boats and } deforms them. } } Any suggestions? } } Thanks, } } Eric A. Rosen } UCLA Medical Center } Electron Microscopy Lab } Department of Pathology } Email: biology-at-ucla.edu or erosen-at-mednet.ucla.edu } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Solvents and polymers are my business, rather than clearing sections (I last did that sort of thing many years ago, in my schooldays), but here's some chemical background. But those with more direct experience in this field might know better.
On Mon, 22 Jan 2001, Grup Epigenetica wrote:
} Hello everybody } My questions are:
} 1- Is it very important to use an extremly pure ethanol (AA) to remove } the xylene from the semithin sections placed on slides?
I doubt it very much. DRY is important, for if water is present one might end up with xylene-water emulsion. But I seem to remember that denatured ethanol (i.e. with some methanol but not all the other things which are put into ordinary "methylated spirit) was OK.
} 2- Can the tissue burst and the cells disrupt if it is not 100% pure?
If you are finding trouble with ethanol, I would think that isopropanol (propan-2-ol) MIGHT be even better, since it has greater molar volume. But because of its greater molar volume it would work more slowly. Paint strippers, which work by swelling and bursting, rely on SMALL molar volume for their effect. Small traces of water AND of xylene tend to evaporate out of propan-2-ol, since in this environment their partial vapour pressure is greater with an azeotropic effect.
} 3- Can the xylene stay within the tissue if the AA is not 100% pure, } as little yellow bubbles?
Very unlikely, unless there is water there.
} 4- What happens if the paraffin has been stored at 56 degrees for too } long? Can it turn into longer polymers and then be harder or impossible } to remove from the sections? How may I detect so in the sections, is it } visible?
The oxidation products of paraffins are unlikely to be polymeric. Generally oxidation reduces the molecular weight of polythene, which after all is a super-long paraffin. Yellow sticky polymeric nasties only form from unsaturated oils such as linseed oil and overheated cooking oil: they can cross-link because of their double bonds. But if the paraffin is not properly washed out by the xylene it could be re-precipitated by the alcohol, whether clean or degraded.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
It is unlikely that there is a solvent that will dissolve nail polish without also attacking the boats, since the solvents in nail polish dissolve the boats. Your only option here is to use another "glue". Good old dental wax is probably the simplest.
Chris
Date sent: Mon, 22 Jan 2001 08:22:14 -0800 To: Microscopy-at-sparc5.microscopy.com } From: ERIC {biology-at-ucla.edu}
Agreed. Wax is the way to fasten boats. However, I much prefer dental wax over common paraffin wax which is more prone to cause leaky boats. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, January 23, 2001 7:53 PM, Patton, David [SMTP:David.Patton-at-uwe.ac.uk] wrote: } } } You could attach the boats with wax. They can then be } reused. } } Dave } } } On Mon, 22 Jan 2001 08:22:14 -0800 ERIC {biology-at-ucla.edu} } wrote: } } } } } } To my fellow Microscopists, } } } } Here we use the LKB plastic knife boats for our glass knives on cutting 1 } } micron sections. To attach the knife boat to the glass I use clear nail } } polish. The problem is how can I safely remove the nail polish and use the } } knife boats again. } } } } I have tried a low concentration of acetone which melts the boats. I have } } tried a diluted nail polish remover which also melts the plastic boats and } } deforms them. } } } } Any suggestions? } } } } Thanks, } } } } Eric A. Rosen } } UCLA Medical Center } } Electron Microscopy Lab } } Department of Pathology } } Email: biology-at-ucla.edu or erosen-at-mednet.ucla.edu } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" }
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Thanks Jim. I should have said it was dental wax. You can get it from an EM supply company. We have a little LKB multiplate which allows you to, warm the knife, melt the dental wax and apply it with a sort of warmed metal spatula.
Dave
On Tue, 23 Jan 2001 22:51:38 +1000 Jim at ProSciTech {jim-at-proscitech.com} wrote:
} Agreed. Wax is the way to fasten boats. However, I much prefer dental wax over } common paraffin wax which is more prone to cause leaky boats. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Tuesday, January 23, 2001 7:53 PM, Patton, David } [SMTP:David.Patton-at-uwe.ac.uk] wrote: } } } } } } You could attach the boats with wax. They can then be } } reused. } } } } Dave } } } } } } On Mon, 22 Jan 2001 08:22:14 -0800 ERIC {biology-at-ucla.edu} } } wrote: } } } } } } } } } } To my fellow Microscopists, } } } } } } Here we use the LKB plastic knife boats for our glass knives on cutting 1 } } } micron sections. To attach the knife boat to the glass I use clear nail } } } polish. The problem is how can I safely remove the nail polish and use the } } } knife boats again. } } } } } } I have tried a low concentration of acetone which melts the boats. I have } } } tried a diluted nail polish remover which also melts the plastic boats and } } } deforms them. } } } } } } Any suggestions? } } } } } } Thanks, } } } } } } Eric A. Rosen } } } UCLA Medical Center } } } Electron Microscopy Lab } } } Department of Pathology } } } Email: biology-at-ucla.edu or erosen-at-mednet.ucla.edu } } } } } } } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
At the rad levels you describe, the effects will be difficult to notice. I have examined specimens ranging up to 10s of R at a foot. Not many that hot lately, thankfully. Emissions from my specimens have included alpha, beta, and gamma. Gamma was my least favorite. Highly active alpha/beta would significantly impair imaging and EDS by adding noise to the system. The signal to noise of the SE/BSE system was degraded and EDS peaks broadened. Have you ever seen 80% dead time *before* you turn on the beam? On one occasion, the EDS was totally locked out at 100% DT. Dead time...Hummmm... OTOH those high energy gammas would escape the chamber and take aim at me. I was hiding behind two - four inches of temporary lead wall in front of the chamber.
Woody
} } ------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hiyo Listers, } What are the problems associated with imaging (SE and BSE) low level } radioactive samples (rocks)? They were irradiated for isotopic dating } ~2 years ago and are now only a couple of times background? } Does anyone } have experience with this low level radioactivity and SEM analysis? I } would appreciate any advice. } Many thanks, } Sarah } } -- } Sarah A.W. Lundberg } Electron Microanalysis and Imaging Laboratory } Department of Geoscience, UNLV } 4505 S. Maryland Parkway Box 454010 } Las Vegas, NV 89154-4010 } } EPMA Lab (702) 895-2660 } SEM Lab (702) 895-2462 } Office (702) 895-1134 } Fax (702) 895-4064 } Dept. Office (702) 895-3262 } } lundberg-at-nevada.edu } http://www.unlv.edu/Colleges/Sciences/Geoscience/EMIL.htm } } Home: } 8025 W. Russell Rd. Apt. #1117 } Las Vegas, NV 89113 } (702) 871-9635 } } }
Hello Listers, sorry but I have already deleted the original question, but I suggest "physical" cleaning by liquid nitrogen. This should also remove the nail polish.
Hope this helps
Klaus Neumann Dr. Klaus Neumann 2.5 Med. Biologie Universität des Saarlandes D-66421 Homburg Tel: ++49-6841-16-62 55 Fax: ++49-6841-16-62 56 http://www.med-rz.uni-sb.de/med_fak/biologie
Thank you Carmen Martin Yes, we do also use our dehydratation series a lot, up to 60 slides -which roughly means 2 weeks- and for sure the clearest sign is they don't get stained. But it has it all puzzling us since anything seemed to be wrong but everything failed -or so it looks.
Albert Cardona University of Barcelona. Spain.
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I have been filtered out of posting responses by a quirk of my e-mail address so this info is a little out of date but unavoidably so.
For those people with TAAB or the branded Reichert-Jung Histoknifemakers we are still able to supply many of the spare parts for instruments up to 20 years old, but don't all rush at once!
This also confirms that we are still here live and well in Blighty,
Regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England
may I just add to the discussion about the knife boats:
yes, if you use dental wax to glue the Truf to the knife (with the Leica Multiplate, especially designed for that purpose its really easy) the boat can be reused a few times.
HOWEVER: 1. breaking off the Truf from the knife and scratching away the old hardened wax may damage the Truf - leaking may result from this! Can be frustrating if you have perfect sections of a precious sample and see them dive away ... 2. Esp. with ultrathins (but also with semithins) you must be aware of possible cross contamination of old sections to the new set (I have no idea if there is a GLP rule on that but in a hospital mixing up different samples might cause serious problems)!
BTW the "LKB"Trufs are "Leica"Trufs now, but call them as you like it :-)
Best regards,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all, I just wanted to run some interesting calibration data by you. We are working with some small particles and have to have reasonable size calibrations at both 80 and 100kV on a Philips EM-400 microscope (~20 years old). Calibration using replica gratings and catalase lattice spacing give us values somewhat below the magnifications indicated by the mag. readout on the panel. This is fine. The important thing is that we know what the true mags are. One thing we did find is that the magnifications are consistantly slightly lower in value at 100kV than at 80kV. This surprised me at first but I am assuming that the difference is due to the higher acceleration of the electrons at 100kV. Am I correct to assume that higher energy electrons would be influenced less by the lenses, resulting in lower x-over point, which could result in a slight decrease in the magnification? I suspect that newer microscopes could compensate for this through the software programming and will be doing the calibration at different kV values on our CM-10 to verify this. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id JAA03101 for dist-Microscopy; Wed, 24 Jan 2001 09:21:00 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id JAA03098 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 24 Jan 2001 09:20:30 -0600 (CST) Received: from tumor.soft-imaging.com (tumor.soft-imaging.com [209.180.246.90]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id JAA03088 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 24 Jan 2001 09:20:18 -0600 (CST) Received: from ntserver3.soft-imaging.com (ntserver3 [192.168.5.2]) by tumor.soft-imaging.com (8.8.5/8.8.5) with ESMTP id IAA24150; Wed, 24 Jan 2001 08:13:35 -0600 Received: by sisus-hq-pdc with Internet Mail Service (5.0.1461.28) id {DF4V1FJ0} ; Wed, 24 Jan 2001 08:14:02 -0700 Message-ID: {D1B635C6092DD411B0700020781025DE0B69F6-at-sisus-hq-pdc} "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com}
Allen,
I am not sure I am following you. You say
"The best high energy experiments have been unable to assign a 'mass' to the electron."
To my knowledge and from a list of "fundamental constants" in my "Solid State Physics" book by Ashcroft and Mermin, the rest mass of an electron is 9.1095 x 10^(-31) kg. Just multiply that wit 1/2 v*v and you have the kinetic Energy of the electron. Of course you have to take into account relativistic effects. Or just multiply it with v and you have the momentum.
I think, you mistake the mass of the electron with that of the neutrino, which indeed they have not been able to determine, although there are indications from recent high energy physics measurements. But that's irrelevant here.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Allen R. Sampson [mailto:ars-at-sem.com] Sent: Monday, January 22, 2001 4:21 AM To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com'
Hi, all,
I'm back to ask for more help. Thanks in advance, before I forget to My colleague is gathering some information on Tektronix Phaser 950 color printers. Specifically she would like to know:
- the quality of b/w prints (are they publication quality?) Some color printers make wonderful color prints and awful b/w prints.
- what sort of paper we would need for the best b/w prints.
Any comments, pros and cons from users, or people who have tried this product would be very welcome.
Please contact me offline and I'll forward the replies to her.
Thanks again.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Yes, the focussing action of the lens, as given by d2z/dr2, is proportional to (field) squared over voltage. [Check, e.g. Joy, Romig & Goldstein, Principles of Analytical Electron Microscopy, pages 44-45.] I'l be interested to hear whether newer TEMs do compensate for this or not.
Gill ----- Original Message ----- } From: "Debby Sherman" {dsherman-at-purdue.edu} To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 24, 2001 7:45 AM
Hi all, I just wanted to run some interesting calibration data by you. We are working with some small particles and have to have reasonable size calibrations at both 80 and 100kV on a Philips EM-400 microscope (~20 years old). Calibration using replica gratings and catalase lattice spacing give us values somewhat below the magnifications indicated by the mag. readout on the panel. This is fine. The important thing is that we know what the true mags are. One thing we did find is that the magnifications are consistantly slightly lower in value at 100kV than at 80kV. This surprised me at first but I am assuming that the difference is due to the higher acceleration of the electrons at 100kV. Am I correct to assume that higher energy electrons would be influenced less by the lenses, resulting in lower x-over point, which could result in a slight decrease in the magnification? I suspect that newer microscopes could compensate for this through the software programming and will be doing the calibration at different kV values on our CM-10 to verify this. Debby
Dear Debby,
It is true that the lens currents will be higher at 100 kV than at 80 kV; however, even a 20-year-old microscope takes this into account and sets the lens currents to match the accelerating voltage. It seems that in your case this correction is slightly off resulting in the systematically lower mags. The newer microscopes will also set the lens currents to match the voltage, but whether this is done through software or electromechanically I don't know.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I have a small inner room within my office that is 6.2 5X 3.8 ft. with a high ceiling. The building was built about 1950 with some of the same electrical features. I would like to purchase a microtome (basic for cutting thin sections of plant tissue for TEM) for the little room, and the electrician asked if there were special requirements before the purchase is made. Could someone help me?
Thanks,
Nancy Robertson Research Plant Pathologist USDA/ARS 533 E. Fireweed Ave. Palmer AK 99645
I have been trying for a while to obtain EFTEM images on a series of polymers, but they wouldn't cooperate. Our group is investigating ionomers (i.e polymers with a hydrophobic backbone and approx. 5 mol % of either sulfonic acid or carboxylic acid groups that are neutralized with different metal ions). We know from STEM that these metal ion neutralized acid groups form aggregates in the 2 nm range within a hydrophobic matrix and we would like to know the elemental distribution of metal ions and O (and S in the case of SO3H). I use a JEOL210F FEG with GIF at 200kV. Has anybody ever tried to resolve such small structures by EFTEM ? I'd appreciate tips and tricks as well as some useful references.
Thanks for your help,
Andreas
************************************************* Dr. Andreas Taubert Materials Science and Engineering Dept. 3231 Walnut Street The University of Pennsylvania=20 Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
I've just run into a problem (and, I think, partially solved it) that may affect a lot of beam instruments with water-cooled diffusion pumps. We'd been having some problems lately with at least one of the diff pumps on our ESEM shutting itself off because it thought it wasn't being cooled enough. I checked our little Neslab chiller unit; it seemed OK, and since I'd just replaced the pump in it last year, I figured that there really couldn't be a problem with that.... So I was starting to worry that maybe we had a diff pump or at least a sensor starting to fail, but then this morning, the other diff pump also shut itself down. Hello, thinks I, then this has to be a cooling problem. So I disconnect the outflow from the chiller, and check the flow. Damn, lots of water coming out....must be something else. But then I get a bright idea....let's check the inline filter. Turns out, the filter is getting a bit full of crud. Green crud. Ok, so I put a new one in. Then I get a second bright idea (my personal daily limit). Let's check the water pressure/flow returning from the ESEM. ....Oh oh, very low. So I disconnect both inlet and outlet lines from the ESEM, stick the nozzle of a can of compressed air in one, and let fly. Wish I hadn't had the other line aimed at myself just then, but that's beside the point. About a half a litre of pretty dirty water came shooting out, including a surprising number of little solid bits of stuff. Judging by their blue-green colour, they're copper oxides, presumably from the water lines in my slowly-decomposing ESEM. Anyway, after that system flush, the flow seems a lot better, and now the diff pumps seem to be staying on. I don't believe it says anywhere in the manual that one should regularly do this kind of procedure on one's beam instrument, but I wonder if this is a common problem. Maybe some of you do a flush like this on a regular (annual?) basis? The water in our system is RO, and I don't put anything in to treat for algae, since that doesn't seem to be a problem. Yet there's obviously some corrosion going on. Is there a way to buffer the water to ensure a neutral pH that will certainly not bother the instrument? I'm assuming our water is a bit on the acid side, for this corrosion to be happening (?).
F. C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia Canada
Fellow colleagues, I am posting this for Carlos Saenz, a veteran histotec and licenced HT, in San Jose, Costa Rica, English is limited, but knowledge of histotecnology is excellent and commitment is there. He may be reached at 732-748-069 in New Jersey. I had one reply for Carlos to try and increase his English skills, and he is trying, but will someone give him an opportunity. I have been invited to lecture about Histotecnology in Mexico, Guatemala and Central America; And, the only countries I have not been in South America are Venezuela, Colombia, Guyanas, Brazil and Uruguay. And the only diffrence between the histotecnologist there and in the USA are opportunities and language barriers, othere than that, knowledge of our field is excellent. Thanks again, Teresa
Hi all till now answered people are right - the lens currents are adjusted according to selected HT by means of sets of analog resistors and reference voltages adjustable inside the EM400. If this adjustment is not 100% according to theory (what is allmost 100% true after so many years) than the image size projected on the screen differs from expected size and mag values can be different for each HT, non linear thru HT etc. It is a fine tuning required by service..... but is this mag accuracy worth this service work ?? maybe just do in series with shots with importance of mag value the one with same condition and calibration sample inserted for corrections as reference ??
----- Wiadomooæ oryginalna ----- Od: Debby Sherman {dsherman-at-purdue.edu} Do: message to: MSA list {microscopy-at-sparc5.microscopy.com} Wys³ano: 24 stycznia 2001 15:45 Temat: TEM calibration
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } I just wanted to run some interesting calibration data by you. We are working with some small particles and have to have reasonable size calibrations at both 80 and 100kV on a Philips EM-400 microscope (~20 years old). Calibration using replica gratings and catalase lattice spacing give us values somewhat below the magnifications indicated by the mag. readout on the panel. This is fine. The important thing is that we know what the true mags are. } One thing we did find is that the magnifications are consistantly slightly lower in value at 100kV than at 80kV. This surprised me at first but I am assuming that the difference is due to the higher acceleration of the electrons at 100kV. Am I correct to assume that higher energy electrons would be influenced less by the lenses, resulting in lower x-over point, which could result in a slight decrease in the magnification? } I suspect that newer microscopes could compensate for this through the software programming and will be doing the calibration at different kV values on our CM-10 to verify this. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907 }
I encountered a similar (green sludge and corrosion) problem on the DP's and recirculating water system of our TEM several years ago. I assumed that it was a reaction between the iron and copper tubing used in the system. Old time plumbers, for example, always warned about directly connecting iron and copper tubing because of the corrosion that would occur. I KNOW that on our DP's the iron fittings have copper connections. Oh well....
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
----- Wiadomooæ oryginalna ----- Od: Peggy Bisher {peggy-at-research.nj.nec.com} Do: Krzysztof Herman {kherman-at-labsoft.com.pl} Wys³ano: 24 stycznia 2001 21:44 Temat: Re: PD: TEM calibration
} } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } Hi } } all till now answered people are right - the lens currents are adjusted } } according to selected HT by means of sets of analog resistors and reference } } voltages adjustable inside the EM400. If this adjustment is not 100% } } according to theory (what is allmost 100% true after so many years) than the } } image size projected on the screen differs from expected size and mag values } } can be different for each HT, non linear thru HT etc. } } It is a fine tuning required by service..... but is this mag accuracy worth } } this service work ?? maybe just do in series with shots with importance of } } mag value the one with same condition and calibration sample inserted for } } corrections as reference ?? } } } } regards } } Krzysztof Herman } } } } EMISJA S.c. } } 02-892 Warszawa, ul.Bazancia 45 A, Poland } } tel/fax: (+48 22)6449753, 6449750 } } tel: (+48 601)307456 } } kherman-at-labsoft.com.pl } } www.emission.com.pl } } } I do remember at one time that Philips would guarantee that the } "nominal mag" be +/- 5% of the actual mag no matter what kV you were } using. What I am saying is that if you calibrate your microscope and } you found it to be more or less than 5% of what is actually displayed } on the panel then they should come in and make an adjustment for } you, providing you have a maintenance contract. } Many years ago I found one of my EM400's to be in some instances } 10-15% out so they fixed it. Otherwise it is your responsibility to } calibrate your microscope.
I am the service specialist grown from CM series onwards. I was maintening not only Philips microscopes. But each time I touch the EM-xxx machine I feel the smell of nobel time of Philips name. Now all this become a business, very commercial and without soul. Software makes it like this. It is similar difference like the old mechanical swiss clock 100 years old and modern Taiwan made electronic one. Do You feel the difference ? From otehr side the people from the times You mentioned (times of "customer first" slogan) "they should come and adjust it..." so "they" are allmost all retired or gone. I think - be happy that this EM-XXX still works. Ofcourse - pure customers point of view and feeling can differ from mine. All the best Krzysztof Herman EMISJA S.c. 02-892 Warszawa, ul.Bazancia 45 A, tel/fax: (+48 22)6449753, 6449750 tel: (+48 601)307456 kherman-at-labsoft.com.pl www.emission.com.pl
I suspect (but have not worked out), another possibility could be that the lens currents are set properly for 100kV but your HT tank is actually providing somewhat less..or more? Without thinking too hard, it seems like } 100kV could result in lower than expected mags. Is this error consistent over weeks, months? My EM300 calibrates consistently higher than published values, but I haven't seen a systematic difference with kV. In any case, if you have service on the instrument, ask the engineer to check currents, reference resistors, etc. The correct values should be easy to find.
Matt
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
} } } Dear Debby, } } It is true that the lens currents will be higher at 100 kV than at 80 } kV; } however, even a 20-year-old microscope takes this into account and sets } the lens currents to match the accelerating voltage. It seems that in } your case this correction is slightly off resulting in the } systematically } lower mags. The newer microscopes will also set the lens currents to } match the voltage, but whether this is done through software or } electromechanically I don't know. } } Yours, } } } Bill Tivol } Wadsworth Center } Albany NY } (518) 473-7399 WFT02-at-health.state.ny.us }
Is anyone using/used in the past Image Pro Express software (or Image Pro Plus)? I'm unfamiliar with the package, and would like to hear any comments (+ or -).
Thanks!
Tamara
(note new home bench!) |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
G'day Frank, Early last year we had a similar problem on our E3 ESEM, the water lines were blocking up with gelatinous material. After many attempts to reverse flush the lines we decided to pull all the water lines off and replace them. Our problem was in the water cooled alloy heatsink board, the alloy was slowly dissolving. We reamed out the alloy and fitted copper tube through the cooling channel. This solved the problem. Regards JVN
Frank Thomas wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers - } } I've just run into a problem (and, I think, partially solved it) that } may affect a lot of beam instruments with water-cooled diffusion pumps. } We'd been having some problems lately with at least one of the diff } pumps on our ESEM shutting itself off because it thought it wasn't being } cooled enough. I checked our little Neslab chiller unit; it seemed OK, and } since I'd just replaced the pump in it last year, I figured that there } really couldn't be a problem with that.... } So I was starting to worry that maybe we had a diff pump or at least a } sensor starting to fail, but then this morning, the other diff pump also } shut itself down. Hello, thinks I, then this has to be a cooling problem. So } I disconnect the outflow from the chiller, and check the flow. Damn, lots of } water coming out....must be something else. But then I get a bright } idea....let's check the inline filter. Turns out, the filter is getting a } bit full of crud. Green crud. Ok, so I put a new one in. Then I get a second } bright idea (my personal daily limit). Let's check the water pressure/flow } returning from the ESEM. ....Oh oh, very low. So I disconnect both inlet and } outlet lines from the ESEM, stick the nozzle of a can of compressed air in } one, and let fly. Wish I hadn't had the other line aimed at myself just } then, but that's beside the point. About a half a litre of pretty dirty } water came shooting out, including a surprising number of little solid bits } of stuff. Judging by their blue-green colour, they're copper oxides, } presumably from the water lines in my slowly-decomposing ESEM. } Anyway, after that system flush, the flow seems a lot better, and now } the diff pumps seem to be staying on. } I don't believe it says anywhere in the manual that one should regularly } do this kind of procedure on one's beam instrument, but I wonder if this is } a common problem. Maybe some of you do a flush like this on a regular } (annual?) basis? } The water in our system is RO, and I don't put anything in to treat for } algae, since that doesn't seem to be a problem. Yet there's obviously some } corrosion going on. Is there a way to buffer the water to ensure a neutral } pH that will certainly not bother the instrument? I'm assuming our water is } a bit on the acid side, for this corrosion to be happening (?). } } F. C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Dartmouth, Nova Scotia } Canada
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
A law of nature: Small diameter tubes in a heated system crud up. Filters must be cleaned and anti-corrosion fluids renewed. Check the flow rate (litres per minute) of the return line every couple of weeks. A thermometer measuring the temperature of the returned water is useful. Return lines from each instrument are best hooked into a 50mm gravity drain, which obviously must be higher than the cooling water reservoir. Such an open drain avoids back-pressure and makes monitoring of different instruments easier.
After some years EMs will need to be reversed flushed with water. If the flow is still not good enough, than pumping of a weak acid (10% acetic/vinegar) through the instrument in a loop may be required . There was a long discussion on anti corrosion items a couple of years ago. The Tips and Ticks site (one of the first on our Links page) may have that information more easily accessible than wading through the archives. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, January 25, 2001 5:51 AM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] wrote: } } Listers - } } I've just run into a problem (and, I think, partially solved it) that } may affect a lot of beam instruments with water-cooled diffusion pumps. } We'd been having some problems lately with at least one of the diff } pumps on our ESEM shutting itself off because it thought it wasn't being } cooled enough. I checked our little Neslab chiller unit; it seemed OK, and } since I'd just replaced the pump in it last year, I figured that there } really couldn't be a problem with that.... } So I was starting to worry that maybe we had a diff pump or at least a } sensor starting to fail, but then this morning, the other diff pump also } shut itself down. Hello, thinks I, then this has to be a cooling problem. So } I disconnect the outflow from the chiller, and check the flow. Damn, lots of } water coming out....must be something else. But then I get a bright } idea....let's check the inline filter. Turns out, the filter is getting a } bit full of crud. Green crud. Ok, so I put a new one in. Then I get a second } bright idea (my personal daily limit). Let's check the water pressure/flow } returning from the ESEM. ....Oh oh, very low. So I disconnect both inlet and } outlet lines from the ESEM, stick the nozzle of a can of compressed air in } one, and let fly. Wish I hadn't had the other line aimed at myself just } then, but that's beside the point. About a half a litre of pretty dirty } water came shooting out, including a surprising number of little solid bits } of stuff. Judging by their blue-green colour, they're copper oxides, } presumably from the water lines in my slowly-decomposing ESEM. } Anyway, after that system flush, the flow seems a lot better, and now } the diff pumps seem to be staying on. } I don't believe it says anywhere in the manual that one should regularly } do this kind of procedure on one's beam instrument, but I wonder if this is } a common problem. Maybe some of you do a flush like this on a regular } (annual?) basis? } The water in our system is RO, and I don't put anything in to treat for } algae, since that doesn't seem to be a problem. Yet there's obviously some } corrosion going on. Is there a way to buffer the water to ensure a neutral } pH that will certainly not bother the instrument? I'm assuming our water is } a bit on the acid side, for this corrosion to be happening (?). } } } F. C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Dartmouth, Nova Scotia } Canada }
I am assembling platform and poster presentations for a symposium on "Teaching Microscopy" to be given at the upcoming MSA meeting in Long Beach from August 5-9, 2001. I encourage anyone with an interest or experience with this topic to submit a presentation. Please contact me directly for more information or to answer any questions. See you in Long Beach!
Teaching Microscopy
Remote links to various light and electron microscopes, virtual microscopes, and CD-ROMS of images are all being used to teach colleagues and/or students about microscope theory, operation, and utilization. These are the new hardware and software tools for instruction. Are they effective? How are these tools being integrated into training programs and school curricula? What should be included in a training course? What is the best way to teach these concepts? This symposium will highlight examples of the hardware and software tools, as well as discuss different approaches, from formal classes and image collections to intensive workshops, to be used to teach microscopy theory and operation.
Steve Barlow
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
We use DI water and ethylene glycol or some good old fashioned anti-freeze. However, I can't remember the concentration right now. I think our "corrosion" problems promptly disappeared.
-----Original Message----- } From: Frank Thomas [mailto:thomasf-at-AGC.BIO.NS.CA] Sent: Wednesday, January 24, 2001 12:51 PM To: Microscopy-at-sparc5.microscopy.com
Listers -
I've just run into a problem (and, I think, partially solved it) that may affect a lot of beam instruments with water-cooled diffusion pumps. We'd been having some problems lately with at least one of the diff pumps on our ESEM shutting itself off because it thought it wasn't being cooled enough. I checked our little Neslab chiller unit; it seemed OK, and since I'd just replaced the pump in it last year, I figured that there really couldn't be a problem with that.... So I was starting to worry that maybe we had a diff pump or at least a sensor starting to fail, but then this morning, the other diff pump also shut itself down. Hello, thinks I, then this has to be a cooling problem. So I disconnect the outflow from the chiller, and check the flow. Damn, lots of water coming out....must be something else. But then I get a bright idea....let's check the inline filter. Turns out, the filter is getting a bit full of crud. Green crud. Ok, so I put a new one in. Then I get a second bright idea (my personal daily limit). Let's check the water pressure/flow returning from the ESEM. ....Oh oh, very low. So I disconnect both inlet and outlet lines from the ESEM, stick the nozzle of a can of compressed air in one, and let fly. Wish I hadn't had the other line aimed at myself just then, but that's beside the point. About a half a litre of pretty dirty water came shooting out, including a surprising number of little solid bits of stuff. Judging by their blue-green colour, they're copper oxides, presumably from the water lines in my slowly-decomposing ESEM. Anyway, after that system flush, the flow seems a lot better, and now the diff pumps seem to be staying on. I don't believe it says anywhere in the manual that one should regularly do this kind of procedure on one's beam instrument, but I wonder if this is a common problem. Maybe some of you do a flush like this on a regular (annual?) basis? The water in our system is RO, and I don't put anything in to treat for algae, since that doesn't seem to be a problem. Yet there's obviously some corrosion going on. Is there a way to buffer the water to ensure a neutral pH that will certainly not bother the instrument? I'm assuming our water is a bit on the acid side, for this corrosion to be happening (?).
F. C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia Canada
I used to put antifreeze in the chillers but in the later models there is a warning stating not to use antifreeze as it damages the seals.
I would like to know what everyone now uses to prevent organic growth.
By the way the DP's must have been fairly warm to trip the sensors. If I have any questions at all about the temperture Is usually feel the DP cooling coils. All the coils should be cool except for the last two or three turns. Typical temperature is 20 deg C.
Earl Weltmer ----- Original Message ----- } From: "Frank Thomas" {thomasf-at-AGC.BIO.NS.CA} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 24, 2001 11:51 AM
Earl said: I used to put antifreeze in the chillers but in the later models there is a warning stating not to use antifreeze as it damages the seals.
I would like to know what everyone now uses to prevent organic growth.
We dont put anything in the water these days, but check filters, flush and reverse flush once a year or whenever convenient or when the temperature or flow meters look ominous. Our recirculating system has a 500 litre buffer tank which is cleaned about every five years.
We have had certainly no more trouble, probably less, than when we put various chemicals into the water
Sally
Dr Sally Stowe Facility Coordinator, ANU EMU Box 475 Canberra ACT 2601 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 or 6125 8525 http://www.anu.edu.au/EMU
Dear Listers, I'm trying to obtain 8 um thick sections of non-decalcified bone which has been embedded in methacrylate using a R-J autocut. Thanks in advance for any tips to minimize compression and adhere sections to a glass slide. Rosemary Walsh EMF for the Life Sciences Penn State University
all ultramicrotomes I know run either with a standard electrical supply (eg 110 V in the USA, 220 V in many other countries eg here in Austria) or with a "full range power supply". Our Leica EM UCT and UCR are equipped with a full range power supply for a voltage input range from "90 - 260 VAC, 50-60 Hz" (as the engineers call it). Some older model you will have to switch between voltages depending on the voltage you use...
You may want to run additionally a few desktop lights, a LM for looking at semis before trimming down to a blockface for ultrathins, etc. but this should be no problem even with electrical features from the 1950`s.
Important when deciding for a UM room: Are you having vibrations in this room? In which floor is your room? Are you having a concrete floor? and what is the floor covering made of? These factors all may affect the stability of your room!
I strongly recommend the use of an antivibration instrument table or AT LEAST an antivibration base plate for the UM. Do not use a desk or smthg similar! You can test for vibrations easily by putting a petridish filled with water (slightly "overfilled" with a convex meniscus) on a table in your room under a light - you should see NO waves! Slam the door and watch what happens :-)
And just one more thought REALLY important: Should you ever decide to work with a cryosectioning equipment: DANGER OF SUFFOCATION!!! Make sure that you have a good ventilation of your room! 1 litre of liquid nitrogen (LN2) will produce 700 litres of gaseous nitrogen (GN2). It is odourless and tasteless and an operator in a small room may suffocate and not even recognize that he/she is in danger!!! You may use an oxygen analyzer (range 0 - 25%) for testing ...
Hope that helps you,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Maybe US plumbers are better trained, but UK heating systems are almost always iron (pressed steel steel) radiators with copper connecting pipes. Other metals involved are brass valves and connectors, and lead/tin solder at soldered joints. Systems like that corrode unless inhibitors are used. The sludge that collects in my copper/iron central heating system is deep black, like indian ink, not green. Is it possible that your green sludge is not copper but algae?? Transparent plastic tubing in the system may allow alagal growth, provided there is some source of carbon. Bacterial/fungal decomposition of glycol could provide that. Chris
----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 24, 2001 9:03 PM
Debby
there are several things that you should consider when checking magnification on a TEM: First of all most manufacturers will aim for a consistency of about +/-5% accuracy because of the laws of diminishing returns. Also they may have an optimum voltage Second there will be variations up and down the magnification scale because a balance has been struck between several lenses and when they are excited differently this will be expressed differently. Third remember that there is a very short working distance between lens polepiece and specimen (just a few millimetres) so if the specimen height varies by +/- 0.2 mm this will greatly affect the magnification. For instance if the grid is slightly bent or kinked, if you don't check magnifications at a standard Z position (eucentric if that's available) then you will get inconsistencies of easily 5%. When I calibrated at eucentric I noticed that all of my magnifications at 75kv were near or better than +/- 4% whereas one or two previous sets were mostly worse than +/- 5% Fourth all of the above will change with time as the electronics changes so calibrations, if you're fussy, should be done perhaps every 6 months or year if you're fussy or notice great changes. I'm sure that Steve Chapman would agree that it's an excellent way of monitoring part of the performance of your microscope anyway. Fifth consider how you measure your calibrations you should always measure the negative never the print (use a calibrated graticule eyepiece or travelling microscope). Prints can vary by 1 or 2% so easily. Sixth the calibration standards for TEM will be at best +/- 2% accurate unless you've got a certificate that says otherwise. They will often differ e.g. catalase and diffraction grating will give you a slightly different answer. Seventh don't forget the hysteresis factor. On most microscopes you get best calibration by calibrating at the highest magnification then working down the range because of inherent hysteresis in the electromagnetic lenses. Some manufacturers even used to supply a special button to overcome this on our old AEI 801 it was called standardise magnification. Eighth I'm slipping I can usually come up with about ten problems.
Never quote magnifications to 3 significant figures it's meaningless. It may sound insurmountable but if you are aware of the problems you can reduce or avoid them. If you have calibrated and see little change between sets then you can put confidence limits on your accuracy. If it's critical that a user needs accuracy then photograph a consecutive shot of a calibration sample under exactly the same conditions.
good luck in your quest for the perfect magnification.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland
Debby Sherman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all, } I just wanted to run some interesting calibration data by you. We are working with some small particles and have to have reasonable size calibrations at both 80 and 100kV on a Philips EM-400 microscope (~20 years old). Calibration using replica gratings and catalase lattice spacing give us values somewhat below the magnifications indicated by the mag. readout on the panel. This is fine. The important thing is that we know what the true mags are. } One thing we did find is that the magnifications are consistantly slightly lower in value at 100kV than at 80kV. This surprised me at first but I am assuming that the difference is due to the higher acceleration of the electrons at 100kV. Am I correct to assume that higher energy electrons would be influenced less by the lenses, resulting in lower x-over point, which could result in a slight decrease in the magnification? } I suspect that newer microscopes could compensate for this through the software programming and will be doing the calibration at different kV values on our CM-10 to verify this. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907
There is no need to scrape off the old wax when reusing Trufs - just put them on the multiplate and it melts and gets recycled. I used to to re-use them then I hit a problem where I had difficulty wetting the knife edge. I got so desperate I decided to use a new boat each time.
Dave
On Wed, 24 Jan 2001 12:51:47 +0100 "joachim.prutsch-at-leica-microsystems.com"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi list, } } may I just add to the discussion about the knife boats: } } yes, if you use dental wax to glue the Truf to the knife (with the Leica } Multiplate, especially designed for that purpose its really easy) the boat } can be reused a few times. } } HOWEVER: } 1. breaking off the Truf from the knife and scratching away the old } hardened wax may damage the Truf - leaking may result from this! Can be } frustrating if you have perfect sections of a precious sample and see them } dive away ... } 2. Esp. with ultrathins (but also with semithins) you must be aware of } possible cross contamination of old sections to the new set (I have no idea } if there is a GLP rule on that but in a hospital mixing up different } samples might cause serious problems)! } } BTW the "LKB"Trufs are "Leica"Trufs now, but call them as you like it :-) } } Best regards, } } Joachim } } Dr. Joachim Prutsch } Product Manager EM Specimen Preparation } } Leica Microsystems GmbH } Hernalser Hauptstr. 219 email: } Joachim.Prutsch-at-leica-microsystems.com } A 1170 Vienna Tel.: +43 1 4 88 99 - 235 } AUSTRIA Fax: +43 1 4 88 99 - 350 } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
One point about magnification calibration that I have not read here yet is that for a particular nominal setting, the actual mag differs when the mag has been stepped up compared to stepping down the mag. I have been told this is due to hysteresis in the lenses (predominantly the objective probably). I have measured up to 10% difference.
I have not kept a close eye on this discussion, but the dependence of the electron voltage was mentioned. If this is so, that means that the electron accelerating voltage would need to be checked. I remember in the book by Forwood and Clarebrough (Electron Microscopy of Interfaces in Metals and Alloys, published by Adam Hilger) where they found the accelerating voltage of their instrument was 220 keV as opposed to 200 keV. I take it that could make a difference.
-- Dr. Steven Celotto Department of Engineering Materials Science & Engineering University of Liverpool Brownlow Hill Liverpool L69 3BX UNITED KINGDOM
At 5:16 PM -0800 1/24/01, Earl Weltmer wrote: } I would like to know what everyone now uses to prevent organic growth.
We sprinkle dichlorophene on the surface of the water in our Haskris chiller to prevent fungal/bacterial growth, and buffer the water to about pH 7.0 with sodium bicarbonate. So far (for the last 21 years), these procedures have worked fine.
I used the working demo version of Image Pro Plus during the summer to help analyze my SEM photos for my masters thesis. I found the program to be very easy to use and very functional. I was really pleased with how well you could set, save, and recall image calibrations for doing measurement work. I had SEM micrographs from two different machines (one acquired digitally, the other Polaroid film) and each had a different screen calibration.
dz
-----Original Message----- } From: tamara a howard [mailto:thoward-at-UNM.EDU] Sent: Wednesday, January 24, 2001 6:16 PM To: Microscopy-at-sparc5.microscopy.com; histonet-at-pathology.swmed.edu
Is anyone using/used in the past Image Pro Express software (or Image Pro Plus)? I'm unfamiliar with the package, and would like to hear any comments (+ or -).
Thanks!
Tamara
(note new home bench!) |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
I heard the same about the antifreeze...checked with Haskris and they recommended (a year or two ago) Dichlorophene to prevent biological growth. Seems to work but I still need to clean filters 2-3 times a year and monitor flow. Haskris actually faxed me an info sheet, there's a chance I could find it so email me if you would like it.
Matt
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Wednesday, January 24, 2001 8:16 PM, Earl Weltmer [SMTP:eweltmer-at-home.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good question. } } I used to put antifreeze in the chillers but in the later models there is a } warning stating not to use antifreeze as it damages the seals. } } I would like to know what everyone now uses to prevent organic growth. } } By the way the DP's must have been fairly warm to trip the sensors. } If I have any questions at all about the temperture Is usually feel the DP } cooling coils. } All the coils should be cool except for the last two or three turns. Typical } temperature is 20 deg C. } } Earl Weltmer } ----- Original Message ----- } } From: "Frank Thomas" {thomasf-at-AGC.BIO.NS.CA} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, January 24, 2001 11:51 AM } Subject: Fun with vacuum systems } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Listers - } } } } I've just run into a problem (and, I think, partially solved it) that } } may affect a lot of beam instruments with water-cooled diffusion pumps. } } We'd been having some problems lately with at least one of the diff } } pumps on our ESEM shutting itself off because it thought it wasn't being } } cooled enough. I checked our little Neslab chiller unit; it seemed OK, and } } since I'd just replaced the pump in it last year, I figured that there } } really couldn't be a problem with that.... } } So I was starting to worry that maybe we had a diff pump or at least a } } sensor starting to fail, but then this morning, the other diff pump also } } shut itself down. Hello, thinks I, then this has to be a cooling problem. } So } } I disconnect the outflow from the chiller, and check the flow. Damn, lots } of } } water coming out....must be something else. But then I get a bright } } idea....let's check the inline filter. Turns out, the filter is getting a } } bit full of crud. Green crud. Ok, so I put a new one in. Then I get a } second } } bright idea (my personal daily limit). Let's check the water pressure/flow } } returning from the ESEM. ....Oh oh, very low. So I disconnect both inlet } and } } outlet lines from the ESEM, stick the nozzle of a can of compressed air in } } one, and let fly. Wish I hadn't had the other line aimed at myself just } } then, but that's beside the point. About a half a litre of pretty dirty } } water came shooting out, including a surprising number of little solid } bits } } of stuff. Judging by their blue-green colour, they're copper oxides, } } presumably from the water lines in my slowly-decomposing ESEM. } } Anyway, after that system flush, the flow seems a lot better, and now } } the diff pumps seem to be staying on. } } I don't believe it says anywhere in the manual that one should } regularly } } do this kind of procedure on one's beam instrument, but I wonder if this } is } } a common problem. Maybe some of you do a flush like this on a regular } } (annual?) basis? } } The water in our system is RO, and I don't put anything in to treat } for } } algae, since that doesn't seem to be a problem. Yet there's obviously some } } corrosion going on. Is there a way to buffer the water to ensure a neutral } } pH that will certainly not bother the instrument? I'm assuming our water } is } } a bit on the acid side, for this corrosion to be happening (?). } } } } } } F. C. Thomas } } MicroAnalysis Facility } } Geological Survey of Canada (Atlantic) } } Dartmouth, Nova Scotia } } Canada } } } }
Dear listers, Our 10 year old Ilford print processor is breaking down piece by piece. We've about reached the point of diminishing returns and so are considering replacing it. Not everyone has switched to digital so we use quite a lot of film. Can any of you recommend a good black and white print processor?
Thank you,
Mary Gail Engle Manager Electron Microscopy & Imaging Facility University of Kentucky
We need to determine a reasonable salary (at a research university) for someone with the following skills:
PhD degree in biological sciences 11 yr experience (practial & theory) in LM, SEM, TEM microscope maintenance (basic and advanced-mechanical) basic and advanced scope operator (STEM, EDS) excellent training skills (personable) excellent communication skills (verbal, written) specimen prep, ultramicrotomy, immunocytochemistry, vacuum techniques, digital imaging, statistics 1 yr exerience in atomic force microscopy 8 yr supervisory experience
This is a 12 month, continuing, non-tenured, hard-money position. The person provides research support to faculty & researchers on campus.
What are other academic institutuions paying such (or similar) individuals?
Thank you. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ through the North Carolina State University Department of Continuing and Professional Education is now in its 19th year. The course will be presented May 9 - 11, 2001, in Raleigh, NC, and June 6-8, 2001, at the Danish Technological Institute in Taastrup, Denmark (near Copenhagen). This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to
Hi all, Is it possible to fix plant tissue and have the stoma (guard cells and the pore between them) remain in their original position (opened or closed)? Does anyone have a reference? thanks for the help, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Hello John Surely you will hear of people making less or more but between the survey done by Don Grimes of Microscopy Today (457 microscopist) & the salary survey of optically educated individuals published annually in Photonics Spectra, I think your window is 50-70K$. Can you get someone for less, probably. Would you like continuity in the years to come? (the answer is yes) One more thing, I don't think the range of skills & experience you are asking for are typical of microscopist in education e.g. ignore the educational discount usually applied to most of us.
Bruce Brinson Rice U.
"John J. Bozzola" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We need to determine a reasonable salary (at a research university) } for someone with the following skills: } } PhD degree in biological sciences } 11 yr experience (practial & theory) in LM, SEM, TEM } microscope maintenance (basic and advanced-mechanical) } basic and advanced scope operator (STEM, EDS) } excellent training skills (personable) } excellent communication skills (verbal, written) } specimen prep, ultramicrotomy, immunocytochemistry, vacuum } techniques, digital imaging, statistics } 1 yr exerience in atomic force microscopy } 8 yr supervisory experience } } This is a 12 month, continuing, non-tenured, hard-money position. } The person provides research support to faculty & researchers on campus. } } What are other academic institutuions paying such (or similar) individuals? } } Thank you. } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ##############################################################
I'm sure there are more microscopy CD-ROMS out there than I know about. A very nice one that comes to mind is one produced by Oxford Instruments as part of its X-Ray microanalysis tutorial. That one concentrates mainly on SEM and microanalysis. It was on desplay at recent MSA meetings. Instructors are best served by going to the Project Micro website and checking that bibliography, so ably maintained by the tireless Caroline Schooley. If anyone knows about microscopy materials not on that list, please send me and Caroline the title and source (even better a review of the material and what level it is appropriate for).
While the major emphasis of Project Micro is for pre-college, Caroline has tried to include useful materials for all levels.
Steve
} From: "Barry Searle" {B.Searle-at-unsw.edu.au} } To: "Steve Barlow" {sbarlow-at-sunstroke.sdsu.edu} } Subject: Re: teaching microscopy } Date: Thu, 25 Jan 2001 12:13:46 +1100 } MIME-Version: 1.0 } X-Priority: 3} Steve, } } Would you have a web address for CD-ROMS on electron Microscopy and } teaching? } } Thanks } } Barry } EMU } UNSW } } } ----- Original Message ----- } From: Steve Barlow {sbarlow-at-sunstroke.sdsu.edu} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, January 25, 2001 11:51 AM } Subject: teaching microscopy } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I am assembling platform and poster presentations for a symposium on } } "Teaching Microscopy" to be given at the upcoming MSA meeting in Long } Beach } } from August 5-9, 2001. I encourage anyone with an interest or experience } } with this topic to submit a presentation. Please contact me directly for } } more information or to answer any questions. See you in Long Beach! } } } } Teaching Microscopy } } } } Remote links to various light and electron microscopes, virtual } } microscopes, and CD-ROMS of images are all being used to teach colleagues } } and/or students about microscope theory, operation, and utilization. These } } are the new hardware and software tools for instruction. Are they } } effective? How are these tools being integrated into training programs and } } school curricula? What should be included in a training course? What is } the } } best way to teach these concepts? This symposium will highlight examples } of } } the hardware and software tools, as well as discuss different approaches, } } from formal classes and image collections to intensive workshops, to be } } used to teach microscopy theory and operation. } } } } Steve Barlow } } } } } } } } ___________________________________________________ } } Dr. Steven Barlow } } EM Facility/Biology Dept. } } San Diego State University } } 5500 Campanile Drive } } San Diego CA 92182-4614 } } phone: (619) 594-4523 } } fax: (619) 594-5676 } } } } email: sbarlow-at-sunstroke.sdsu.edu } } http://www.sci.sdsu.edu/emfacility } } } } Chairman, Educational Outreach subcommittee } } promoting microscopy instruction and increased access to microscopes } } Microscopy Society of America } } http://www.msa.microscopy.com/ } } } } } } } } } }
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
We have an ETEC Autoscan whose main use is as a teaching scope and for doing demos for the various groups that tour our facility. It has TV capability which is really helpful for the above mentioned activities, but is also nice when you're searching for small particulates scattered all over the plug. Our TV Scan module is currently in for repair so we are without this useful accessory (and the class has started and the tours keep coming). We have two other ETECs that we use as spare parts, but neither of them had TV capability. Is there anybody out there having one of these (working) modules that would be willing to part with it? We might even be able to work out a trade if I have a module that you need. TIA. Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
I have been following a protocol that involves using tannic acid (1% in 50mM Na cacodylate buffer) to prevent shrinkage of cells in Xenopus. I am looking at cilia on the surface of the embryos and when I incubate in tannic acid I see a deposition of crystals on the surface of the embryos. I have made sure that the embryos as well as the solution is at room temperature before and during incubation but this problem persists. I am also washing subsequently in cacodylate buffer three times. Has anyone seen this before or have any suggestions about preventing it? The tannic acid does appear to reduce cell shrinkage but makes it difficult to observe the cilia. Julaine Roffers University of Wisconsin-Milwaukee
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I used to use Image Pro for image analysis and diffraction pattern measurements. It is pretty good. Sometime you have to make calibration in order to measure diffraction patterns. i hope there will be a version for Mac. ( i am now using NIH on Mac)
Yali Tang
Materials Science Division Argonne National Laboratory Bldg. 223, A109 Argonne, IL 60439 Ph: 630-252-6219 Fax:630-252-7777 Email:ytang-at-anl.gov
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Listmembers,
For those interested in what Haskris recommended to me: I scanned in their sheet and OCR'd it so I might have missed some "typos". Other standard disclaimers: this info is two years old so you may want to check again with Haskris, or whichever manufacturer applies in your case.
Matt
WATER TREATMENT FOR BIOLOGICAL GROWTH In reference to our recent phone conversation, following are some suggestions regarding water treatment for biological growth. Please keep in mind that any additive must have the prior approval of the manufacturer of the equipment being cooled.
It is important that you fill the system with clean, potable distilled water.
Check the system after one week of operation, if algae is starting to form, flush the system in accordance with one of the following alternatives:
ALTERNATIVE 1: Add one cup of chlorine bleach per 15 gallons of water, circulate 20-30 minutes. Drain the system and refill with clean water.
ATLERNATIVE 2: Add one pint of hydrogen peroxide per 15 gallons of water. Circulate 20-30 minutes. Drain the system and refill with clean water.
Once the system has been flushed we recommend that you proceed with one of the following water additives:
ALTERNATIVE 1: A 10% solution of lab grade (no additives) ethylene glycol is recommended.
ALTERNATIVE 2: Dichlorophene. This is a non-soluble white powder. It is a fungicide/bactericide and should be sprinkled evenly across the entire tank's water surface.
Haskris Co. does not sell this product, but we have the following source for you to call:
ICN Biomedicals, Inc,. P.O. Box 5023, Costa Mesa, CA 92626 Phone: 800-854-0530
Dichlorophene Catalog # 05201981
ALTERNATIVE 3: Chlorine, 8 parts per million.
NOTE: A 5 micron filter is very helpful in keeping the system clean, especially if used in conjunction with one of the additives described above.
I have a researcher who needs an unusual type of gold-conj. secondary antibody. Her primary is made in goat and the only known available anti-goat is made in rabbit. Her tissue has cross-reactivity with rabbit. Is there any other species (donkey, dog, Brazilian opossum...) that anyone knows of as an anti-goat gold conjugate secondary? Do we have to make a home-made version? Thanks!
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
There is also a very good set of teaching tools for SEM, ESEM, and SEM/EDS done by Brendan Griffin (UWestern Australia - Perth), Clive Nockolds (UofSydney) it is University level. Contact Brendan for details at {bjg-at-cyllene.uwa.edu.au}
Nestor Your Friendly Neighborhood SysOp
=========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
Does anyone have a manual for a AO 923 knife sharpener that they would be willing to copy and mail or fax to me? This is the model that automatically sets the plate heights for the coarse and fine blade facets.
Thank you,
Jaclynn Lett, Research Assistant jlett-at-cid.wustl.edu
Central Institute for the Deaf Faye and Carl Simons Center for Biology of Hearing and Deafness 4560 Clayton Ave. St. Louis, MO 63110
Can anyone help this person . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Question: I have a Zeiss Jena microscope drawing set (Nr.6454) Stamped Zeichenapparat on the box. I am looking for directions for this particular instrument or a similar type.
Fellow Microscopists: As some one on the metallurgical side of things, I may be able to help. Chris Jeffry is on the right track. The corrosion product from Cu/Fe couples. such Cu pipe and Fe fittings, may be magnetite, a black, magnetic residue or, maybe, red rust but not green Cu compounds. In such a couple, Fe corrodes and the Cu does not. I'd bet on algae as the green material. You biologists ought to be able to tell if it's algae.
regards,
Sam Purdy Technical Center National Steel Corp.
} ---------- } From: Chris Jeffree } Sent: Thursday, January 2001, 4:49 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Fun with vacuum systems } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Maybe US plumbers are better trained, but UK heating systems are } almost always iron (pressed steel steel) radiators with copper } connecting pipes. Other metals involved are brass valves and } connectors, and lead/tin solder at soldered joints. Systems like that } corrode unless inhibitors are used. The sludge that collects in my } copper/iron central heating system is deep black, like indian ink, not } green. } Is it possible that your green sludge is not copper but algae?? } Transparent plastic tubing in the system may allow alagal growth, } provided there is some source of carbon. Bacterial/fungal } decomposition of glycol could provide that. } Chris } } ----- Original Message ----- } } From: "John J. Bozzola" {bozzola-at-siu.edu} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, January 24, 2001 9:03 PM } Subject: Re: Fun with vacuum systems } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } Hey, deja-vu all over again...... } } } } I encountered a similar (green sludge and corrosion) problem on the } } DP's and recirculating water system of our TEM several years ago. I } } assumed that it was a reaction between the iron and copper tubing } } used in the system. Old time plumbers, for example, always warned } } about directly connecting iron and copper tubing because of the } } corrosion that would occur. I KNOW that on our DP's the iron } fittings } } have copper connections. Oh well.... } } } } JB } } } } -- } } ############################################################## } } John J. Bozzola, Ph.D., Director } } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } } 750 Communications Drive - MC 4402 } } Southern Illinois University } } Carbondale, IL 62901 U.S.A. } } Phone: 618-453-3730 } } Fax: 618-453-2665 } } Email: bozzola-at-siu.edu } } Web: http://www.siu.edu/departments/shops/cem.html } } ############################################################## } } } }
Hi All Is there anybody out there who may have a spare Critical Point Drier to sell. We urgently need one, and our existing budget does not allow the purchase of a new one right now. Any offers???? We're hoping that someone in South Africa would respond to this offer. Makes life a bit easier. Thanks
Hi Malcolm, just thought I'd raise a few points. I should say I'm using a 20 year old JEOL 120CX and things may be different for more modern instruments!
1) I find that magnification is changed only slightly by variations in sample position (maybe 3% top to bottom of my 1 mm z-range). However diffraction pattern magnification (camera length) is changed enormously (+/- 50%). It's easy to see why if you draw a ray diagram. 2) I agree the standards are no better than +/- 2%. I'm lucky to be working in a field where I can generate my own standards which are an order of magnitude better (III-V superlattices which can be measured by high resolution X-ray diffraction very accurately). 3) If I turn all the knobs to the end and back I find magnification reproducibility is better than I can measure (about 0.5%). Unless, of course the reference batteries start going flat, but then the position of the knobs in the standard setting changes. I am happy to quote mags to 3 significant figures (but not 4!).
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Debby } } there are several things that you should consider when checking magnification on a TEM: } First of all most manufacturers will aim for a consistency of about +/-5% accuracy because of the laws of diminishing returns. Also they may have an optimum voltage } Second there will be variations up and down the magnification scale because a balance has been struck between several lenses and when they are excited differently this will be expressed differently. } Third remember that there is a very short working distance between lens polepiece and specimen (just a few millimetres) so if the specimen height varies by +/- 0.2 mm this will greatly affect the magnification. For instance if the grid is slightly bent or kinked, if you don't check magnifications at a standard Z position (eucentric if that's available) then you will get inconsistencies of easily 5%. When I calibrated at eucentric I noticed that all of my } magnifications at 75kv were near or better than +/- 4% whereas one or two previous sets were mostly worse than +/- 5% } Fourth all of the above will change with time as the electronics changes so calibrations, if you're fussy, should be done perhaps every 6 months or year if you're fussy or notice great changes. I'm sure that Steve Chapman would agree that it's an excellent way of monitoring part of the performance of your microscope anyway. } Fifth consider how you measure your calibrations you should always measure the negative never the print (use a calibrated graticule eyepiece or travelling microscope). Prints can vary by 1 or 2% so easily. } Sixth the calibration standards for TEM will be at best +/- 2% accurate unless you've got a certificate that says otherwise. They will often differ e.g. catalase and diffraction grating will give you a slightly different answer. } Seventh don't forget the hysteresis factor. On most microscopes you get best calibration by calibrating at the highest magnification then working down the range because of inherent hysteresis in the electromagnetic lenses. Some manufacturers even used to supply a special button to overcome this on our old AEI 801 it was called standardise magnification. } Eighth I'm slipping I can usually come up with about ten problems. } } Never quote magnifications to 3 significant figures it's meaningless. It may sound insurmountable but if you are aware of the problems you can reduce or avoid them. If you have calibrated and see little change between sets then you can put confidence limits on your accuracy. If it's critical that a user needs accuracy then photograph a consecutive shot of a calibration sample under exactly the same conditions. } } good luck in your quest for the perfect magnification. } } Malcolm } } Malcolm Haswell } e.m. unit } University of Sunderland } } } Debby Sherman wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi all, } } I just wanted to run some interesting calibration data by you. We are working with some small particles and have to have reasonable size calibrations at both 80 and 100kV on a Philips EM-400 microscope (~20 years old). Calibration using replica gratings and catalase lattice spacing give us values somewhat below the magnifications indicated by the mag. readout on the panel. This is fine. The important thing is that we know what the true mags are. } } One thing we did find is that the magnifications are consistantly slightly lower in value at 100kV than at 80kV. This surprised me at first but I am assuming that the difference is due to the higher acceleration of the electrons at 100kV. Am I correct to assume that higher energy electrons would be influenced less by the lenses, resulting in lower x-over point, which could result in a slight decrease in the magnification? } } I suspect that newer microscopes could compensate for this through the software programming and will be doing the calibration at different kV values on our CM-10 to verify this. } } Debby } } } } Debby Sherman Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-099e Whistler Building } } West Lafayette, IN 47907 }
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
I wrote a throw away comment at the end of my last mailing to the list about NEVER quoting magnifications to 3 significant figures. I think I should qualify that because 3 significant figures at 10k is different from 3 significant figures at 98k. It might be reasonable to quote 10.5k (this would depend on what % error your calibration gave) but pointless to quote 98.5k.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } Debby } } there are several things that you should consider when checking magnification on a TEM:
} {SNIP} } } Never quote magnifications to 3 significant figures it's meaningless. It may sound insurmountable but if you are aware of the problems you can reduce or avoid them. If you have calibrated and see little change between sets then you can put confidence limits on your accuracy. If it's critical that a user needs accuracy then photograph a consecutive shot of a calibration sample under exactly the same conditions. } } good luck in your quest for the perfect magnification. } } Malcolm } } Malcolm Haswell } e.m. unit } University of Sunderland } } Debby Sherman wrote: } } } ------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------.
} } } } Hi all, } } I just wanted to run some interesting calibration data by you. We are working with some small particles and have to have reasonable size calibrations at both 80 and 100kV on a Philips EM-400 microscope (~20 years old). Calibration using replica gratings and catalase lattice spacing give us values somewhat below the magnifications indicated by the mag. readout on the panel. This is fine. The important thing is that we know what the true mags are. } } One thing we did find is that the magnifications are consistantly slightly lower in value at 100kV than at 80kV. This surprised me at first but I am assuming that the difference is due to the higher acceleration of the electrons at 100kV. Am I correct to assume that higher energy electrons would be influenced less by the lenses, resulting in lower x-over point, which could result in a slight decrease in the magnification? } } I suspect that newer microscopes could compensate for this through the software programming and will be doing the calibration at different kV values on our CM-10 to verify this. } } Debby } } } } Debby Sherman Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-099e Whistler Building } } West Lafayette, IN 47907
Julaine: A question: how are you using the tannic acid? Is this in one of the OTOTO methods, or just part of the glut mixture? An answer to these would allow me to give you some advice/experience, as I have used both, but they are really different issues. I'll check back later today, and try to help then.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Thu, 25 Jan 2001 16:29:05 -0600, Julaine Roffers wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have been following a protocol that involves using tannic acid (1% in } 50mM Na cacodylate buffer) to prevent shrinkage of cells in Xenopus. I } am looking at cilia on the surface of the embryos and when I incubate in } tannic acid I see a deposition of crystals on the surface of the } embryos. I have made sure that the embryos as well as the solution is } at room temperature before and during incubation but this problem } persists. I am also washing subsequently in cacodylate buffer three } times. Has anyone seen this before or have any suggestions about } preventing it? The tannic acid does appear to reduce cell shrinkage but } makes it difficult to observe the cilia. } Julaine Roffers } University of Wisconsin-Milwaukee }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I used nine ultramicrotomes (including a cryo) over the years. All except for one Reichert, which had its own desk, were placed on a common office desk and we never had a hint of vibration problems. (Who remembers the hand-driven Porter-Blum, the Siroflex and the Huxley, great stuff - 35 years ago) However, my labs were either on the ground floor or in a basement. I suggest that you buy an expensive table only when needed.
Suffocation: sure its possible, but why scare the population. Our body does not require the nominal 16% of O2. The partial pressure of gases dictates that only about half the nominal is available at 10,000 ft altitude, which is equivalent to the cabin pressure of aircraft at high altitude. A small microtome room of 4x3x2.5m contains 30,000 liters of air and over 80% of that is nitrogen. I don't know how nitrogen hungry your equipment is, but I suggest that leaving the door open is good advice. If you have air-conditioning with return vents, you can even shut that door. I rather have a Canary than an oxygen analyser in the cryolab. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, January 25, 2001 4:44 PM, "joachim.prutsch-at-leica-microsystems.com" -at-sparc5.microscopy.com [SMTP:"joachim.prutsch-at-leica-microsystems.com"-at-sparc5.microscopy.com] wrote: } } Hi Nancy, }
snip} } Important when deciding for a UM room: Are you having vibrations in this } room? In which floor is your room? Are you having a concrete floor? and } what is the floor covering made of? These factors all may affect the } stability of your room! } } I strongly recommend the use of an antivibration instrument table or AT } LEAST an antivibration base plate for the UM. Do not use a desk or smthg } similar! You can test for vibrations easily by putting a petridish filled } with water (slightly "overfilled" with a convex meniscus) on a table in } your room under a light - you should see NO waves! Slam the door and watch } what happens :-) } } And just one more thought REALLY important: Should you ever decide to work } with a cryosectioning equipment: DANGER OF SUFFOCATION!!! Make sure that } you have a good ventilation of your room! 1 litre of liquid nitrogen (LN2) } will produce 700 litres of gaseous nitrogen (GN2). It is odourless and } tasteless and an operator in a small room may suffocate and not even } recognize that he/she is in danger!!! } You may use an oxygen analyzer (range 0 - 25%) for testing ... } } Hope that helps you, } } Joachim } } Dr. Joachim Prutsch } Product Manager EM Specimen Preparation } } Leica Microsystems GmbH } Hernalser Hauptstr. 219 email: } Joachim.Prutsch-at-leica-microsystems.com } A 1170 Vienna Tel.: +43 1 4 88 99 - 235 } AUSTRIA Fax: +43 1 4 88 99 - 350 } } } }
I have found that Accurate (don't have the address handy, somewhere in NY state if I remember) has a wide variety of oddball antibody, sera, and toxin inventory. Also the website http://www.antibodygateway.com/ is sort of a clearing house for several companies. john
On 25 Jan 2001, at 16:41, Tracey M. Pepper wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I have a researcher who needs an unusual type of gold-conj. secondary } antibody. Her primary is made in goat and the only known available } anti-goat is made in rabbit. Her tissue has cross-reactivity with } rabbit. Is there any other species (donkey, dog, Brazilian } opossum...) that anyone knows of as an anti-goat gold conjugate } secondary? Do we have to make a home-made version? Thanks! } } Tracey Pepper } Supervisor } Bessey Microscopy Facility } Iowa State University } ph: 515-294-3872 } fax: 515.294.1337 }
----- Original Message ----- } From: "Bill Chissoe" {wchiss-at-ou.edu} To: "MSA listserver" {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 25, 2001 3:40 PM
Thanks to all those who responded privately with good ideas about the chiller/circulation problem, and to those who pitched in for the good of the whole List. I've made some further investigations into my own plumbing problem (....that doesn't sound right....) and found out a couple additional facts.
1. The water currently in my system is pH 5.3. 2. The green crud all over my water filter was, indeed, copper.
So, I am having some corrosion problems, presumably because of the acid water. As somebody said, iron will be attacked first where it's available, but I found only very small traces of Fe on the filter. Perhaps there's very little iron/steel available in my particular system. Checking the acidity of fresh RO water from our source showed its pH to be 5.8-6.0, which is probably good enough. I guess I'll just institute a policy of monitoring and changing the water in the system on some kind of regular basis (semiannually?). Found no trace of biologicals on the filter so at least that doesn't seem to be an issue. Since it took at least five years (maybe 6 or 7) for the lines in the ESEM to become occluded the first time, I'll assume a modest amount of care and attention will prevent further occurrences. It could well be that much of this copper is actually coming from the cooling coils in the chiller itself - unfortunately this unit (a Neslab) has a closed-up tank that you can't see inside - to add water there's just a little neck and cap like a gas tank on a car. I think I prefer the old Haskris unit we used to have. It just had a big open tank that you could easily inspect.
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
I can recommend the MohrPro print processor. We've had it for a year and a half and it's been great. It's dry-dry, compact, fast and fairly easy to clean. I believe I bought it from EMS for around 4000$.
Maria
At 09:41 AM 1/25/01 -0500, Mary Gail Engle wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Maria Ericsson Harvard Medical School EM Facility 220 Longwood Avenue Boston, MA 02115 (617) 432 1698 maria_ericsson-at-hms.harvard.edu http://www.hms.harvard.edu/core/em.html
Dear Sam, I have a similar system and I definitly get green copper oxide flakes accumulating in my reservoir. Some of the brass fittings in my microscope water system have almost completely lost their inside flanges to corrosion. I haven't seen any iron oxide residue, but I don't know all the metals in the water system. At 07:24 PM 1/25/01 -0600, you wrote: } Fellow Microscopists: } As some one on the metallurgical side of things, I may be able to } help. Chris Jeffry is on the right track. The corrosion product from Cu/Fe } couples. such Cu pipe and Fe fittings, may be magnetite, a black, magnetic } residue or, maybe, red rust but not green Cu compounds. In such a couple, Fe } corrodes and the Cu does not. I'd bet on algae as the green material. You } biologists ought to be able to tell if it's algae. } } regards, } } Sam Purdy } Technical Center } National Steel Corp. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
THE TEXAS SOCIETY FOR MICROSCOPY “Embracing All Forms of Microscopy”
April 5-7, 2001
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THURSDAY WORKSHOP “Atomic Force Microscopy” Coordinated by Steve Zeigler FRIDAY GUEST SPEAKER Cell cycle, sperm dimorphism and sperm cell biology in flowering plants Presented by Scott D. Russell SATURDAY WORKSHOP “Cryo-Microtomy” Coordinated by Al Coritz
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After following this thread dealing with corrosion occurring in the cooling system, I've decided to add my experience. In my former position at NYSCC, I had a similar problem of corrosion products clogging the cooling system on the ion mill, which was a closed loop system combined with a Neslab chiller. Although there were two cast iron fittings used (not including the chiller unit), the tubing and most of the fittings were Cu or brass. However, the corrosion products were definitely Cu-based. Why? I examined at times the elbow and tube size reduction fittings and determined that these were mostly the problem. I surmised that both cause turbulence in the water flow which promotes erosion corrosion. This would especially be more of a problem in regions of the tubing and fittings that became hot, such as near the bottom of the diffusion pump heater as well as on the trailing side of the flow direction from the diffusion pump. I learned to first check the elbow fitting on the trailing side of the pump since it almost always got clogged by corrosion products. Thus, be aware of these transitions points in the water flow path and on the proximity to the heat load as potential areas for this problem.
David * * * * * * * * * * * * * * * David T. Hoelzer, Ph.D. Metals and Ceramics Division Oak Ridge National Laboratory Bldg. 4500S, Mail Stop 6151 P. O. Box 2008 Oak Ridge, Tennessee 37830 * (865) 574-5096 {Work} * (865) 574-0641 {Fax}
on Demetalization (the selective corrosion caused by the difference in ionization tendencies among the components of alloy). Maybe that explains why the copper pieces are floating around ... would be interested in more details though,
Best regards,
Edgar
--
________________ Dr. Edgar Voelkl Senior Development Staff Member ORNL Bldg 4515, MS 6064 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
Yes, you can do this with Low-temperature SEM. I wrote a paper on this in 1989 vanGardingen, PR, Jeffree, CE and Grace, J (1989) Variation in stomatal aperture in leaves of Avena fatua L. observed by low-temperature scanning electron microscopy. Plant Cell and Environment 12, 887-897.
I have to say that the method is not applicable to all species - just those which offer an unobstructed view of the aperture. If you need any more info on this just ask. Chris
----- Original Message ----- } From: "Beth Richardson" {beth-at-dogwood.botany.uga.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 25, 2001 6:42 PM
Dear List folks
I have a colleague who would like to take some workshops or classes in Immunohistochemistry-Microscopy techniques. She's just a beginner, so something that starts with the basics would be ideal. Does anyone have recommendations on offerings of either in the greater bay area to greater Sacramento area? I can think of a few local schools that may teach such courses, but I don't know about their policies about non-students taking such classes, especially when it involves hands-on labwork.
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Reply to: RE: Immuno gold Although I didn't really understand the problem with your antibody (rabbit and goat-gold, how could it react with the sample?), there may be a solution to not using rabbit antibodies.
If you purchase an unconjugated goat antibody that binds protein A, you could then use the protein A to visualize antibody binding. Examples of antibodies that bind protein A include those from pig, dog, guinea pig, rat IgG2c and IgG1, and any IgG2a, IgG2b immunoglobulins from mouse. I think human antibodies are also good for binding protein A but have not yet found any volunteers willing to be subjected to the repeated injections required.
Paul Webster.
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Tracey M. Pepper wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a researcher who needs an unusual type of gold-conj. secondary } antibody. Her primary is made in goat and the only known available } anti-goat is made in rabbit. Her tissue has cross-reactivity with } rabbit. Is there any other species (donkey, dog, Brazilian opossum...) } that anyone knows of as an anti-goat gold conjugate secondary? Do we have } to make a home-made version? } Thanks! } } Tracey Pepper } Supervisor } Bessey Microscopy Facility } Iowa State University } ph: 515-294-3872 } fax: 515.294.1337 } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A00246D1017A; Thu, 25 Jan 2001 22:58:10 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id QAA08032 } for dist-Microscopy; Thu, 25 Jan 2001 16:44:45 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id QAA08025 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 25 Jan 2001 } 16:44:14 -0600 (CST) } Received: from mailhub.iastate.edu (mailhub.iastate.edu [129.186.1.102]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id QAA08013 } for {microscopy-at-sparc5.microscopy.com} ; Thu, 25 Jan 2001 16:44:03 -0600 (CST) } Received: from pc12223.bot.iastate.edu (pc12223.botany.iastate.edu } [129.186.12.223]) } by mailhub.iastate.edu (8.9.3/8.9.3) with ESMTP id QAA00456 } for {microscopy-at-sparc5.microscopy.com} ; Thu, 25 Jan 2001 16:42:27 -0600 } Message-Id: {4.2.0.58.20010125163543.00952880-at-pop-1.iastate.edu} } X-Sender: tpepper-at-pop-1.iastate.edu (Unverified) } X-Mailer: QUALCOMM Windows Eudora Pro Version 4.2.0.58 } Date: Thu, 25 Jan 2001 16:41:56 -0600 } To: microscopy-at-sparc5.microscopy.com } From: "Tracey M. Pepper" {tpepper-at-iastate.edu} } Subject: Immuno gold } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii"; format=flowed } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273279280 } Status: R }
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We still use a lot of film too, but gave up on darkroom printing in favor of scanning and digital image processing some time ago. Film recording still offers significant advantages over the in-microscope CCD cameras for certain applications (we use both). Compared to darkroom printing, the scanning/digital processing/dye-sub printing route is a big cost saver and reduces generation of chemical wastes. Also, digital processing with programs like PhotoShop or CorelPaint is much faster and gives better control of the process. All the time spent and print paper wasted dodging and getting tones right certainly won't be missed. Also, digital images avoid the extra step of dealing with prints in presentations and publications.
Larry
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto: Larry.Thomas-at-pnl.gov
---------- From: Maria Ericsson Sent: Friday, January 26, 2001 7:07 AM To: Mary Gail Engle; Microscopy-at-sparc5.microscopy.com Subject: Re: black & white print processors
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Re: black and white print processors:
I can recommend the MohrPro print processor. We've had it for a year and a half and it's been great. It's dry-dry, compact, fast and fairly easy to
clean. I believe I bought it from EMS for around 4000$.
Maria
At 09:41 AM 1/25/01 -0500, Mary Gail Engle wrote:
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} -----------------------------------------------------------------------. } } } Dear listers, } Our 10 year old Ilford print processor is breaking down piece by } piece. We've about reached the point of diminishing returns and so are
} considering replacing it. Not everyone has switched to digital so we } use quite a lot of film. Can any of you recommend a good black and white } print processor? } } Thank you, } } Mary Gail Engle } Manager } Electron Microscopy & Imaging Facility } University of Kentucky
____________________________
Maria Ericsson Harvard Medical School EM Facility 220 Longwood Avenue Boston, MA 02115 (617) 432 1698 maria_ericsson-at-hms.harvard.edu http://www.hms.harvard.edu/core/em.html
Hi Ya'll, if there is a kind sole out there with a tem sample prep hole saw, that is willing into cut ~20 TEM disk from microscope slide covers, I am willing to pay for the service. Please contact me off line.
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Email: m.delcerro-at-worldnet.att.net Name: Manuel del Cerro State: New York
Question: I would very much like to see light and electron microscopy (TEM/SEM) of leaves of mosses. Could you please give me the name of a book or web site that will have these? Thanks!
I´am looking for a reference giving a protocol how to embedd (human) spermatozoa for TEM. What I need also to know is if and how I have to isolate the spermatozoa out of the ejaculate. It would be advantageous for me to have a relatively dense pellet of spermatozoa and I have no idea how much percent of the ejaculate are spermatozoa and how much is `junk`.
Tanks
Stefan
**************************** Dr. Stefan Geimer University of Cologne Botanical Institute Gyrhofstr. 15 D-50931 Cologne Germany
I have a stash of about 50 cut from coverslips that I don't need. I don't know what thickness they are, but they are probably #1 or #2. Send me your address.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Bruce Brinson [mailto:brinson-at-rice.edu] Sent: Friday, January 26, 2001 4:33 PM To: MSA Listserver Subject: 3mm glass,tem
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Hi Ya'll, if there is a kind sole out there with a tem sample prep hole saw, that is willing into cut ~20 TEM disk from microscope slide covers, I am willing to pay for the service. Please contact me off line.
Careful. Galvanic corrosion is dependent on the relatvie electronegativities of the alloys involved in the corrosion cell. Not all Fe alloys will corrode in contact with copper. I know for instance, that 304SS is about the same as copper, but a little more noble and therefore it is the copper that will corrode.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Purdy, Sam [mailto:SPurdy-at-nationalsteel.com] Sent: Thursday, January 25, 2001 8:24 PM To: Microscopy-at-sparc5.microscopy.com Subject: FW: Fun with vacuum systems
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Fellow Microscopists: As some one on the metallurgical side of things, I may be able to help. Chris Jeffry is on the right track. The corrosion product from Cu/Fe couples. such Cu pipe and Fe fittings, may be magnetite, a black, magnetic residue or, maybe, red rust but not green Cu compounds. In such a couple, Fe corrodes and the Cu does not. I'd bet on algae as the green material. You biologists ought to be able to tell if it's algae.
regards,
Sam Purdy Technical Center National Steel Corp.
} ---------- } From: Chris Jeffree } Sent: Thursday, January 2001, 4:49 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Fun with vacuum systems } } --------------------------------------------------------------- --------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } --------------------------------------------------------------- --------. } } } Maybe US plumbers are better trained, but UK heating systems are } almost always iron (pressed steel steel) radiators with copper } connecting pipes. Other metals involved are brass valves and } connectors, and lead/tin solder at soldered joints. Systems like that } corrode unless inhibitors are used. The sludge that collects in my } copper/iron central heating system is deep black, like indian ink, not } green. } Is it possible that your green sludge is not copper but algae?? } Transparent plastic tubing in the system may allow alagal growth, } provided there is some source of carbon. Bacterial/fungal } decomposition of glycol could provide that. } Chris } } ----- Original Message ----- } } From: "John J. Bozzola" {bozzola-at-siu.edu} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, January 24, 2001 9:03 PM } Subject: Re: Fun with vacuum systems } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } Hey, deja-vu all over again...... } } } } I encountered a similar (green sludge and corrosion) problem on the } } DP's and recirculating water system of our TEM several years ago. I } } assumed that it was a reaction between the iron and copper tubing } } used in the system. Old time plumbers, for example, always warned } } about directly connecting iron and copper tubing because of the } } corrosion that would occur. I KNOW that on our DP's the iron } fittings } } have copper connections. Oh well.... } } } } JB } } } } -- } } ############################################################## } } John J. Bozzola, Ph.D., Director } } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } } 750 Communications Drive - MC 4402 } } Southern Illinois University } } Carbondale, IL 62901 U.S.A. } } Phone: 618-453-3730 } } Fax: 618-453-2665 } } Email: bozzola-at-siu.edu } } Web: http://www.siu.edu/departments/shops/cem.html } } ############################################################## } } } }
I'd like to thank Tim Richardson for his comments and interest in our upcoming Food Structure and Functionality Symposium in May 2001, which was recently publicized on this list. I wanted to take advantage of this opportunity to explain a little about this fascinating area of study.
Food Structure and Functionality is the epitome of applied microscopy and imaging. It is not merely imaging but the integrating of this information with other food quality attributes especially texture. One of our fundamental beliefs is that every change in the behaviour of a food is accompanied by a structural change at a heirarchy of levels - from the macroscopic to the molecular. In other words, this is a structure/function relationship.
If you think about this a little, you can see how the quality of food can be assessed and controlled using these methods. Processed foods with certain properties can be designed by following the evolution of structure using these methods. Food Structure and Functionality covers all aspects of foods, from their plant and animal origins, from breeding, growth and management, harvest and post-harvest (plants), and finally to the processing stage.
At present, Food Structure and Functionality researchers are scattered all over the world. There are many labs which do this type of work and don't use this name. I know that there are a number of labs in Canada's Federal Department of Agriculture (Agriculture and Agri-Food Canada) which do this type of work, but there is only 1 in our department, at our Research Centre in Kentville, Nova Scotia (1 hour drive from Halifax/Dartmouth) which actually calls itself a Food Structure lab (our lab). USDA has many labs which do this research. Many of the large food companies in the world routinely use these techniques as a valuable tool - some are able to talk about what they do while others cannot. In Europe, labs using these techniques are numerous and are well supported.
Those of us who come to this field of study are either classically trained microscopists from the bio/medical sciences, or food scientists/technologists who have discovered the power of imaging techniques in their daily work. To my knowledge, there is no university which specifically offers courses in Food Structure and Functionality, but it is the basis of many challenging PhD theses.
Hopefully this explains a little about us for the curious. You can find out more about our group at the AOCS website (www.aocs.org, on the the Food Structure and Functionality Division webpage). Also, Milos Kalab, a pioneer in Food Structure research has a website called Food under the Microscope which is really worthwhile to visit at:
http://www.magma.ca/~scimat
Thanks for the opportunity of telling you all of this.
Hi Yall, I tried to send this with "thanks, 3mm glass, tem" in the subject line...guess ''thanks" got cought in the spam filter. } Wow! My thanks to the MSA list server & those of you that reponed. My need } for 3mm disk has been met faster than I got a copy of my own messsage {:o{)}
First, I am neither an expert in physics or mathematics. Unfortunately, I shot from the hip and included my own personal terminology. My understanding of the world is based in what I would call 'intuitive modeling'. In terms of modern physics, the electron does have a definable mass. However, with my aversion to the mathematics of quantum physics, I struggle to find a more intuitive understanding. The fitting of statistical descriptions to empirical data holds far less interest to me than the discovery of theoretical considerations that explain empirical data.
In doing so, I consider the electron as having no gravitational mass, but rather a mass defined at this point based solely on its energy. This I base on the simple minded concept of classical physics of particles as 'billiard balls'. In that concept, two features are required for 'mass'- spatial dimension and gravitational effects. At this point in time, I am unaware of any experiments that can demonstrate that either property can be attributed to the electron.
Instead, I consider the electron as a dimensionless bundle of energy that affects matter solely through contactless energy interactions. While Einstein managed to equate energy with matter, the matter portion of his equations gained considerable weight from the c-squared portion. That should have shown up by now in the case of the electron.
In regards to the matter at hand (no pun intended), it can not be denied that the accelerated electron carries a field. As even relativistically accelerated electrons have, of yet, not been shown to have an increased gravitational or spatial mass, I assume, probably incorrectly, that the apparent increase in mass is due to an increase in their energy. At least a portion of that increase should be in their electro-magnetic effects, and thus their field strength and their effect on nearby charges. Consider quantum explanations of the Bremstralung radiation, which is dependent on the acceleration of the electron yet relies on the action at a distance of a field effect.
On the level of flux densities involved in electron microscopes, I also assume that the field densities brought on the microscopic level by accelerated and focussed electron beams can easily approximate those on a macro level by an electrostatically charged body brought within millimeters of a surface. The matter of scale in a focussed electron beam can not be underestimated.
While Einstein was clearly successful in uniting our concepts of energy and mass, he also left clear distinctions. If one assumes that there is a gravitational mass associated with the electron, then one would have to assume an infinite gravitational field gradient if the electron is truly dimensionless. Such a gradient would essentially extend beyond the Swartzchild radius and make the electron a small 'black hole' - essentially undefinable in our current understanding. I honestly have not done the math myself, but I think that the difference in energy attributable to the electron's charge and its mass is sufficient that modern experiments should have been able to detect them.
That mass effect should become apparent when the particle is brought near the speed of light, as the gravitational mass would then be greatly amplified. But no such effect has been measured, as far as I know.
Frankly, I think we have a long way to go to explain the electron. In the mean time, I reserve the right to define my own understanding of it, although I may regret making those views public.
On Wednesday, January 24, 2001 9:11 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] wrote: } { { File: ATT00005.txt; charset = windows-1252 } }
Allen,
I am not sure I am following you. You say
"The best high energy experiments have been unable to assign a 'mass' to the electron."
To my knowledge and from a list of "fundamental constants" in my "Solid State Physics" book by Ashcroft and Mermin, the rest mass of an electron is 9.1095 x 10^(-31) kg. Just multiply that wit 1/2 v*v and you have the kinetic Energy of the electron. Of course you have to take into account relativistic effects. Or just multiply it with v and you have the momentum.
I think, you mistake the mass of the electron with that of the neutrino, which indeed they have not been able to determine, although there are indications from recent high energy physics measurements. But that's irrelevant here.
-----Original Message----- } From: Allen R. Sampson [mailto:ars-at-sem.com] Sent: Monday, January 22, 2001 4:21 AM To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com'
Hi
Am I permitted to say that we at Protrain do a range of training CD for SEM, TEM and EDX? Ready in a week or two will be a CD on Monitoring & Maintaining an Electron Microscope that will tidy up a number of the MSA mailings on SEM and TEM performance as well as instrument calibration.
As an ex service engineer, and now with our training courses, I find the magnification "problems" mentioned by your mailings to be absolutely NORMAL. If you have checked as many instruments as I have you will have seen all the "errors" mentioned.
Surely the important point in the discussions is not where the magnification is, high or low, but how consistant it is? For example if you have an old instrument that was calibrated by the manufacturer just the differences in glass plate to film thickness must make a difference but does it matter once you know? Specimen position, therefore objective lens current, is always very critical as is the high voltage level. If you monitor high voltage over a period of several hours you will detect a change so during the working day I know that instruments do change in relation to their calibration.
Try this experiment? When you switch on the high voltage in the morning insert a test specimen (holey carbon film) find true focus (no fringe) and immediately take a test picture at say 50,000kX. Leave the microscope on and take further pictures every hour BUT most important count the focus clicks (their value will be found in your instruction book), that is the correction that you make from one test picture to the next. After a few hours one would expect the high voltage to be stable and very little focus change to take place.
Now insert a grating replica and take a picture at the exact same focus you had for your last test picture. Now turn the focus back to the position that you had for your first test picture and adjust the eucentric Z to bring the sample back in focus. Measure the two pictures and see how your microscope performs. Plus or minus 5% is very good, but what answers will you get?
If you have a gas filled high voltage tank the system will settle far quicker than an oil filled tank. You are waiting for heat gained by the tank to equal the heat lost; then the system will become stable!
Have fun
Steve Chapman Senior Consultant Protrain For consultancy and training by professionals World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
I would like suggestions for any frame grabber cards, for image capture, for both light microscope (S-video) and SEM (NTSC) applications. All of our microscopes are connected to PC based Windows NT systems. We have tried several cards with varying success. I am hoping that some of you have found cards that you are happy with. Any suggestions would be appreciated.
Regards,
Brian Wajdyk Senior Electron Microscopist On Semiconductor Product Analysis Laboratory Brian.Wajdyk-at-onsemi.com
On Sat, 27 Jan 2001 16:40:42 +0100 "Stefan.Geimer" {stefan.geimer-at-uni-koeln.de} wrote:
} I´am looking for a reference giving a protocol how to embedd (human) } spermatozoa for TEM. What I need also to know is if and how I have to } isolate the spermatozoa out of the ejaculate. It would be advantageous } for me to have a relatively dense pellet of spermatozoa and I have no } idea how much percent of the ejaculate are spermatozoa and how much is } `junk`. } } Tanks } } Stefan } } } **************************** } Dr. Stefan Geimer } University of Cologne } Botanical Institute } Gyrhofstr. 15 } D-50931 Cologne } Germany } } phone: } office: +49-(0)221-470-7022 } lab: +49-(0)221-470-3795 } } fax: +49-(0)221-470-5181 } **************************** } Hope this helps. Our lab routinely embeds rabbit and horse spermatozoa. You really will need to centrifuge the ejaculate into a reasonably dense pellet. I wouldn't worry about excess "junk". We fix the ejaculate in 4% glutaraldehyde, 5% sucrose in 1M cacodylate for 24hr. Wash with buffer. Osmicate in 1% OsO4 for 1.5hr. Wash in buffer, centrifuge then embed in 2% low melting point agarose(Sigma A5030), 2% sucrose in buffer. This does very well in 1cc syringes that have been cut off at both ends and the plunger cut even with the bottom so that you can fill them to about 0.5cc and they will fit in a centrifuge tube. You can warm and mix the sperm and agarose in the syringe, keep warm and centrifuge, then push out a perfect pellet. We then use a standard dehydration and epon protocol for TEM. Sorry I don't have published reference for this. I have it as a hand-me down protocol. Jennifer } }
==================================== Jennifer Palmer ARBL Colorado State Univ Ft. Collins, CO 80523-1683 , USA voice:(970)491-1770 fax:(970)491-3557
To All, I am hoping someone can help me locate, i.e. send me a copy of, a Gary Larson Far Side cartoon which I saw several years ago. I would like to include this cartoon in an upcoming presentation I will be giving at a local university.
The cartoon showed a janitor who had entered an electron microscope lab, propped his mop up against the wall and was shown peering into the microscope to glimpse a view of an apple he had placed in the 'viewing' area. I hope this description sounds familiar to someone. Can anyone help? Although it is not my intention, this note may well start an exchange of all cartoons illustrating the amusing side of work in the microscopy lab or any lab for that matter.
So, can one of you Larson fans help me out? Thanks.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
In what way were you not satisfied with the boards you have already tried?
Resolution - NTSC is only good for about 640 pixels across. That is too low for my use. I prefer 1024. Many prefer even higher numbers. The NTSC signal just doesn't have it there to be captured.
Noise - I would think most NTSC signals would be noisy and that would show up in a single frame capture. You should be able to do some frame averaging or integration to improve that. That depends on the system.
Usability - This will be a function of software. Because of the resolution issue, I don't even have such a system anymore. Well maybe I do, sort of. We still have a couple of units from Dazzle that we use very infrequently. When we do use them, it is for the purpose of recording a video.
At 08:03 AM 1/29/2001 -0700, you wrote:
} Fellow Microscopists, } } I would like suggestions for any frame grabber cards, for image capture, } for both light microscope (S-video) and SEM (NTSC) applications. All of } our microscopes are connected to PC based Windows NT systems. We have } tried several cards with varying success. I am hoping that some of you } have found cards that you are happy with. Any suggestions would be } appreciated.
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
{!doctype html public "-//w3c//dtd html 4.0 transitional//en"} {html} {font size=-1} Here are some references you might find useful for isolate and embed spermatazoa. {/font} {p} {hr WIDTH="100%"} {br} {p} Aleman V. Trejo R. Morales E. Hernandez-Jauregui P. Delhumeau-Ongay G. A simple and rapid technique to {br} isolate enriched populations of spermatocytes and spermatids from the immature rat testis. {b} {i} Journal of Reproduction & Fertility {/i} . {/b}
{i} 54(1):67-75, 1978 Sep. {/i} {p} Baccetti B. Burrini AG. An improved method for the scanning electron microscopy of spermatozoa. {b} {i} Journal of {/i} {/b} {br} {b} {i} Microscopy {/i} . {/b} {i} 99(1):101-7, 1973 Sep. {/i} {p} Basrur PK. Ackerley CA. Reyes ER. Doig PA. Surface morphology of the spermatozoa in infertile Welsh ponies. {b} {i} Scanning Electron Microscopy {/i} . {/b} {i} (3):511-6, 1979. {/i} {br} {p} {hr WIDTH="100%"} {br} {br} {p} {font size=-1} -- {/font} {br} {font size=-1} Katja M. Wolski {/font} {br} {font size=-1} Ph.D. Student {/font} {br} {font size=-1} University of South Florida College of Medicine {/font} {br} {font size=-1} Department of Anatomy {/font} {br} {font size=-1} Andrology Laboratory {/font} {br} {font size=-1} 12901 Bruce B. Downs Blvd., MDC 6 {/font} {br} {font size=-1} Tampa, FL 33612-4799 {/font} {br} {font size=-1} Office: 813.974.6102 {/font} {br} {font size=-1} Lab: 813.974.9434 {/font} {br} {font size=-1} Fax: 813.974.2058 {/font} {br} {font size=-1} E-mail: kwolski-at-hsc.usf.edu {/font} {/html}
If anyone is interested, the general response to my question about Image Pro software(s) was that the package is nice, easy to learn and use (big plus!), and reasonably priced. There is a demo version available from Media Cybernetics (http://www.mediacy.com/). The "Express" package is pretty basic, but can collect images, do some "rudimentary" processing/measuring, and store. "Plus" is more advanced in its analysis capabilities, and "Pro" is the suped-up version.
I also heard that the Russ plug-in for Adobe Photoshop is similar to Image Pro; I've heard wonderful things about it, but haven't had a chance to play with it, yet. And I don't know how close it is to the Media Cyb. stuff.
Thanks again to everyone who responded!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
With all due respect (long pause): Isn't this thread getting a bit old?
The person who asked this question has long since had her question answered.
Doesn't this remind one of two dogs and only one tree?
Thank You,
Earl
----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Mike Bode'" {mb-at-Soft-Imaging.com} ; "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, January 29, 2001 1:55 AM
I've just faxed it to you. I enlarged it first so shrinking it should make the fax look better. If it doesn't look OK, I can either scan and e-mail or snail mail it to you.
Ron L ----- Original Message ----- } From: Gerroir, Paul J {Paul.Gerroir-at-crt.xerox.com} To: Microscopy-at-MSA. Microscopy. com (E-mail) {Microscopy-at-sparc5.microscopy.com} Sent: Monday, January 29, 2001 10:57 AM
In a message dated 1/29/01 3:55:20 PM, thoward-at-UNM.EDU writes:
} I also heard that the Russ plug-in for Adobe Photoshop is similar to Image } Pro; I've heard wonderful things about it, but haven't had a chance to } play with it, yet. And I don't know how close it is to the Media Cyb. } stuff.
I wouldn't say they are "similar" and in fact many people use the tool kit or fovea pro plug-ins with Image Pro Plus (in which they work just as well as in Photoshop). The plug-ins implement a very wide range of algorithms for processing and measurement, which no single commercial program provides. IPPlus is quite strong in image measurement and some areas of image processing, but has areas such as FFT processing and morphological processing of binary images in which it is less complete. The tool kit fills in those gaps nicely. On the other hand, the plug-ins when used with Photoshop provide only a very limited amount of automation. You can create actions that provide efficient batch processing of images, but you cannot automate stage control, acquisition, and report creation as you can with an IP+ Macro. I don't see them as competitive products, and I don't think Media Cybernetics does either. They address different needs and in some respects are complementary. To go beyond what I've tried to say here probably verges into the area of commercialism that would be inappropriate for this forum, but I did think it important to put my 2 cents worth in since the specific question was raised.
Postdoctoral Position in Transmission Electron Microscopy - Rice University
Rice University, Department of Mechanical Engineering and Materials Science, located in Houston, TX, is seeking candidates for a postdoctoral position in transmission electron microscopy of multicomponent oxide thin films. Applicants should have extensive and demonstrated experience in several areas of TEM and a strong background and interest in materials problem solving. Preference will be given to candidates with experience in high-resolution imaging and electron energy-loss spectroscopy as well as conventional diffraction contrast imaging. Facilities at Rice include a JEOL 2010, field-emission SEM and high-resolution X-ray diffractometers, as well as sample preparation. The project will be carried out in close collaboration with the University of Houston using a state-of-the-art field-emission TEM (JEOL 2010F), with annular dark-field detector, EDS and EELS capabilities. The position is available immediately. Duration about 1-2 years, salary is commensurate with qualifications. Candidates with a Ph.D. in Materials Science or Physics will be given preferred consideration. Interested candidates should send a curriculum vitae, publication list and the names and email addresses of at least three references to:
Prof. Susanne Stemmer Rice University Department of Mechanical Engineering and Materials Science MS 321 6100 Main Street Houston, TX 77005-1892 stemmer-at-rice.edu
Applicants must have proof of legal authorization to work in the United States. Rice University is an Affirmative Action/Equal Opportunity Employer.
Hi, I just want to thank all the people who wrote to me about SEM of stomata. I regret that I do not have the time to reply individually. I appreciate all the advice and references. I will try both chemical and cryo SEM (just as soon as our central facility gets a cold stage). This is such a great list! thanks again, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Hello, I have someone wanting to do some corrosion casts from parts of a rat. I was wondering if anyone has any experience with Batson's No 17 Plastic Replica and Corrosion Kit. I believe it is a methyl methacrylate monomer.
We had trouble in the past in working with resins in the SEM. They tended to melt under the beam, even after sufficient coating. So I was hoping, to get any comments about this or other resins. Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
First of all thanks for all the info on cleaning the LKB Knife boats we are using here and I will try and get them to switch to dental wax.
Secondly, I need to know where I can find NIH Image if it is still available and is it availabe for the PC now??
Thanks
Eric A. Rosen UCLA Medical Center Electron Microscopy Lab
{ { {This message is made of 100% recycled electrons} } } |"| -`^'- {_*_} ` _ , ' _|_|_ (o o) (o o) - (o)o) - (o o) ooO--(_)--Ooo--8--(_)--Ooo-ooO'(_)--Ooo-ooO--(_)--Ooo **Everyone has a photographic memory. Some just don't have film! Eric {Los Angeles, California} Go Check out my new and alomst finished webpage at: http://www.geocities.com/Yosemite/Rapids/4855/ _ /| \\\|/// \'o.O' ( o o ) ==========oOO==(_._)==OOo==============oOO==(_)==OOo========
Since you are admittedly creating your own interpretation of realitiy in these postings, it is a bit hard to understand when you depart from accepted physics whether you are doing that by intent or otherwise, but a few comments do seem to be in order:
(1) First of all, there is absolutely no need to drag either quantum mechanics or relativity into the discussion. The discussion of an electron microscope can be accomplished very nicely by assuming that an electron is a classical particle -- the only exception is when we deal with diffraction of the electron by a small aperture or other feature. We also do not generally need to invoke quantum mechanics to discuss the interaction of free electrons with matter through electromagnetic fields -- this is due to the fact that the electromagnetic forces are very long range and it matters not a bit whether the electron is a point or an extended distribution. Unless you are operating a VERY high energy electron microscope, relativistic effects are also inconsequential. In short, a late nineteenth century physicist would not have had any trouble understanding how an electron microscope operates (except for the aforementioned matter of diffraction) nor would he have had any problem with understanding how the field of the incident electron interacted with conduction electrons in the specimen (ionization would be a different story due to the atomic shells involved). So it seems to me that you are struggling to create a great deal of complexity where none needs to exist.
(2) Your remarks about the ambiguity of the mass of the electron are "remarkable". Does an electron have mass? Of course! The rest mass of an electron is well-known (9.1x10^-31kg) and confirmed in many experiments. One of the most direct ways of observing it is via the positron "annihilation" gamma rays emitted in many nuclear decay schemes. The mechanism is as follows: a positron (anti-matter version of the electron with a positive charge) is emitted in the nuclear decay. The positron quickly encounters a normal electron and they annihilate in a flash of pure energy, giving off a pair of oppositely directed gamma rays (as required by the conservation of momentum), each gamma ray carrying 511,000 eV energy, which, using E=mc^2, is just the rest mass of the electron! But you don't have to get this exotic to know that electrons have mass -- there are countless devices (such as scanning electron microscopes and TV picture tubes) which routinely manipulate their trajectories -- and they obey F=ma with the aforesaid mass. Since the behavior of electrons in the classical, quantum mechanical, and relativistic domains are all beautifully predicted by their representation as having mass (and I personally know of no experiment or theory to the contrary), why on earth would one want to invent an alternate theory?
Fred Schamber
"Allen R. Sampson" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Sorry for not getting around to this before now. } } First, I am neither an expert in physics or mathematics. Unfortunately, I } shot from the hip and included my own personal terminology. My } understanding of the world is based in what I would call 'intuitive } modeling'. In terms of modern physics, the electron does have a definable } mass. However, with my aversion to the mathematics of quantum physics, I } struggle to find a more intuitive understanding. The fitting of } statistical descriptions to empirical data holds far less interest to me } than the discovery of theoretical considerations that explain empirical } data. } } In doing so, I consider the electron as having no gravitational mass, but } rather a mass defined at this point based solely on its energy. This I } base on the simple minded concept of classical physics of particles as } 'billiard balls'. In that concept, two features are required for 'mass'- } spatial dimension and gravitational effects. At this point in time, I am } unaware of any experiments that can demonstrate that either property can be } attributed to the electron. } } Instead, I consider the electron as a dimensionless bundle of energy that } affects matter solely through contactless energy interactions. While } Einstein managed to equate energy with matter, the matter portion of his } equations gained considerable weight from the c-squared portion. That } should have shown up by now in the case of the electron. } } In regards to the matter at hand (no pun intended), it can not be denied } that the accelerated electron carries a field. As even relativistically } accelerated electrons have, of yet, not been shown to have an increased } gravitational or spatial mass, I assume, probably incorrectly, that the } apparent increase in mass is due to an increase in their energy. At least } a portion of that increase should be in their electro-magnetic effects, and } thus their field strength and their effect on nearby charges. Consider } quantum explanations of the Bremstralung radiation, which is dependent on } the acceleration of the electron yet relies on the action at a distance of } a field effect. } } On the level of flux densities involved in electron microscopes, I also } assume that the field densities brought on the microscopic level by } accelerated and focussed electron beams can easily approximate those on a } macro level by an electrostatically charged body brought within millimeters } of a surface. The matter of scale in a focussed electron beam can not be } underestimated. } } While Einstein was clearly successful in uniting our concepts of energy and } mass, he also left clear distinctions. If one assumes that there is a } gravitational mass associated with the electron, then one would have to } assume an infinite gravitational field gradient if the electron is truly } dimensionless. Such a gradient would essentially extend beyond the } Swartzchild radius and make the electron a small 'black hole' - essentially } undefinable in our current understanding. I honestly have not done the } math myself, but I think that the difference in energy attributable to the } electron's charge and its mass is sufficient that modern experiments should } have been able to detect them. } } That mass effect should become apparent when the particle is brought near } the speed of light, as the gravitational mass would then be greatly } amplified. But no such effect has been measured, as far as I know. } } Frankly, I think we have a long way to go to explain the electron. In the } mean time, I reserve the right to define my own understanding of it, } although I may regret making those views public. } } On Wednesday, January 24, 2001 9:11 AM, Mike Bode } [SMTP:mb-at-Soft-Imaging.com] wrote: } } { { File: ATT00005.txt; charset = windows-1252 } } } } Allen, } } I am not sure I am following you. You say } } "The best high energy experiments have been unable to assign a 'mass' to } the } electron." } } To my knowledge and from a list of "fundamental constants" in my "Solid } State Physics" book by Ashcroft and Mermin, the rest mass of an electron is } 9.1095 x 10^(-31) kg. Just multiply that wit 1/2 v*v and you have the } kinetic Energy of the electron. Of course you have to take into account } relativistic effects. Or just multiply it with v and you have the momentum. } } I think, you mistake the mass of the electron with that of the neutrino, } which indeed they have not been able to determine, although there are } indications from recent high energy physics measurements. But that's } irrelevant here. } } -----Original Message----- } } From: Allen R. Sampson [mailto:ars-at-sem.com] } Sent: Monday, January 22, 2001 4:21 AM } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } Subject: RE: Question } } Yes, please forgive me. My spelling is not very accurate when I am } responding so early in the morning. Your correction on Wehnelt is correct. } Please forgive any such errors in this response, as it is similar in } timing. } } The electric field of interest is not between any parts of the column, but } rather between the electrons and the sample. I really don't want to try to } attempt any quantum exercises here, but the energy impressed on the } electrons results in their increased EM field effects. The acceleration of } the electrons controlled by the electron gun configuration controls the } energy afforded the electrons. What happens after the anode does not } affect the energy of the electrons in the beam. } } The electrons are the field. The acceleration given to them by the gun } determine their acceleration, and thus their field and relativistic } effects. Their energy is determined by the fields in the gun structure } alone. Once they are given that energy, their travel through the column is } determined by the momentum they have been given. The fields they generate } as they travel through the sample surface are the direct result of the } energy they were provided with in the electron gun. } } Here's a simple challenge - define the 'kinetic' energy of a fast moving } electron. As the kinetic energy is generally defined as the mass vs. the } velocity of that mass, you may have a problem. The best high energy } experiments have been unable to assign a 'mass' to the electron. } Apparently, the only mass associated with the electron is the Einsteinium } energy-mass equivalent of its charge. If there is any increase in an } electron's mass/energy by acceleration in an applied electric field, it is } to the electrons energy. Check with SLAC's experiments. } } A fine point, but one with merit. It doesn't matter whether the distance } from anode to sample is 10 cm or 1000 cm, if the electron can travel the } distance without interference and external force. Thus the electro-magn } etic interaction of the electron with the sample is simply dependant on the } force originally impressed on the electron by the gun. } } The field present at the sample surface will thus be determined by the } acceleration given the electrons by the gun and their current density. The } higher the beam current, the greater the field flux. The smaller the area } of the electron impingement on the surface, the higher the field flux. As } we work towards smaller circuit dimensions, these electron interactions } produce higher field flux densities as the circuit dimensions come closer } to the electron beam diameters. } } You seem to want to separate the mass related kinetic effects from the } energy of the particle, but that can not be done. As the electron is } accelerated, its energy is apparently increased, rather than its mass. The } result is that the classical and relativistic effects normally related to } mass are instead seen in an increase in the energy, and thus, the field } effects of the electron with the sample. } } On Friday, January 19, 2001 10:43 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] } wrote: } } { { File: ATT00000.txt; charset = windows-1252 } } } } Thanks, Allen, for pointing out some of the issues here. By the way, It's } "Wehnelt", not "Wienault". } } You are probably right about the ESD vs. EDS name. However, it was clear to } me what the original question was about. } } Someone pointed out, that the cause of ESD (!) failure in electronic } devices } is, that the fields increase to beyond the breakdown strength of the } devices, which causes a high and sudden current to flow through the parts } where the fields are highest, which in turn heats and destroys the sample. } I } think, that is right. It's the fields that reach critical strength, which } then causes the current flow and destruction of the device. That's what I } meant when I said you get "zapped". } } Now, about the electron microscope. Obviously I don't want to pronounce, } that electron microscopy is impossible. That would be stupid, wouldn't it? } However, as you said yourself, the sample is at ground potential, or very } near it, and so is the anode. In other words, there is not electric } potential (or very little) between the anode and the sample and thus no } electric field. Since the sample is grounded, as is the rest of the } microscope, there is no electric field between those, either. The energy of } an electron in an electric field is proportional to the electric field } (actually proportional to the square of the field, if my memory serves me). } No field - no electric energy. Of course that does not mean that the } electrons don't have energy. They have quite a lot of energy, on the order } of 20keV, by "falling" through the electric field between cathode and anode } and not being stopped by the anode. Since the sample is grounded (or should } be), the electric effects of the electron beam on the grounded sample will } be cancelled. What you have to contend with is the kinetic energy (20 } keV/electron), which is transferred to the sample. } } Now, I have always talked about grounded samples. That does not mean that } electric fields cannot be generated by the electron beam within the sample. } Obviously, you put electrons in the sample and they have to move out of the } sample. If there is any kind of resistance (an oxide layer, etc.), that } will } result in charging and an electric potential. } } -----Original Message----- } } From: Allen R. Sampson [mailto:ars-at-sem.com] } Sent: Friday, January 19, 2001 2:44 AM } To: 'Mike Bode' } Subject: RE: Question } } Ok, apparently a simple physics lesson needed here. } } First of all we are talking about ESD, or Electro-Static Discharge effects } here. Not EDS, or what in this field is known as Electron Dispersive } Spectroscopy. } } Secondly, the electrons arriving at the sample surface in an SEM are } getting there with an acceleration that is roughly equivalent to the the } acceleration voltage impressed at the cathode. The sample is at, or very } near, ground potential. But the electrons in the beam are reaching the } sample at close to the accelerating voltage impressed. The grid or } "Wienault" of the electron gun is at a potential roughly +500V from the } accelerating voltage and is used to induce the electrons from the cathode } through the aperture in the anode, preventing the 'space charge' effects } around the cathode and providing the first electrostatic lens in the gun. } } The potential of the anode of an electron microscope may be at ground } potential, but the electron beam is passing through the opening in the } anode. The anode is used as the second electrostatic lens, and has little } effect on the energy of the electrons passing through it. By its charge } opposite that of the electron beam, it encourages the first crossover of } the beam, but since the beam does not directly interact with it, there is } no direct energy exchange. } } Precisely what kinetic energy can be transferred to the sample if there is } no energy differential between the electron potential at the anode and the } sample? Given your understanding of the processes involved, there can be } no discernable energy changes in the sample since the electrons have no } energy, and thus, there can be no discernable energy release from the } sample. In other words, in your view, electron microscopy is impossible. } } On Thursday, January 18, 2001 2:24 PM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] } wrote: } } } } Yes, EDS is very real and chip manufacturers go to great length (end } } expense) to avoid them. However, I think the mechanism is different: } } } } EDS means, that something collects static electricity (by walking on a } } carpet, for example), then touches something that is at a very different } } potential. That's what happens if you touch a metal after walking on that } } carpet. The enormous potential difference create strong electric field } that } } finally ionize the air and you get "zapped". The strong fields can } destroy } } the electronics. } } } } In an SEM the sample is usually grounded and does not see strong fields. } } True, the electrons are accelerated to 30 KV or more, but the sample does } } not see that, as the anode usually sits at ground level with the Cathode } } being at - 30 kV. What you need to be concerned about here is the kinetic } } energy that is transferred to the sample and can heat it up considerable. } On } } the other hand, if the sample is not grounded, there will be fields } } developing. There are also other effects, such as residual organics } getting } } "cracked" and forming a residue on the sample, but that's a different } story. } } } } Michael } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } phone: (888) FIND SIS } } (303) 234-9270 } } fax: (303) 234-9271 } } email: mailto:info-at-soft-imaging.com } } web: http://www.soft-imaging.com } } =================================== } } On Wednesday, January 24, 2001 9:11 AM, Mike Bode } [SMTP:mb-at-Soft-Imaging.com] wrote: } } { { File: ATT00005.txt; charset = windows-1252 } } } } Allen R. Sampson, Owner } Advanced Research Systems } 317 North 4th. Street } St. Charles, Illinois 60174 } voice 630.513.7093 fax 630.513.7092
You can find the NIH Image home page on: http://rsb.info.nih.gov/nih-image/ The PC version is available from Scion Corporation on: http://www.scioncorp.com
Cheers,
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-msm.cam.ac.uk
On Mon, 29 Jan 2001, Eric wrote:
} Secondly, I need to know where I can find NIH Image if it is still } available and is it availabe for the PC now??
Our laboratory owns a Polaroid DMC digital camera for acquiring digital images from a Nikon Optiphot upright optical microscope. We have the need to determine the cleanliness of polished steel samples and capture clean, accurate digital images. This is a very demanding application and the presence of any dust creates unacceptable spots on the final images.
The optics in the microscope and camera mounts have been checked multiple times and are spotless. When critical images are needed, Polaroid film is used which works wonderfully. Obviously we would prefer to capture the images electronically. On visual inspection, the camera chip itself appears to have dust on the surface. We have been told that we should not attempt to service the unit ourselves and for about five hundred dollars we could have factory service. However, we are also told that with the mechanical shutter action, the problem will recur in short order. Unfortunately, we cannot tolerate the loss of productivity taken by sending the camera in for this kind of service at frequent intervals.
Does anyone have a similar problem with this camera? Can anyone suggest a possible solution to this dilemma? I speculate that electrostatic forces may be keeping the dust in place. Bursts of canned air will not remove the dust but I would be willing to try something else. I am afraid that the camera may not be adequate for these most delicate applications and we may need to find another unit to meet our needs. Any assistance would be greatly appreciated. Please contact me with any suggestions and I will respond with the results. Thank you in advance for your help.
Michael Simko Research Manager ? Metallography U. S. Steel Research and Technology Center msimko-at-uss.com
First of all thanks for all the info on cleaning the LKB Knife boats we are using here and I will try and get them to switch to dental wax. NIH Image is now Scion image and available for the PC. You can find it here. http://www.meyerinst.com/html/scion/scion_image_windows.htm
Secondly, I need to know where I can find NIH Image if it is still available and is it availabe for the PC now??
Thanks
Eric A. Rosen UCLA Medical Center Electron Microscopy Lab John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
NIH Image is still available and you can download the program at http://www.scioncorp.com/
Zhaojie Zhang Cell and Molecular Biology Oklahoma Medical Research Foundation OKC, OK 73104
-----Original Message----- } From: Eric To: 'Microscopy-at-MSA.Microscopy.Com' Sent: 1/29/01 8:19 PM
To my Fellow Microscopists,
First of all thanks for all the info on cleaning the LKB Knife boats we are using here and I will try and get them to switch to dental wax.
Secondly, I need to know where I can find NIH Image if it is still available and is it availabe for the PC now??
Thanks
Eric A. Rosen UCLA Medical Center Electron Microscopy Lab
{ { {This message is made of 100% recycled electrons} } } |"| -`^'- {_*_} ` _ , ' _|_|_ (o o) (o o) - (o)o) - (o o) ooO--(_)--Ooo--8--(_)--Ooo-ooO'(_)--Ooo-ooO--(_)--Ooo **Everyone has a photographic memory. Some just don't have film! Eric {Los Angeles, California} Go Check out my new and alomst finished webpage at: http://www.geocities.com/Yosemite/Rapids/4855/ _ /| \\\|/// \'o.O' ( o o ) ==========oOO==(_._)==OOo==============oOO==(_)==OOo========
Although we have a digital camera for our SEM, some people still prefer Polaroids over digital images. We've been using Polaroid 55 pos/neg film. We routinely wash the negatives in running water for at least 30 minutes to remove residual traces of chemicals. According to the literature supplied with the film, the negatives should cleared with 18% sodium sulfite as soon as possible after exposure, then briefly rinsed in water. I assume that this is to stop development and clear the negative. My question is: Is sodium sulfite a necessary step or can we continue to clear with a long rinse of running water? Note: we've never seen any adverse effects from washing in water alone.
Thanks in advance for your responses.
Tom Januszewski Senior Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas Dallas, TX 75390-9039 214-648-7291 Email: tom.januszewski-at-UTSouthwestern.edu
Since we do Mercox it would be unfair for Ladd to report our comparison of Mercox with other corrosion casting materials. But having said that, the work of our internal personnel and outside researchers leads us to believe the problems incurred with other corrosion casting materials in use have not been experienced with Mercox. You may find the following links of interest,
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955
Gordon Vrololjak wrote: }
} Hello, } I have someone wanting to do some corrosion casts from parts of a rat. I } was wondering if anyone has any experience with Batson's No 17 Plastic } Replica and Corrosion Kit. I believe it is a methyl methacrylate monomer. } } We had trouble in the past in working with resins in the SEM. They tended } to melt under the beam, even after sufficient coating. So I was hoping, } to get any comments about this or other resins. } Gordon. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793
I need to prepare a plan-view sample of a ZnO thin film (~200 nm) on Si (111). I would like to etch away the Si and float the thin film on a grid. Can someone please suggest me a way of doing this? I can etch the Si away by HF/HNO3 but am afraid that in doing so, it would affect my ZnO too!. Please advice.
Many thanks Prad
Pradyumna Prabhumirashi Department of Materials Science Northwestern University Phone: (847)-491-7798 Fax: (847)-491-7820 http://vpd.ms.northwestern.edu/prad.htm
Could you brainiacs please put on your thinking caps and see what you can come up with for this problem?
A guy came to the lab asking if I knew of anything he could use as an 'epoxy resist'.
Now, here's the rest of the story:
He is making CCD's for telescopes. He wants to glue a piece of material to the face of the CCD using a very low viscosity epoxy. He puts the two together dry with thin strips of tape down two sides as spacers, then by capillary action fills the space between with epoxy and lets it cure.
Sometimes excess epoxy runs out at the ends and covers some of the bonding pads on the CCD. He wanted to know if there is something he could put on the bonding pads that would shield them from the epoxy, either repel it or protect the pads and let the epoxy separate easily from the pads after it has cured.
It struck me that this is similar to the kinds of problems some material scientists might run into when preparing TEM samples via tripod polishing, ie. how to hold a sample, but then release it later.
Some more details. The bonding pads are 100 x 300 um made of 1 um thick Al. After the gluing part, 1 mil gold wires are ultrasonically bonded to the pads. He says the pads are too small to paint anything on, thinks a spray might work. Pads can't be touched or the bonding won't work right.
I thought of super glue and acetone, ala tripod polishing. He wants to avoid acetone and things like petroleum distallates. He thinks ethanol and/or water would be OK, not sure about other solvents. I thought of wax and xylene, but he wasn't sure about that one either. He took some sucrose to try as a separation layer, he will try anything.
I suggested more careful measuring of the epoxy used so it wouldn't overflow, but there are some technical problems that limit the usefulness of this technique. He might try some physical barriers to hold back the overflow, but he would still like a way to protect the pads from the epoxy in case these fail.
So, is there anything that can be put on these pads to protect them from epoxy and be removed later to open them up for bonding? If anybody knows, one of you will.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
This note is to announce the session entitled "Museum Applications of SEM" at Scanning 2001. Anyone involved in the application of scanning electron microscopy and/or x-ray microanalysis in a museum setting (natural history, art, geology, etc) is encouraged to submit an abstract.
Scanning 2001 will be held in New York City, May 5-7. Details for abstract submission can be found at:
http://www.scanning.org/
Invited speakers are:
Frankie Jackson, Museum of the Rockies - "Dinosaur Eggshell Microstructure Visualized by Scanning Electron Microscopy"
and
Mark Wypyski, Metropolitan Museum of Art - "SEM and X-ray Microanalysis Applications for Art Materials"
All the best,
Angela
--------------------------------------------- Angela V. Klaus
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
For years we have used a Kodak Dektomatic print processor (not to be confused with the stabalization technology Ektomatic processor) for both our EM negatives and prints. Polymax chemistry is fine for both. Kodak no longer makes the Dektomatic, but a suitable replacement might be the Colex Colette Pro B&W processor. We are working with Colex to determine if this processor can handle EM film material. These processors are for relatively high volume work (we process about 1500 film images per month) and are more expensive than the MorePro machine.
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718 http://www.pathology.med.unc.edu/path/microscopy/welcome.htm
I wonder if someone could tell me rough approximate costs of:
- He-Cryo 300 kV, and Cryo 400, 200, or 120kV TEM, - with FEG and equipments for automatic collection of tilt series, CCD cameras etc.
I thank you in advance
Cheers
Peter
-- ___________________________________________________________ Peter Engelhardt, PhD, docent Assistant Professor in Molecular Genetics Unit of Electron Tomography Department of Virology Haartman Institute P.O.Box 21 (Haartmaninkatu 3) FIN-00014 University of Helsinki, Finland
Email: Peter.Engelhardt-at-Helsinki.FI URL: http://www.csc.fi/jpr/emt/engelhar/ Mobil phone: 040 811 9180
I am looking for some cartoons to illustrate what is a TEM. I would like to make a poster to show the general public what is TEM.
I would appreciate very much if anyone could send me or tell me where to get these cartoons (like a sketch of a microscope), anything related to TEM would do.
Regards
Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
I am working on the analysis of boron in lignocellulosic materials, employing an electron microprobe and synthetic WDS crystal (OV-145H). Does anyone know what the lowest reported detection limit for boron in biological matrix is? Any helpful suggestions?
Thank you.
Regards,
Glenn
Glenn M. Larkin Research Scientist School of Forestry & Wood Products Michigan Technological University Houghton, MI 49931-1295 Ph: (906) 487-3316 Fax: (906) 487-2915 e-mail: gmlarkin-at-mtu.edu
The PC version of Image is available from Scion Corp BUT there is platform independent, Java version available from NIH at http://rsb.info.nih.gov/ij/ I have used both and personally, I prefer the Java version.
Cheers
Colin Veitch
Instrumentation Scientist Late Stage Innovation Group CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} } Could you brainiacs please put on your thinking caps and see what you can } come up with for this problem? } } A guy came to the lab asking if I knew of anything he could use as an } 'epoxy resist'. } } Now, here's the rest of the story: } } He is making CCD's for telescopes. He wants to glue a piece of material to } the face of the CCD using a very low viscosity epoxy. He puts the two } together dry with thin strips of tape down two sides as spacers, then by } capillary action fills the space between with epoxy and lets it cure. } } Sometimes excess epoxy runs out at the ends and covers some of the bonding } pads on the CCD. He wanted to know if there is something he could put on } the bonding pads that would shield them from the epoxy, either repel it or } protect the pads and let the epoxy separate easily from the pads after it } has cured. } } It struck me that this is similar to the kinds of problems some material } scientists might run into when preparing TEM samples via tripod polishing, } ie. how to hold a sample, but then release it later. } } Some more details. The bonding pads are 100 x 300 um made of 1 um thick Al. } After the gluing part, 1 mil gold wires are ultrasonically bonded to the } pads. He says the pads are too small to paint anything on, thinks a spray } might work. Pads can't be touched or the bonding won't work right. } } I thought of super glue and acetone, ala tripod polishing. He wants to } avoid acetone and things like petroleum distallates. He thinks ethanol } and/or water would be OK, not sure about other solvents. I thought of wax } and xylene, but he wasn't sure about that one either. He took some sucrose } to try as a separation layer, he will try anything. } } I suggested more careful measuring of the epoxy used so it wouldn't } overflow, but there are some technical problems that limit the usefulness } of this technique. He might try some physical barriers to hold back the } overflow, but he would still like a way to protect the pads from the epoxy } in case these fail. } } So, is there anything that can be put on these pads to protect them from } epoxy and be removed later to open them up for bonding? If anybody knows, } one of you will. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
I need a couple of things for a Reichert ultracut E and wondered if anyone knows a supplier. I need a couple of chuck holders (locks chuck in place for trimming the block). Also, I need a rubber 'belt' to operate the specimen arm - one broke. Thanks in advance.
I trust that all of these EM cartoons are going to find their way onto some public access site where we can all see them - and perhaps use them to liven up our lectures !
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0) 33 260 5155 Fax +27 (0) 33 260 5776 website via:http:www.nu.ac.za Email:bruton-at-nu.unp.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
} } } "Gerroir, Paul J" {Paul.Gerroir-at-crt.xerox.com} 01/30/01 09:28PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To all who responded so generously with their emails, faxes and phone calls - Thanks!
Cheers! Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Sketch of the first two-step electron microscope with magnetic lenses (1931), designed and realized by M. Knoll and E. Ruska: http://www.scandem.lu.se/
Learn how transmission electron microscopes work: http://www.unl.edu/CMRAcfem/temoptic.htm
Study common uses of the TEM: http://www.mri.psu.edu/mcl/tem.htm
Modes of transmission electron microscope operation: http://em-outreach.sdsc.edu/web-course/Sec-I.F/Sec-I.F.html
I hope that you can use some of it.
Kind regards, Jesper
_ _ (. .) ---------------------000--(_)--000---------- Jesper Vejlø Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O.Box 49, DK-4000 Roskilde DENMARK Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/afm --------------------------------------------
-----Original Message----- } From: Yan Xin [mailto:xin-at-magnet.fsu.edu] Sent: 31. januar 2001 00:29 To: microscopy-at-sparc5.microscopy.com
Hello,
I am looking for some cartoons to illustrate what is a TEM. I would like to make a poster to show the general public what is TEM.
I would appreciate very much if anyone could send me or tell me where to get these cartoons (like a sketch of a microscope), anything related to TEM would do.
Regards
Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
Tom: It has been awhile since I used the 55p/n film, but IF memory serves me correctly, the sodium sulfite was used to both stop the development process and to harden the emulsion. We used this for years (as did many microscopists) and finally switched over to positive only polaroid (when not grabbing digital images).
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Tue, 30 Jan 2001 09:26:03 -0600, tom.januszewski-at-email.swmed.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Although we have a digital camera } for our SEM, some people still } prefer Polaroids over digital } images. We've been using Polaroid } 55 pos/neg film. We routinely wash } the negatives in running water for } at least 30 minutes to remove } residual traces of chemicals. } According to the literature } supplied with the film, the } negatives should cleared with 18% } sodium sulfite as soon as possible } after exposure, then briefly rinsed } in water. I assume that this is to } stop development and clear the } negative. My question is: Is sodium } sulfite a necessary step or can we } continue to clear with a long rinse } of running water? Note: we've never } seen any adverse effects from } washing in water alone. } } Thanks in advance for your responses. } } Tom Januszewski } Senior Electron Microscopist } Molecular and Cellular Imaging Facility } UT Southwestern Medical Center at Dallas } Dallas, TX 75390-9039 } 214-648-7291 } Email: tom.januszewski-at-UTSouthwestern.edu }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Mary: Leica took over the Reichert line of products several years ago, so getting in touch with the local Leica rep (I just saw that Energy Beam Sciences is the rep here in New England) is your best bet. If that doesn't work try the links at the MSA web site or at the Microworld Resource page http://www.mwrn.com.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Tue, 30 Jan 2001 22:21:57 -0600, mary mckee wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, Listers, } } I need a couple of things for a Reichert ultracut E and wondered if anyone } knows a supplier. I need a couple of chuck holders (locks chuck in place } for trimming the block). Also, I need a rubber 'belt' to operate the } specimen arm - one broke. Thanks in advance. } } Mary Mckee } Renal Unit } MGH } Charlestown, MA } } (617)726-3696 phone } (617)726-5669 fax } } }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
31 Jan 2001 07:58:04 PST Content-Type: text/plain Content-Disposition: inline Mime-Version: 1.0 To: Microscopy-at-sparc5.microscopy.com X-Mailer: Web Mail 3.7.1.7
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L.S.
Microscopist (34) (Msc) is looking for a new challenging working environment. In total 10 years of research experience in materialscience/(fracture)mechanics. 3 years experience in SEM/EPMA/EBSD/AFM/Image-processing as well as confocal microscopy. Currently working at University.
Interested?: mail dino-at-peterbosch.com
------------------------------------- Register for your free domain name! Plus free email and a personal portal http://www.namedemo.com
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Dear Jon, I would suggest the "Mold release spray" that I use to stop the epoxy mounting material from sticking to to the molds. Mine is by Crown, number 63223 and is called "Ready Release", "Gerneral Purpose Silicone". Hope this helps. At 01:53 PM 1/30/01 -0800, you wrote:
} Could you brainiacs please put on your thinking caps and see what you can } come up with for this problem? } } A guy came to the lab asking if I knew of anything he could use as an } 'epoxy resist'. } } Now, here's the rest of the story: } } He is making CCD's for telescopes. He wants to glue a piece of material to } the face of the CCD using a very low viscosity epoxy. He puts the two } together dry with thin strips of tape down two sides as spacers, then by } capillary action fills the space between with epoxy and lets it cure. } } Sometimes excess epoxy runs out at the ends and covers some of the bonding } pads on the CCD. He wanted to know if there is something he could put on } the bonding pads that would shield them from the epoxy, either repel it or } protect the pads and let the epoxy separate easily from the pads after it } has cured. } } It struck me that this is similar to the kinds of problems some material } scientists might run into when preparing TEM samples via tripod polishing, } ie. how to hold a sample, but then release it later. } } Some more details. The bonding pads are 100 x 300 um made of 1 um thick Al. } After the gluing part, 1 mil gold wires are ultrasonically bonded to the } pads. He says the pads are too small to paint anything on, thinks a spray } might work. Pads can't be touched or the bonding won't work right. } } I thought of super glue and acetone, ala tripod polishing. He wants to } avoid acetone and things like petroleum distallates. He thinks ethanol } and/or water would be OK, not sure about other solvents. I thought of wax } and xylene, but he wasn't sure about that one either. He took some sucrose } to try as a separation layer, he will try anything. } } I suggested more careful measuring of the epoxy used so it wouldn't } overflow, but there are some technical problems that limit the usefulness } of this technique. He might try some physical barriers to hold back the } overflow, but he would still like a way to protect the pads from the epoxy } in case these fail. } } So, is there anything that can be put on these pads to protect them from } epoxy and be removed later to open them up for bonding? If anybody knows, } one of you will. } } Thanks } } Jonathan Krupp Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Dear Tom, I have used PN 55 for many years and always used the 18% sodium sulfite bath. When you slip the negative into the sodium sulfite bath and gently swish it around, the negative clears and the brown developing jelly lifts off the negative and floats in the bath. It can then be fished out and disposed of like your other film developers. I then rinse the negative for 10 to 15 minutes in running water to remove the remaining caustic. My concern would be that if you only rinse in water, the developer goes down the drain and many jurisdictions do not allow this. At 09:26 AM 1/30/01 -0600, you wrote: } Hi, } } Although we have a digital camera } for our SEM, some people still } prefer Polaroids over digital } images. We've been using Polaroid } 55 pos/neg film. We routinely wash } the negatives in running water for } at least 30 minutes to remove } residual traces of chemicals. } According to the literature } supplied with the film, the } negatives should cleared with 18% } sodium sulfite as soon as possible } after exposure, then briefly rinsed } in water. I assume that this is to } stop development and clear the } negative. My question is: Is sodium } sulfite a necessary step or can we } continue to clear with a long rinse } of running water? Note: we've never } seen any adverse effects from } washing in water alone. } } Thanks in advance for your responses. } } Tom Januszewski Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Dear all, Is there a site that has a listing of jobs for E.M. Please respond off the list server. I don't wish to clutter up the listserver with non techinical request. -- Best Regards, Gregory Rudomen Technical Specialist Electron Microscopy 631-444-7372 Greg-at-umic.sunysb.edu ************************************************* Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. *************************************************
} } Another person to try would be Dave Paschke, The Dawson Co., } (617)484-7900. He had some parts for our Reichert MeF2 when I spoke to } him a year ago...you never know. } } Diane Ciaburri } General Dynamics } Pittsfield, MA 01235 } (413)494-2847
} your freind could try to contact Mr. Rasche in Germany: } email: mikrovid-at-gmx.de } homepage: www.mikrovid.com } } best wishes, Joachim } } } Dr. Joachim Prutsch } Product Manager EM Specimen Preparation } } Leica Microsystems } Hernalser Hauptstr. 219 email: } Joachim.Prutsch-at-leica-microsystems.com } A-1170 Vienna Tel. +43 1 48899 - 235 } Austria Fax +43 1 48899 - 350 } ---------------------- Weitergeleitet von Joachim
These are my only source for Reichart parts.
Good luck Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Tom, Here is verbiage from a Polaroid Tech Sheet on Type 55 film:
Processing the reusable negative: In order to remove the reagent layer and the anti-halation dyes, the processed negative needs to be washed in an 18% sodium sulfite solution. The salts within the solution minimize the swelling in the negative's gelatin layer that would be caused by washing in water only. Swelling can cause reticulation which would remain after the negative dries.
To prevent scratches: Negative scratch resistance can be improved by treating the processed negative(after clearing in water and sodium sulfite) in a solution of Kodak Rapid Fix with Hardener (parts A&B) for two minutes. This solution should be made up and used in accordance with Kodak's recommended mix procedures, chemical caution statements, wash times and temperatures.
Seems I only communicate when I need something. Our Life Science EM lab is considering writing a proposal for a cryo stage for our Philips CM12S. We have a range of support instrumentation such as a high pressure freezer, cryo ultramicrotomy atttachment for one of our ultramicrotomes, plus other bits and bobs. We do a lot of freeze substitution, but there are times when it would be useful in the extreme to see material that has been frozen only - not subjected to immersion in solvents, chemical fixatives, etc.
To that end, we have had discussions with one or more companies who build and/or sell cryo transfer and cryo stages or specimen holders for our research microscope. Some offer what are characterized as "additional" or "ancillary" anti contamination blades or devices that are permanently installed in the objective lens gap ( if there's room) and are used to keep contaminants from settling on the specimen (which is the coldest spot in the immediate area) . We are told that these devices necessarily cost a great deal and may not even be useable if there is too much "stuff" - like detectors, etc. in the gap or if the gap is too small.
Our 'scope has a short focal length lens called a "twin lens" (as opposed to a super twin or an ultra twin or a bio twin) and we have an EDS detector, a secondary electron detector and its bias plate, a stock anticontamination horseshoe, and the specimen holder plus the objective aperture all in the vicinity.
My questions. Finally. First, is it folly to expect excellent results using a cryo holder without the extra anticontamination blades installed? I am thinking in terms of long exposure periods such as one would experience if one were trying to learn TEM Tomography - tilt series micrographs through short intervals over the full range of the holder. Second, if the blades are installed, do they limit the use of other detectors or the movement range of the specimen or any other function that I haven't thought of. I'm hoping there are those of you out there who have the blades and can comment and those of you who only have the transfer device and holder who can relay their experiences and some hint as to the quality of image achieved and the difficulty encountered either with the system or because of it.
Ours is a multi-user facility and we are trying to add to the functionality of our instruments, not to make them less useful.
Thanks very much for any comment or observation, whether on or off list.
Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
The program for the Microscopy and Microanalysis 2001 meeting that will be held in Long Beach, California from August 4-9 promises to be an exciting one and covers many aspects of light, confocal, atomic force and electron microscopy. Highlights of the scientific meeting will include a 2-day Pre-meeting Congress chaired by Dr. Dave Piston of Vanderbilt University entitled "Imaging Life: From Cells to Whole Animals", a number of pre-meeting workshops, and a variety of symposia in the Biological and Material sciences covering specific topics on the development and applications of a full range of microscopes and microscopy techniques.
Contributed presentations and posters are welcome for all scientific sessions. Deadline for submission of abstracts (2 page extended format) is February 15, 2001.
In addition to the scientific program the Local Arrangements Committee, chaired by Bob Koch and Zed Mason, have arranged several activities around the Long Beach area. These include the Sunday Golf Tournament, a Sunday evening reception on the Queen Mary, and a Wednesday Evening cruise around the Long Beach and Los Angeles breakwater on the motor yacht "Spirit".
Full information concerning the meeting including the instructions for submitting abstracts, registration forms and lodging, can be found by following the Microscopy and Microanalysis '01 link on the right side of the Microscopy Society of America web page (www.msa.microscopy.com). If there are any questions please contact me (Price-at-med.sc.edu) or the meeting managers (877-MSA-MAS-1).
I hope to see as many of you as possible in Long Beach.
Bob Price
Bob Price M&M 2001 Program Chair 803-733-3393 (T) 803-733-1533 (F)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm familiar with the cartoon that Paul wanted; we tried to get permission to use it for a t-shirt for the international EM meeting in Seattle a few years ago. But even though the artist lived there (and was even a neighbor of the then-current MSA president), the answer was a firm NO. Professional cartoonists make a living from their drawings and they won't like "public access". But MSA has a few talented amateurs, including its current president (hi, Ron!), who could contribute; the job will get done if someone (like you, for example) volunteers to do it!
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I'm plan to attempt to rotary shadow a motor protein using platinum on a carbon rod and then coat with carbon. At the moment, I am trying to evaporate the platinum from the carbon electrode and am having difficulties.
We have a denton vacuum LLC with a bench top turbo III high vacuum evaporator. We wind one inch of platinum (Pt) wire on a nail (0.045) and then we load this 1 inch of 0.008 Pt wire onto a 0.04 tip of a 1.5mm carbon rod. The wire is pushed toward the solid carbon rod coming from the other side and all of the loops are tight (touch each other). The "spring" of Pt wire is tightly wound around the carbon wire. We pull a vacuum to about 5 x 10-5 torr and bring the amperage to 10 amps (filament 2) wait and then increase the amperage. Then I've tried 20,22.24, 26 and 30 amps but usually somewhere between 26-30, the carbon appears to evaporate. At lower amps, less than 24, I don't see any Pt on the test paper. So at the higher amps, both carbon and pt evaporate and this means we are getting too much carbon rather than an initial Pt coating. Under these conditions, the Pt wire only covers a short distance of the narrow tip of the carbon electrode. Does anyone have any advice or experienced this problem?
Is anyone out there working with autoradiography at the TEM level? We are considering a project of radiolabelling plant cells and looking at the sites of incorporation as per Pickett-Heaps, 1968 Protoplasma 65: 181-205.
I would be interested in any feedback regarding techniques, pitfalls, references (current) etc. especially with regard to plants.
Thank you in advance,
Kim --- Dr. Kim Rensing Dept. of Botany, UBC 6270 University Blvd Vancouver, BC, Canada V6T 1Z4
I have played w/ my Nikon 990 a while. I am very impressed with the camera.
I have a metallurgical and a stereo microscope and I connect the Nikon using an eyepiece adaptor.
My question is: what is the best focus method and what is a suitable external monitor.
I would assume a manual focus set at some reasonable focal length, perhaps the distance the eye would perceive an object when viewed in the eye piece. They fine focus could be done with the LM?
An external monitor seems to be required. The manual indicates NTSC or PAL as video output. I assume that means only low resolution output, so no need to spend extra for a high resolution monitor? Also, the LCD monitor indicates 110,000 dots. Which I guess would be in the ball park of 300 lines and 400 pixels, so it is hard to image Nikon would put much technology to produce extra resolution for the video output since only a small fraction of users would ever connect to a monitor.
Has anyone out picked a monitor they are happy with?
What is a fair price range for a used ( {10 years old and in good condition) German or Swiss stereo microscope that can provide good resolution images up to 100X. I am looking to buy such a scope. Please contact me directly if you are selling a scope that fits this description.
For years and years, at least 20, we have been doing what you do-rinsing in 20C water. 20 year old negatives are just as clear as new ones with no signs of degradation.
Sam Purdy Tech Center National Steel Corp. Trenton, MI
} From: Tom Januszewski } Sent: Tuesday, January 2001, 10:26 AM } To: microscopy-at-msa.microscopy.com } Subject: Polaroid 55 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Although we have a digital camera } for our SEM, some people still } prefer Polaroids over digital } images. We've been using Polaroid } 55 pos/neg film. We routinely wash } the negatives in running water for } at least 30 minutes to remove } residual traces of chemicals. } According to the literature } supplied with the film, the } negatives should cleared with 18% } sodium sulfite as soon as possible } after exposure, then briefly rinsed } in water. I assume that this is to } stop development and clear the } negative. My question is: Is sodium } sulfite a necessary step or can we } continue to clear with a long rinse } of running water? Note: we've never } seen any adverse effects from } washing in water alone. } } Thanks in advance for your responses. } } Tom Januszewski } Senior Electron Microscopist } Molecular and Cellular Imaging Facility } UT Southwestern Medical Center at Dallas } Dallas, TX 75390-9039 } 214-648-7291 } Email: tom.januszewski-at-UTSouthwestern.edu }
You all did so well with the last question (epoxy resist) that I couldn't resist trying a new one on you.
My EH&S guy wants to know about the current knowledge regarding the disposal of HV tank oil that may be contaminated with PCB's. He has an HV tank from a Philips x-ray machine, circa 1970's, rated at 100KV. He asked me what is usually done to dispose of these tanks. The information he finds is ambiguous. In some cases if the oil is drained, it can be disposed of one way that may be cheaper than getting rid of the whole thing intact. On the other hand, if he can just get rid of the whole thing as one piece it would be a lot less mess.
Its the PCB's in the oil that complicates things. They were put in most HV oils of that vintage, primarily to reduce the flamability of the simple mineral oil used in the tank (I think). He would like to avoid an assay for PCB's just to tell him something we already know. Also, if there is really solid evidence and reasons for special care and caution with th oil, it would help him chart the best course for responsible disposal.
Thanks,
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
To second Paul Gerroir's request for the Gary Larson SEM cartoon, can anyone e-mail or FAX one to me? I had forgotten that I had it, and another Gary Larson one involving a light microscope attached to a car steering wheel as a 'cool option', on the wall near our SEM. Then we had an explosion that ruined my lab and SEM -- and the cartoons. (The lab has since been rebuilt & SEM replaced.)
Thanks!
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 FAX 978-475-2951 jlagoy-at-bodycote-imt.com
} My EH&S guy wants to know about the current knowledge regarding the } disposal of HV tank oil that may be contaminated with PCB's. He has an HV } tank from a Philips x-ray machine, circa 1970's, rated at 100KV. He asked } me what is usually done to dispose of these tanks. The information he finds } is ambiguous. In some cases if the oil is drained, it can be disposed of } one way that may be cheaper than getting rid of the whole thing intact. On } the other hand, if he can just get rid of the whole thing as one piece it } would be a lot less mess.
{snip}
We managed to talk our local electric company into taking our old oil off our hands - it was a miniscule amount for them. It was rather under the table, to avoid all the inevitable paperwork, so call in any favors you can think of... They are, however, prepared to deal with HV oil with PCBs in it.
Good luck! Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
If things appear quiet or problematic with the Listserver it is due to network problems. Connectivity will be sparse to intermittent at best for the next few days until we sort out what is wrong. As far as I can tell the problem is "outside" my local LAN. The worrying thing is that it may be somewhere under the snow and ice.
PCB's were a real concern even in the early 70's. Your oil tank may or may not have PCB's at all as I believe they were banned at that time. The procedure we used (in the early 70's) was to first confirm the PCB's by testing. Several outside labs tested for a modest fee.
I really don't know who would test nowadays. Maybe someone on the listserver would know.
Earl
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 31, 2001 2:33 PM
At Oklahoma State they have a group that disposes of what ever you have as long as you know what it is and were it came from. You don't need to know if it has in it just the source. High voltage oil would be good enough for them. They do the testing if needed. It is a lot cheaper for the university for one office to deal with it than a bunch.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 31, 2001 4:33 PM
TO: THOSE INVOLVED IN MICROSCOPY OF INVERTEBRATES
CONFERENCE ANNOUNCEMENT
The 9th International Congress on Invertebrate Reproduction & Development is to be held in South Africa (Rhodes University, Grahamstown) from July 15-20th. This integrative meeting welcomes papers on all aspects of invertebrate reproduction. The second circular with details is available at the conference web site: http://www.rhodes.ac.za/conferences/icird2001 where it is also possible to register online. The deadline date for registration and submission of abstracts is March 31st after which a late fee will apply. The current exchange rate is very much in favour of overseas delegates. With registration delegates get entrance to all conference sessions, daytime refreshments, lunches, conference literature and evening functions. Delegates can also book excursions to some of the local game parks where the "big five" can be seen in malaria-free reserves. The International Society for Invertebrate Reproduction was founded in 1975 - come to Africa to celebrate 25 years of the society. We want to know about your research on invertebrate reproduction.
If you require further information please do not hesitate to contact the conference organiser, Alan Hodgson (A.Hodgson-at-ru.ac.za)
Alan Hodgson -------------------------------------- Professor Alan Hodgson Dept. Zoology & Entomology Rhodes University Grahamstown 6140 South Africa Tel. (+46) 6038526 Fax (+46) 6224377 Cellphone 083 598 4892
Convener 9th International Conference on Invertebrate Reproduction & Development 15-20 July 2001. For information visit: http://www.rhodes.ac.za/conferences/icird2001/
} Date: Thu, 01 Feb 2001 10:25:14 +0100 } To: "Bob Price" {price-at-dcsmserver.med.sc.edu} , microscopy-at-sparc5.microscopy.com, confocal-at-listserve.acsu.buffalo.edu } From: Gareth Morgan {Gareth.Morgan-at-impi.ki.se} } Subject: EM Meeting in Stockholm } In-Reply-To: {041b81404221f11SERVICES-at-connect.med.sc.edu} } } Bob } } Thanks for the meeting announcement. Have you seen this one? } } http://www.biosci.ki.se/SCANDEM2001/ } } I hasten to add that I have no association with the conference. The conference scientific programme looks interesting and Sweden and Stockholm are worth a look. I know - I moved here 2 years ago. } } } }
"Words are the seeds of misunderstanding - use them carefully."
"Give us the wisdom to know and not to feel that not knowing is less than wisdom itself."
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
All due respect, but I'm not sure that anyone really has an answer to her question. I've had responses that said ESD effects are very similar to electron beam damage and I've had others who have said not. Frankly, I'm confused on the issue and would love some definitive response. Have one to offer?
On Monday, January 29, 2001 2:05 PM, Earl Weltmer [SMTP:earlw-at-pacbell.net] wrote: } } With all due respect (long pause): Isn't this thread getting a bit old? } } The person who asked this question has long since had her question answered. } } Doesn't this remind one of two dogs and only one tree? } } Thank You, } } Earl } } } } } Allen R. Sampson, Owner } } Advanced Research Systems } } 317 North 4th. Street } } St. Charles, Illinois 60174 } } voice 630.513.7093 fax 630.513.7092 } } } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Dear LISTERS-FRIENDS, Thank your very much to everyone for your kind replies on my problems with electron miocroscope JEM-4000EX.
There was problem in valve V4: the leak in the place of welding of internal bellow. This leak exist in open condition of valve only. We have changed this valve on new and now everything is OK. Again thank very much for help. Looking forward to help YOU in future.
Sincerely yours Anton Gutakovskii Laboratory of Electron Microscopy Institute of Semiconductor Physics Novosibirsk, Russia - mailto:gut-at-thermo.isp.nsc.ru
Can I refer you to an excellent article in Microscopy Today Issue 01-1. Page 28 (Rotary Shadowing Macromolecules) By Douglas R Keene.
drk-at-shcc.org
Hope it helps
Kind regards
Gareth Robinson
Gareth Robinson
Emitech Ltd - Serving the Science of Electron Microscopy South Stour Avenue Ashford Kent - TN23 7RS United Kingdom Tel +44 (0) 1233 646332 Fax +44 (0) 1233 640744 Email:EM-at-emitech.co.uk HTTP://www.emitech.co.uk
This communication contains information which is confidential and may also be privileged. It is for the exclusive use of the addressee. If you are not the addressee, please note that any distribution, dissemination, copying or use of this communication or the information in it is prohibited, and may be unlawful. If you have received this message in error please advise the sender
} -----Original Message----- } From: Heinrich Matthies [SMTP:hjmatthies-at-ucdavis.edu] } Sent: 31 January 2001 23:00 } To: Microscopy-at-sparc5.microscopy.com } Subject: rotary shadowing, platinum coating } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } } } Hello, } } I'm plan to attempt to rotary shadow a motor protein using platinum on } a } carbon rod and then coat with carbon. At the moment, I am trying to } evaporate the platinum from the carbon electrode and am having } difficulties. } } We have a denton vacuum LLC with a bench top turbo III high vacuum } evaporator. We wind one inch of platinum (Pt) wire on a nail (0.045) } and } then we load this 1 inch of 0.008 Pt wire onto a 0.04 tip of a 1.5mm } carbon } rod. The wire is pushed toward the solid carbon rod coming from the } other } side and all of the loops are tight (touch each other). The "spring" } of Pt } wire is tightly wound around the carbon wire. We pull a vacuum to } about 5 } x 10-5 torr and bring the amperage to 10 amps (filament 2) wait and } then } increase the amperage. Then I've tried 20,22.24, 26 and 30 amps but } usually somewhere between 26-30, the carbon appears to evaporate. At } lower } amps, less than 24, I don't see any Pt on the test paper. So at the } higher } amps, both carbon and pt evaporate and this means we are getting too } much } carbon rather than an initial Pt coating. Under these conditions, the } Pt } wire only covers a short distance of the narrow tip of the carbon } electrode. Does anyone have any advice or experienced this problem? } } Thank you, } } Heiner Matthies } UC Davis } MCB } 530-754-9051 } }
Earl ----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Earl Weltmer'" {earlw-at-pacbell.net} ; "'Mike Bode'" {mb-at-Soft-Imaging.com} ; "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 01, 2001 2:59 AM
Please post the following announcement:
POSITION AVAILABLE FOR EM TECHNICIAN The project involves localization of mRNA, cytoskeletal proteins and synaptic markers in neurons using in situ hybridization and immunogold methods. Expertise in immunogold and TEM desired. State of the art microscopy facility. Highly competitive salary and benefits.
Please send CV to:
Dr. Gary Bassell Department of Neuroscience Kennedy Center for Mental Retardation, #529 Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461 Tel 718-430-3648 FAX 718-430-2960 -- ------------------------------------ Gary Bassell, Ph.D Assistant Professor Department of Neuroscience Rose Kennedy Center for Mental Retardation, Room #529 Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461 Tel 718-430-3648 FAX 718-430-2960 http://www.aecom.yu.edu/asb/bassell/bassell.htm
I am seeking surplus TEM specimens for demonstration of EM to undergrad Biology students. We have acquired a Philips EM 300 which is operational but we would like to be able to show a range of biological preparations to our students. Currently we have no biological specimens whatsoever.
I would be grateful for any prepared grids which might be spared.
A brief description of a grid including a specimen's identity and perhaps the method of preparation would be useful.
Please reply off list if you might be able to assist. Thanks.
Darcy
Mr. Darcy Kehler, ext 3249 Lab Co-ordinator, Biology Trinity Western University 7600 Glover Rd Langley, BC Canada V2Y 1Y1 Switchboard: (604) 888-7511 Secretary: (604) 513-2043 fax (604) 513-2018
} } My EH&S guy wants to know about the current knowledge regarding the } disposal of HV tank oil that may be contaminated with PCB's. He has } an HV tank from a Philips x-ray machine, circa 1970's, rated at } 100KV. He asked me what is usually done to dispose of these tanks. } The information he finds is ambiguous. In some cases if the oil is } drained, it can be disposed of one way that may be cheaper than } getting rid of the whole thing intact. On the other hand, if he can } just get rid of the whole thing as one piece it would be a lot less } mess. } } Its the PCB's in the oil that complicates things. They were put in } most HV oils of that vintage, primarily to reduce the flamability of } the simple mineral oil used in the tank (I think). He would like to } avoid an assay for PCB's just to tell him something we already know. } Also, if there is really solid evidence and reasons for special care } and caution with th oil, it would help him chart the best course for } responsible disposal. }
You should first of all check with Philips, they are pretty together with info on that.
Electricity supply companies use a proprietary test kit to determine if there are PCBs in transformer oil, so contact your local.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello, I have a Durst Laborator 1200 varipoint enlarger. The Varicontrol 1200 frequently blows fuses. I am using a 1 amp slow blow fuse. The point source bulb is 100 watt. Can I get a replacement Varicontrol? Thank you, Sally Shrom
I would suggest looking at the power rating on the control unit. A 120-volt 100 watt bulb should pull 0.83 amps. That does not leave much margin or room for anything else. I would look at a 1.5A fuse if the unit rating permits it.
A on 120 V At 03:33 PM 2/1/2001 -0500, you wrote:
} Hello, } I have a Durst Laborator 1200 varipoint enlarger. The Varicontrol 1200 } frequently blows fuses. I am using a 1 amp slow blow fuse. The point } source bulb is 100 watt. Can I get a replacement Varicontrol? } Thank you, } Sally Shrom
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The person who asked the inital question, Ginny Harper, is a customer of mine. She called me the day after her question & I directed her to proper, knowledgable, people at Motorola. As far as I know, her question has been answered to her satisfaction.
The last two emails were not directed at you personally. Like many on this Listserver I don't read every subject & delete the irrelevent ones as I am very busy. I took note at this one because a customer of mine asked the initial question.
I was surprised at the tangent(s) that this thread has generated none of which relate to her question. I believe this forum was meant to share collective knowledge & experiences. It appears to have degenerated to a test of "Will & Egos".
I decline your invitation to comment on said "ESD effects". I am an SEM repairman & "ESD effects" is not my area of expertise: nor is the subject of "quantum mechanics". Even the experts in the field of quantum mechanics dis-agree with each other. (I know this first hand). I will comment only on the areas of expertise that I feel I am qualified to comment. I will, however, direct you to people who are more qualified to answer that question as it is their business.
Regards,
Earl Weltmer
The last two comments were ----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Earl Weltmer'" {earlw-at-pacbell.net} ; "'Mike Bode'" {mb-at-Soft-Imaging.com} ; "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 01, 2001 2:59 AM
In order to meet federal requirements, the oil should be tested for PCB content. A result of less than 50 mg/kg is considered non-PCB. A concentration that 50 mg/kg to 499 mg/kg is considered to be contaminated with PCBs. And a concentration of 500 mg/kg or greater is considered to be PCB. There is an entire code in the Federal Register that deals with the proper way to dispose of these types of oil. If the concentration is found to be 50 mg/kg or greater, the oil is considered to be Hazardous Waste, and falls within the RCRA standard. The following URL will take you to the EPA's PCB Home Page where your friend will find several links to answer his questions.
http://www.epa.gov/opptintr/pcb/
Hope this helps.
Regards, Beth Bray
Elizabeth P. Bray Plant Chemist, Central Laboratory South Carolina Electric and Gas Co. 2102 N. Lake Dr. Columbia, SC 29212
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 31, 2001 5:33 PM
I translate (x25.4) the funny measure 0.008 " to 0.2mm. For Pt/C coating the more commonly used thickness is 0.1mm wire. If you work out the cross section area of both wires you would find that you are using considerably more Pt than the 50 to 75mm of 0.1mm wire otherwise used. I suggest that you switch to the thinner wire because this would cover more of the 1.4mm carbon cylinder. Therefore this would take longer (in time) to evaporate. This is important because you need more than 5 seconds of evaporation to obtain an even coating - with the specimen rotating at perhaps 30 RPM. What do you mean: " } at lower amps, less than 24, I don't see any Pt on the test paper"? The C would be much more visible than the Pt and normally you would not see the Pt on filter paper. The test is in the looking. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, February 01, 2001 9:00 AM, Heinrich Matthies [SMTP:hjmatthies-at-ucdavis.edu] wrote: } } } Hello, } } I'm plan to attempt to rotary shadow a motor protein using platinum on a } carbon rod and then coat with carbon. At the moment, I am trying to } evaporate the platinum from the carbon electrode and am having difficulties. } } We have a denton vacuum LLC with a bench top turbo III high vacuum } evaporator. We wind one inch of platinum (Pt) wire on a nail (0.045) and } then we load this 1 inch of 0.008 Pt wire onto a 0.04 tip of a 1.5mm carbon } rod. The wire is pushed toward the solid carbon rod coming from the other } side and all of the loops are tight (touch each other). The "spring" of Pt } wire is tightly wound around the carbon wire. We pull a vacuum to about 5 } x 10-5 torr and bring the amperage to 10 amps (filament 2) wait and then } increase the amperage. Then I've tried 20,22.24, 26 and 30 amps but } usually somewhere between 26-30, the carbon appears to evaporate. At lower } amps, less than 24, I don't see any Pt on the test paper. So at the higher } amps, both carbon and pt evaporate and this means we are getting too much } carbon rather than an initial Pt coating. Under these conditions, the Pt } wire only covers a short distance of the narrow tip of the carbon } electrode. Does anyone have any advice or experienced this problem? } } Thank you, } } Heiner Matthies } UC Davis } MCB } 530-754-9051 } }
Hi, I am a student doing a Research paper on the subject ofLithography and Semiconductor Fabrication I need information on the subject of=20 Lithography....Its types specially Photolithography, Electron beam Lithograaphy, UV lithography and Xray lithography Plz help me...if u have any articleds plz send them to me... or tell me of WEB Resources that can help.. plz reply soon u980013-at-giki.edu.pk
I have used CD 34 QBend10 clone, to demonstrate endothelial cells for several years. It gave a clean picture with no problems.
3 months ago it started to pick up collagen, elastic and general connective tissue at such a high intensity that the technique became almost unusable.
I have changed the primary antibody and every other reagent involved in the technique. I have also tried changing the dilution factor and various blocks and pretreatments. Nothing has changed in our processing or fixation routines, and the problem is also occurring in blocks that had previously stained without the heavy background.
I am just about out of ideas, any thoughts would be appreciated.
Is there a place where you can purchase ready-made examples of things to look at with an SEM?
I'm just getting started with learning to use an SEM and I've been thinking that this would be helpful to have something to look at, that you already know what it should look like.
I am not familiar with technique you described, but let me comment your message a little bit. First of all, it's not clear to me: are you going to shadow your sample with Pt and then coat it with carbon or you want to perform Pt/C shadowing? I assume, you want to try the second one. Pt/C shadowing gives a less developed granularity because carbon protects Pt from aggregation/clusterization. The best solution for Pt/C shadowing - to use electron gun with special Pt/C insert. Many years ago, I did Pt/C thermal evaporation. For that purpose I did make special carbon rods 1.5 mm dia with hole. I was using hole to insert Pt wire inside the carbon rod. As long as you are using the same rod you will have highly reproducible results. Your set-Up seems to me is so tricky and less reproducible. About thickness: it's impossible to distinguish Pt from C in the mixed evaporation. Reasonable thickness will be when you just start to see the difference between exposed and unexposed to the shadowing area of paper. It will be something brownish, never black (it's too-o-o-o much!). If you are planning not only shadow your sample but see some details on it, vacuum in the range of 10-5 torr is not adequate. You, probably have to go to 2x10-6 torr at least. In general, the cleanness of the system is very important for shadowing. And the last: I would recommend you will practice a little bit with latex beads before start real experiment. The latex suspension is widely available from any EM supply vendors. You may chose the latex size correlated to your real molecules size. With latex, you will see perfectly the quality of your shadowing. Doing one-directional shadowing - you may determine the real angle of shadowing and the quality of the shadowed metal layer (for the real sample you may switch easily to the "rotary"). And the very last comment: rotary shadowing comes from DNA analysis. At that case it was necessary, because of elongated DNA shape and necessarily to trace the whole thing. For compact globular objects, there is no advantage for rotary shadowing. One or dual (perpendicular) shadowing may provide to you more information than rotary. Usually, the contrast of the rotary shadowed samples is less than for one/dual direction shadowing (at the same metal thickness). The geometry of the shadow may give you unique information about 3rd dimension of your object. I suggest you may try both.
Good luck!
Sergey
At 04:59 PM 1/31/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John, We recently disposed of a Philips EM-200 TEM (circa 1963) and 3 others in storage for parts. That left us with 8 high voltage tanks...4 in power cabinets and 4 in scope consoles. Our radiological and environmental management people, responsible for disposing of all hazardous waste on campus, checked the tanks for PCB's and then disposed of the oil and tanks. This is part of their job and there was no charge to us. Do check with your people on campus....I suspect that they also have the means to take care of the problem for you. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907 --------------------------------------
In order to meet federal requirements, the oil should be tested for PCB content. A result of less than 50 mg/kg is considered non-PCB. A concentration that 50 mg/kg to 499 mg/kg is considered to be contaminated with PCBs. And a concentration of 500 mg/kg or greater is considered to be PCB. There is an entire code in the Federal Register that deals with the proper way to dispose of these types of oil. If the concentration is found to be 50 mg/kg or greater, the oil is considered to be Hazardous Waste, and falls within the RCRA standard. The following URL will take you to the EPA's PCB Home Page where your friend will find several links to answer his questions.
http://www.epa.gov/opptintr/pcb/
Hope this helps.
Regards, Beth Bray
Elizabeth P. Bray Plant Chemist, Central Laboratory South Carolina Electric and Gas Co. 2102 N. Lake Dr. Columbia, SC 29212
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 31, 2001 5:33 PM
Walter McCrone has published a Particle Atlas that has lots of SEM images and their EDX spectra and there are also light images of the same materials, its been very useful reference for me. Its on a CD called PAE2 Particle Atlas. You can then collect the same materials and see what they look like on your systems or McCrone also sell collection of materials you could buy. I have collected samples of materials associated with the paper, printing/plating industry and EHS (office dusts) since that is what I look at most of the time . I have no interest in McCrone. I just use their stuff. Terry Ellis Hallmark Cards Inc. tellis2-at-hallmark.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: LM. CD 34 help
Gramp Skin Pathology wrote: } I have used CD 34 QBend10 clone, to demonstrate endothelial cells for } several years. It gave a clean picture with no problems. } } 3 months ago it started to pick up collagen, elastic and general connective } tissue at such a high intensity that the technique became almost unusable. } } I have changed the primary antibody and every other reagent involved in the } technique. I have also tried changing the dilution factor and various blocks } and pretreatments. Nothing has changed in our processing or fixation } routines, and the problem is also occurring in blocks that had previously } stained without the heavy background. } } I am just about out of ideas, any thoughts would be appreciated.
This problem will have a logical explanation for why your labeling method has stopped working. It will be based either on something you changed (so look VERY carefully at the protocol actually being used) or something that has happened to either the reagents or the samples. As the problem occurs with samples that previously worked well, then it seems that specimen preparation can be ruled out as a source. Therefore you must look carefully at the reagents and the treatments you perform on the sections.
If the fixation protocols, the tissues being used and the labeling protocols are all the same (ie there have been NO changes), then the question to ask is: how do you store and handle your antibodies?
If there is no doubt that the antibody storage is not the problem, then the next question is: what is the blocking agent being used and has that been changed recently?
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Gramp Skin Pathology wrote:
}
} } } } Mark Daymon } } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A9D026B501B2; Fri, 02 Feb 2001 01:11:44 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id UAA26664 } for dist-Microscopy; Thu, 1 Feb 2001 20:11:54 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id UAA26661 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 1 Feb 2001 } 20:11:24 -0600 (CST) } Received: from mta04.mail.mel.aone.net.au (mta04.mail.au.uu.net [203.2.192.84]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id UAA26654 } for {Microscopy-at-MSA.Microscopy.com} ; Thu, 1 Feb 2001 20:11:11 -0600 (CST) } Received: from computer-8 ([203.55.242.15]) by mta03.mail.mel.aone.net.au } with SMTP } id {20010202010416.XZGN19418.mta03.mail.mel.aone.net.au-at-computer-8} } for {Microscopy-at-MSA.Microscopy.com} ; } Fri, 2 Feb 2001 12:04:16 +1100 } Message-ID: {000201c08cb4$6b7c8b20$0ff237cb-at-computer-8} } From: "Gramp Skin Pathology" {grampath-at-camtech.net.au} } To: "mico bulletin board" {Microscopy-at-sparc5.microscopy.com} } Subject: LM. CD 34 help } Date: Fri, 2 Feb 2001 09:36:54 +1030 } MIME-Version: 1.0 } Content-Type: text/plain; } charset="iso-8859-1" } Content-Transfer-Encoding: 7bit } X-Priority: 3 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 4.72.3110.5 } X-MimeOLE: Produced By Microsoft MimeOLE V4.72.3110.3 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273279495 } Status: R }
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Thanks to all...
I think I'll just use the light bulb filament to see if everything works.
But, I'm interested in using this SEM to view fungi.
I'd like to start a small catalogue of these things.
This is probably on the intenet already?
----- Original Message ----- } From: "White, Woody N." {nwwhite-at-mcdermott.com} To: "'rad0'" {rden25-at-mindspring.com} Sent: Friday, February 02, 2001 7:39 AM
Royal Microscopical Society
12th International Conference on
MICROSCOPY OF SEMICONDUCTING MATERIALS
25-29 March 2001, University of Oxford, UK
******************************** Final Announcement ********************************
This international conference will focus on the latest developments in the study of the structural and electrical properties of semiconductors by the application of transmission and scanning electron microscopy, scanning probe microscopy and X-ray techniques.
The state-of-the-art in all important subject areas will be addressed, including the characterisation of bulk and thin film as-grown materials, the study of lattice defect and impurity behaviour and the investigation of advanced semiconductor processing procedures.
Special conference sessions will concentrate on recent developments in high-resolution imaging and analytical electron microscopy, advances in SEM and SPM applications, the characteristics of epitaxial layers (including III-V nitrides), quantum wells, wires and dots, the effects of device processing treatments (including, especially, advanced silicon technology) and metal-semiconductor contacts and silicides. Prominent invited speakers will introduce each topic area.
The full conference programme, together with registration information, is now available on the web site:
http://www.rms.org.uk/currentevents2.htm#MSMXII
Please contact the Royal Microscopical Society or the under- signed for any additional information.
Tony Cullis MSMXII Co-Chairman
**************** Professor Anthony G Cullis Head of Semiconductor Materials & Devices Group Dept of Electronic & Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD, UK
I would like to encourage people to attend the forthcoming Interamerican Microscopy Congress. Previous meetings have been great fun and scientifically rewarding. Details follow. Alwyn Eades
VI Interamerican Congress on Electron Microscopy
October 7 - 11 2001 Hotel Emporio Veracruz, México
The Interamerican Congress on Electron Microscopy was previously held in Merida Venezuela, 1992; Cancún, México, 1993; Cancún, México, 1995; Guayaquil Ecuador, 1997 and Margarita Island Venezuela, 1999 This is the official congress of Committee of Inter-American Societies for Electron Microscopy (CIASEM) and is a forum for microscopy in The Americas. The Congress will have plenary talks, invited talks and a poster session. Proceedings will be published.
Congress Site The congress will take place at Hotel Emporio, which has a fine location in the port of Veracruz: It is in the heart of the historic center. The tourist attractions of the port of Veracruz and surroundings range from a rich nightlife to sites of unmatched natural beauty. It was the home of the Olmeca culture and is where Hernán Cortés disembarked. There are wonderful beaches. The well-preserved archeological areas, vast tropical forests, and colonial cities make Veracruz a fascinating place to visit.
Conference Topics:
Materials Science Electron Microcopy of magnetic materials, fracture mechanics, thin films and semiconductors, materials, computer simulation, sol-gel materials, polymers, ceramics, glasses, renewable energy materials, recycling materials, surfaces characterization, corrosion, composites, and general topics in material science.
Biological Sciences In situ hybridization, immunolocalization, scanning probe microscopy in biology, pathology, microscopy and cell biology, neurobiology, and general topics in biological sciences
Short Courses and Labs: Sample preparation Scanning microscopy TEM and High Resolution Digital and Image Processing Advances in remote control electron microscopy
Exhibition Electron microscopes, sample preparation and related products will be displayed during the congress. Exhibitors in fields related to new microscopies and optical microscopies are encouraged to participate.
Call for papers Abstracts must be contained in exactly 2 pages. The first page will include only text, including title, authors, affiliation, main body of the work and references. The second page will include text, tables and figures. Use a word processor, with high quality printer, TIMES NEW ROMAN type letter in 12 points, and a single space line. Abstracts will be published in Acta Microscopica
Deadline to send abstracts is July 15, 2001.
English will be the official language of the Congress. Translation facilities will not be available.
Hotel Reservation Special rates are available at the Hotel Emporio for Congress attendees. For reservations contact the hotel directly mentioning the congress. Single rooms are US $98, Doubles $108.50, extra people $17. Suites are $116 (single) and $138 (double).
Before august 31, 2001 $ 300.00 USD Students: $ 100.00 USD After august 31, 2001 $ 325.00 USD Students: $ 125.00 USD Courses Before august 31, 2001 $ 60.00 USD After august 31, 2001 $ 80.00 USD
Chairman: Miguel José Yacamán (yacaman-at-che.utexas.edu)
Informations and Registration Dr. José Reyes-Gasga Instituto de Física, UNAM Apartado Postal 20-364, 01000 México D.F., México Phone: (525) 622-5083 Fax. (525) 622-5008 Email: jreyes-at-fenix.ifisicacu.unam.mx
Could anyone provide me with the telephone number of the Laboratory of Scanning Electron Microscopy at the Harvard Museum of Comparative Zoology? Many thanks in advance.
I have a model 903/904 AO automatic knife sharpener. I am missing the redressing bridge for reconditioning the glass plates. Does anyone have one they would like to get rid of?
Kreonite is pretty much a notable industry standard for paper and emulsion autoprocessing. These are not low cost units, however. Rebuilt ones can be found at places like:
http://www.dunningphoto.com/rebuilt.html
You can also search for other sources using the key model numbers which suit your needs.
gary g.
At 01:33 PM 1/30/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try Media Cybernetics Image Pro-Plus. Then add an NTSC frame grabber card which it supports. The Matrox and National Instruments cards are very good. with this complement, you can average, integrate, Kalman and do pixel mapping as well as image archiving from a standard TV video source.
http://www.mediacy.com
gary g.
At 07:03 AM 1/29/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Email: carlos-e-chavez-at-uiowa.edu Name: carlos e chavez,dds School: University of Iowa College of Dentistry State: Iowa
I am an graduate student currently enrolled in a Master's program in Operative Dentistry at University of Iowa, College of Dentistry. My research interest involves the evaluation of interfaces between composite resins used in dentistry and a noble (Au 51.5%, Pd 38%) and base metal alloys (NiCrBe). I have mounted a metal disc of 10mm. diameterX 2mm thick (Noble or base alloy) in Epoxy resin and followed the protocol used in dental research for metal preparation (280, 400,and 600 grit Si Carbide paper under running water, etc). Then, using a brush I painted on the metal surface delimited by a tape with a 6 mm diameter hole using a resin opaquer paste of 0.5 mm thick. Then using a mold of 2.38 mm diameter a applied a composite resin and light cured to harden it.Finally, I cleaned the excess opaque in the periphery. My question is how do you prepare this three element interface for observation under the SEM or optical microscope? How would I make the cuts so I do not disturb the bond between the three elements and examine at these interfaces under the microscope accurately?. I hope this is clear. Thank you very much. Carlos E. Chavez, DDS University of Iowa College of Dentistry
The "main" way Pt/C I thought was being evaporated today is the "bead on carbon" method, that is, where the wire is coiled up, around a pin of diameter slightly larger than the diameter of the neck on the carbon rod, and then, with some help from some good Style #5 tweezers, it is pulled onto the sharpened neck of the carbon rod. The bell jar is lowered over the carbon rods and holders, and wearing the appropriate eye protection, the power is slowly turned up, slowly heating the carbon rods and wound Pt wire. At some point the miracle happens: The Pt melts, and surface tension effects cause it to form (in an instant) a nice little "bead" (it looks like the textbook "sessile drop") firmly attached to the carbon rod once the heating is stopped. After cooling down, the rod with the drop is rotated around so that it is facing where the samples will be, the bell jar is then loaded, pumped down and evaporation of Pt/C can be done simultaneously this way.
Indeed I don't think it is possible to evaporate Pt wire **without** the formation of the sessile drop, therefore, how the wire is originally spread out (on the rod neck) does not matter!
A note not related to the original question: The sharpened "neck" should be sharpened to a diameter not more than 3/16" (4.75 mm). The method will not work as well if the neck diameter is larger than this. If you are sharpening your own rods, make sure you are using rods of higher rather than lower density, otherwise the rod will not have the mechanical properties needed to sharpen down to this diameter.
I had a phone call on Friday from a customer asking about 0.1 mm (e.g. 4 mil ) wire since a previous posting seemed to attribute to it something special (relative to 0.2 mm/8 mil) wire. I told the caller that we did not believe that to be so, that for the practice of the sessile drop method, either diameter wire would "bead up" just as quickly and easily, and the only difference between the two diameters would be one of cost, since the cost to draw the same mass in 4 ml is obviously going to be higher than that to draw 8 mil. Actually some years ago we came into some 10 mil Pt wire, and we found it could be used just as easily to make the sessile drop.
Furthermore since either diameter wire could be used to create a drop of equal size, the actual evaporation time would be independent of the diameter of the starting wire.
In the end, we came upon the novel conclusion that I would make a posting and subject my advice to the most stringent (microscopy) peer review panel in the world, namely the listserver. Am I not correct, in that the end result will be the same irrespective of whether 4 mil or 8 mil wire is used?
Chuck
Disclaimer: SPI Supplies is a long time supplier of the high purity Pt wire that is used for evaporation in electron microscopy applications.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Handbook of Microlithography, Micromachining, and Microfabrication : Microlithography by P. Rai-Choudhury (Editor) Hardcover Vol 001 (June 1997) SPIE Press; ISBN: 0819423785
This is a very thorough book which covers optical lithography, electron beam lithography, and x-ray lithography, as well as other related topics. The chapter on e-beam lithography is on the web at "http://www.cnf.cornell.edu/spiebook/toc.htm".
A list of other books that may be useful can be found at "www.jcnabity.com/booklist.htm".
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
In a message dated 2/1/2001 11:25:38 PM Mountain Standard Time, u980013-at-giki.edu.pk writes:
} Hi, } I am a student doing a Research paper on the subject ofLithography and } Semiconductor Fabrication } I need information on the subject of=20 } Lithography....Its types specially Photolithography, } Electron beam Lithograaphy, UV lithography and Xray lithography } Plz help me...if u have any articleds plz send them to me... } or tell me of WEB Resources that can help.. } plz reply soon } u980013-at-giki.edu.pk }
Email: Rassad-at-students.miami.edu Name: Rizwan Assad School: University of Miami
Question: I would like to know if I would be able to see a protein that weighs about 100 kilo daltons under an SEM. If so, where can I find resourses for the preparation of the specimen?
I ran across an original manual for an Oxford Vibratome--Model G-Catalog no. 501 and 502. Since we no longer have the vibratome, we don't need the manual. Does anyone want it?
Bob Robert R. Wise, Ph.D. Associate Professor of Plant Physiology Department of Biology and Microbiology University of Wisconsin Oshkosh Oshkosh, WI 54901 tele: (920) 424-3404 fax: (920) 424-1101 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
You can also look at www.integraltech.com for very reliable image capture cards. They have 1000's installed (we have sold that many alone). They have AGP and PCI versions. Also can handle signals from RGB to NTSC Composite.
As for software -- there is a product called A4i that is pretty good (not ours) and www.paxit.com (ours) that handle all of the archiving, measuring, Excel interface, report generation and image analysis functions you are most likely desiring. We have thousands of systems in stalled and many in semicon mnfg and QA. Both of these products have network versions that allow less expensive licenses for your work outside the fab.
Try Media Cybernetics Image Pro-Plus. Then add an NTSC frame grabber card which it supports. The Matrox and National Instruments cards are very good. with this complement, you can average, integrate, Kalman and do pixel mapping as well as image archiving from a standard TV video source.
http://www.mediacy.com
gary g.
At 07:03 AM 1/29/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
there are definitely many cards out there to acquire standard TV signals from any source that adheres to that standard. These standards were developed decades ago for TV cameras, and my personal opinion is, that they are good for moving images (as on TV), but they lack resolution and definition for still images (LM and SEM). However, since they cards are usually not very expensive, and some of them do offer integration capabilities (normally frame integration), it might be worth a shot. Also the microscopes themselves might be offering integration capabilities. You are limited to a resolution of 640x480 (NTSC) or 758x576 (PAL).
For better resolution you might want to look at other options as well. For SEMs there are interfaces available (such as our ADDA II, but there are other manufacturers also), normally available as "passive" or "active" systems. For LM, of course, there are digital cameras with better resolution and bit-depth (Video only carries about 8 bits of information, if you're lucky).
Look for systems that are open for upward expansion to keep your options open. Contact me offline if you are interested in getting more information about the way we do that.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
At 07:03 AM 1/29/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to request of preparation samples from poly(styrene)/poly(butyl acrylate) dispersion with 100 nm particles size for measurements by JEOL type of transmission electron microscope. We have got some problems with dilution of latexes and with time of staining of carbon, too. We would like to confirm a core/shell particles morphology of samples prepared by two step seeded emulsion polymerization. We have got some problems with magnification and contrast expansion of core/shell particles,too. Thank you very much for your help.
Petra Volfova, PhD. student of Department of plastics and rubber, Slovak Technical University,Bratislava,Slovak Republic.
MIDWEST MICROSCOPY AND MICROANALYSIS SOCIETY, INC. (MMMS) AFFILIATE OF THE MICROSCOPY SOCIETY OF AMERICA
MEETING ANNOUNCEMENT
JOINT MEETING WITH THE SOCIETY FOR APPLIED SPECTROSCOPY Tuesday, Feb. 13, 2001
At Unilever HPC USA, located at 3100 Golf Road, Rolling Meadows, IL. See directions for more details.
Social Hour: 5:30 pm Dinner: 6:00 pm Speaker: 7:00 pm _________________________________________________________________ The Infrared Microprobe in Production and Research By Koichi Nishikida
Abstract Unlike the visible microscope and electron microscope, which respectively use visible light and electrons to magnify the sample, the infrared microscope does not utilize infrared wavelengths to magnify the sample image. Instead, a visible microscope is modified, so that a magnified sample image is observed and, at the same time, an infrared beam is passed through the microscope to obtain an infrared spectrum of the sample. Therefore, the "IR microscope" should be better named the "IR microprobe".
Applications of the IR-microprobe cover applications from forensic } analysis to defect analysis, reverse engineering in the production industry, and medical diagnostic research.
In this talk, I will show how the IR-microprobe has contributed to quality improvements in the production industry, as well as recent research in medical applications. Briefly, I will address the evolution of this technique over the past five decades. ********************************************************************* Please make your dinner reservations for the upcoming meeting by calling (847)734-3712. Leave your name, company affiliation and the number of reservations. Please call by noon on February 9th, so that proper arrangements can be made. Dinner Cost: M3S Members: $25 M3S Students: $10 Nonmembers: $30
Directions to Unilever, Rolling Meadows, Illinois
From the Chicago Area: Kennedy Expressway to I-90 West (Rockford); I-90 West to Arlington Heights Rd.; North to Golf Rd. (2nd traffic light); Turn left (West) onto Golf Rd. for approximately 2 miles. The Unilever R&D facility is on the right, just before Hwy. 53, across from the forest preserve. Follow the entrance road all the way to the last parking lot (North lot) and enter the building through the far glass doors.
From I-290 Eisenhower Expressway: Stay on I-290 towards Rockford; Exit on Golf/Higgins Rds.; Continue North to Golf Rd.; Turn east on Golf Rd. Unilever is1/4 mile left on Golf Rd., across from the forest preserve. Follow the entrance road all the way to the last parking lot (North lot) and enter the building through the far glass doors.
From the Northern Suburbs (Route 53 South): Exit Woodfield Drive/Golf Rd. (58); Continue to first light; Turn left at first light; Continue to Golf Rd (58); Turn right (East) on Golf Rd.; Unilever is1/4 mile left on Golf Rd., across from the forest preserve. Follow the entrance road all the way to the last parking lot (North lot) and enter the building through the far glass doors.
If you have any questions about the meeting, please direct your inquiries to the phone number below. For questions about MMMS, please contact:
Dr. Damian Neuberger Senior Research Scientist Baxter Healthcare, Corp. (847) 270-5888 damian_neuberger-at-baxter.com
} Email: kmccleary54-at-hotmail.com } Name: Kevin McCleary } School: South Peace Secondary School } State: British Columbia, Canada } } Question: I am anticipating the purchase of prepared slides for an } introduction to cells in my grade ten science classes.Ý Will chromosomes be } easily viewed in smears of Drosophila (advertized as giant chromosomes)? } If not, could you suggest alternate subjects.Ý Our light microscopes are of } standard high school quality. } _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
Chuck's reply without the customary inclusion of the previous contribution is difficult to understand. I have now added my previous contribution and the original request below. The hanging drop method results largely in Pt evaporation followed by carbon. The method also requires breaking the vacuum and is therefore more trouble. As Sergey Ryazantsev in his thoughtful contribution explained (I erased that because there was no need to reply), its the simultaneous nature of Pt/C which yields the finer grain structure. Sergey also explained that the other means of achieving such fine grain is electron beam evaporation, but that requires special equipment. I explained in my previous contribution why the thinner wire is required. 30 years ago I used many meters annually of the 0.1mm Pt for freeze etching and Kleinschmitt/ DNA rotary shadowing. Nothing for it Chuck: you will need to stock 0.1mm Pt wire like all other major suppliers. Don't be overly concerned, very few people use these techniques now; in fact you could purchase your minor requirements from us! Disclaimer: its too obvious, we sell small quantities of 0.1mm Pt. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Sunday, February 04, 2001 12:43 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } The "main" way Pt/C I thought was being evaporated today is the "bead on } carbon" method, that is, where the wire is coiled up, around a pin of } diameter slightly larger than the diameter of the neck on the carbon rod, } and then, with some help from some good Style #5 tweezers, it is pulled onto } the sharpened neck of the carbon rod. The bell jar is lowered over the } carbon rods and holders, and wearing the appropriate eye protection, the } power is slowly turned up, slowly heating the carbon rods and wound Pt wire. } At some point the miracle happens: The Pt melts, and surface tension } effects cause it to form (in an instant) a nice little "bead" (it looks like } the textbook "sessile drop") firmly attached to the carbon rod once the } heating is stopped. After cooling down, the rod with the drop is rotated } around so that it is facing where the samples will be, the bell jar is then } loaded, pumped down and evaporation of Pt/C can be done simultaneously this } way. } } Indeed I don't think it is possible to evaporate Pt wire **without** the } formation of the sessile drop, therefore, how the wire is originally spread } out (on the rod neck) does not matter! } } A note not related to the original question: The sharpened "neck" should be } sharpened to a diameter not more than 3/16" (4.75 mm). The method will not } work as well if the neck diameter is larger than this. If you are } sharpening your own rods, make sure you are using rods of higher rather than } lower density, otherwise the rod will not have the mechanical properties } needed to sharpen down to this diameter. } } I had a phone call on Friday from a customer asking about 0.1 mm (e.g. 4 mil } ) wire since a previous posting seemed to attribute to it something special } (relative to 0.2 mm/8 mil) wire. I told the caller that we did not believe } that to be so, that for the practice of the sessile drop method, either } diameter wire would "bead up" just as quickly and easily, and the only } difference between the two diameters would be one of cost, since the cost to } draw the same mass in 4 ml is obviously going to be higher than that to draw } 8 mil. Actually some years ago we came into some 10 mil Pt wire, and we } found it could be used just as easily to make the sessile drop. } } Furthermore since either diameter wire could be used to create a drop of } equal size, the actual evaporation time would be independent of the diameter } of the starting wire. } } In the end, we came upon the novel conclusion that I would make a posting } and subject my advice to the most stringent (microscopy) peer review panel } in the world, namely the listserver. Am I not correct, in that the end } result will be the same irrespective of whether 4 mil or 8 mil wire is used? } } Chuck } } Disclaimer: SPI Supplies is a long time supplier of the high purity Pt wire } that is used for evaporation in electron microscopy applications. } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ==================================================
} From: Jim at ProSciTech [SMTP:jim-at-proscitech.com] Sent: Friday, February 02, 2001 9:59 AM To: 'Heinrich Matthies'; Microscopy-at-sparc5.microscopy.com
Folks,
Past sand donations from listserver subscribers helped start the MSA sand collection. This collection is freely given to educators to use with Microscopic Explorations and other educational programs. Recently, we advertised this resource to educators and the response has been overwhelming. Consequently much of the sand collection has been given away and the collection is in need of restoration. As you can guess, the most popular sands are from locations outside the United States.
So I am asking for donations from listserver subscribers, especially those outside the United States. At one point we had sand from every continent, but that is not the case any more. Please help us rebuild the MSA sand collection. Double bag your sand in sealable bags (Ziplock baggies work great) and mail your donations to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
} From your description of the specimen preparation it seems you have prepared a sample for some type of mechanical test (shear bond test?) If you need just observe an interface you can prepare samples for this purpose. Do not embed you disks in Epoxy. Treat all disk surface with with resin and composite. On low speed diamond saw cut disks in halves. Then embed them in Epoxy and polish with diamond pastes. All adhesives and composites should withstand this treatment easily. The most difficult part is polishing - glass particles of a composite filling could produce a lot of scratches, so treat you sections gently.
Good luck,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: "carlos-e-chavez-at-uiowa.edu"-at-sparc5.microscopy.com } [mailto:"carlos-e-chavez-at-uiowa.edu"-at-sparc5.microscopy.com] } Sent: Saturday, February 03, 2001 9:34 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist: Microscopy of Dental Interfaces } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } Email: carlos-e-chavez-at-uiowa.edu } Name: carlos e chavez,dds } School: University of Iowa College of Dentistry } State: Iowa } } I am an graduate student currently enrolled in a Master's program in } Operative Dentistry at University of Iowa, College of Dentistry. My } research interest involves the evaluation of interfaces } between composite } resins used in dentistry and a noble (Au 51.5%, Pd 38%) and base metal } alloys (NiCrBe). I have mounted a metal disc of 10mm. } diameterX 2mm thick } (Noble or base alloy) in Epoxy resin and followed the protocol used in } dental research for metal preparation (280, 400,and 600 grit } Si Carbide } paper under running water, etc). Then, using a brush I } painted on the metal } surface delimited by a tape with a 6 mm diameter hole using a } resin opaquer } paste of 0.5 mm thick. Then using a mold of 2.38 mm diameter } a applied a } composite resin and light cured to harden it.Finally, I } cleaned the excess } opaque in the periphery. My question is how do you prepare this three } element interface for observation under the SEM or optical microscope? } How would I make the cuts so I do not disturb the bond } between the three } elements and examine at these interfaces under the microscope } accurately?. } I hope this is clear. } Thank you very much. } Carlos E. Chavez, DDS } University of Iowa College of Dentistry } } } } } } } } } -------------------------------------------------------------- } ------------- } } }
We are about to buy a Hamamatsu Orca CCD camera to capture images, viewed with both fluorescent and transmitted light. This seems to be a popular choice of camera among microscopists. We would like to know which are the different (favorite) software packages that people are using to collect, display and analyze the data captured by an Orca camera.
Thank you.
Judy Trogadis Vision Science Research Program Toronto Western Research Institute 399 Bathurst St. Toronto, ON M5T 2S8 ph: 416-603-5088 fax: 416-603-5126 email:judy-at-uhnres.utoronto.ca
Eric Cowdrey and I have put together a web page that makes it relatively easy to find the e-mail addresses of academic colleagues, for the purpose of requesting reprints:
There are no a d v e r t i s e m e n t s or costs. If you use it, please let us know of omissions, corrections, or improvements you'd like, and forward it to your colleagues. --
Dr. Richard Gordon, Radiology, University of Manitoba, HSC Rm. GA216, 820 Sherbrook St. Winnipeg R3A 1R9 Canada Adjunct: Electrical & Computer Engineering, Exec Member: CSTB, CARRF, IEEE-EMBS phone:(204)789-3828, fax:(204)787-2080, e-mail: GordonR-at-ms.umanitoba.ca New book: The Hierarchical Genome & Differentiation Waves: Novel Unification of Development, Genetics & Evolution: http://www.wspc.com.sg/books/lifesci/2755.html Finding Reprint Authors' E-Mail Addresses: http://www.umanitoba.ca/faculties/medicine/radiology/search/searchindex.html
SECOND ANNOUNCEMENT: UBC LIVE-CELL COURSE: Please register by March 1!!
Hello all,
The faculty for the 2000 UBC 3D Live-cell Microscopy Course is now almost set .
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Ted Inoué Universal Imaging, PA o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Michael Weis Agriculture Canada
Basic info is below but you can get the entire brochure, including links to faculty home pages, at
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with Dr. Elaine Humphrey UBC BioSciences Microscopy Facility: University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 1, 2001 Deposit due April 15, 2001 Registration 5:00 - 7:00 PM Sunday, June 17, 2001 First Lecture 7:30 PM Sunday, June 17, 2001 Live-cell Course ends, noon Thursday, June 28, 2001 3D Image Processing Wksp Sat. June 30 - Monday, July 2
APPLICATIONS DUE BY MARCH 1, 2001
More information at : http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
or
REGARDING COURSE ORGANIZATION:
Contact: Prof. James B. Pawley, JBPAWLEY-at-FACSTAFF.WISC.EDU
REGARDING APPLICATIONS
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, an intensive eleven-day residential course concentrating on all aspects of 3D Microscopy of Living Cells will be held at the University of British Columbia, in June of 2001. The course includes 4 days on 2D techniques, 6 days of 3D techniques and a summary presentation. It covers basic microscopy to the highest level confocal microscopy. (A half-day Pre-course is offered for any who may need to brush up on basic optics!).
Topics include: o Quantitative 2D light microscopy o How to keep your cells alive o 3D imaging in confocal microscopy o Widefield/deconvolution techniques o Two-photon excitation microscopy o Fluorescent and backscattered light signals o Dye design, characteristics and use o Pixelation: The Nyquist Criterion o Lasers and laser tweezers o Objectives and aberrations o Scanning-systems: AODs and mirrors o Optimal pinhole size/photon efficiency o Detectors: operation and performance o Video-rate confocal imaging o Measuring ion concentrations o Display and measurement of 3D data
Morning lecture/demonstrations lead to hands-on laboratory exercises each afternoon that will utilize most of the commercial instruments currently available for 3D microscopic imaging. Students will work in groups of 3 or 4 throughout the discussion and laboratory sessions, and may complete a live-cell 3D study on their own specimens. In the first five years, over 130 students from 23 countries have attended. Last year, 11 separate, 3D microscopical workstations were available for student use under the supervision of a 17-member international faculty. We expect to have even more workstations in 2001. Including manufacturers representatives, the teacher/ student ratio will be almost 2:1.
INTERNATIONAL FACULTY
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Elaine Humphrey University of British Columbia o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Glen MacDonald University of Washington o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Michael Weis Agriculture Canada
TUITION
Course tuition is $2,150 US and includes lunches. On receipt of 50% deposit, students will receive preliminary group assignments and the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes the textbook, a binder of handouts, and tickets for the Opening Reception, the Manufacturer's Reception and the Beach Party. Accommodations and meals other than lunch are not included in the tuition fee. The Pre-course is $100 US.
APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrollment is limited to about 24 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts on request to read before the course begins and are encouraged to take the Pre-course on the afternoon of June 18.
Application forms, and other course information from this and past years, can be downloaded from the WWW site at:
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4 Phone: 1-604.822-3354 FAX: 1-604.265-5315 Email: ech-at-unixg.ubc.ca.
Application deadlines:
Application forms are due by March 1, 2000! Deadlines are early to facilitate setting up groups. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2000 to reserve your position. In general, deposit refunds are only be possible if your position can be filled from the waiting list. The remainder of the fees is due at registration.
DATES
Applications must be received by March 1, 2001 Deposit due April 15 2001 Registration 5:00 - 7:00 PM Sunday, June 17, 2001 First Lecture 7:30 PM Sunday, June 17, 2001 Live-cell Course ends, noon Thursday, June 28, 2001
TEACHING PHILOSOPHY
As befits teaching in an area at the boundary of "what is now known," lecturers have been chosen based on their expertise as scientists working in the field rather than because they all agree. They are encouraged more to be provocative than to be prosaic. Students should expect discussion in areas where differences of opinion exist.
Prior to the course, students will be organized into groups and encouraged to communicate by email/phone, about the "Living-cell" group projects that they will pursue during the course and that will be presented to the class on the last day of the course. It has been found that group interactions make best use of students' prior experience and can be very effective in teaching the practical skills covered in a hands-on course of this type.
ARRANGEMENTS FOR LIVE SAMPLES
Students must contact Dr. Elaine Humphrey to make necessary arrangements for the transport and maintenance of cell lines etc. needed for their projects. Organisms linked in any way with human disease are not permitted because of safety considerations. Transport and customs arrangements for living specimens are entirely the responsibility of the student. Students also attending the 3D Image Processing Course, may be able to analyze, process and display some of the 3D data collected from their specimens
The workshop will cover 3D image processing for measurement and display. Enrollment is limited to those attending the 3D Microscopy course. Tuition : $850 US (lunches and snacks incl.)
WHO SHOULD ATTEND?
The course is designed for biologists working with multidimensional and possibly multicolor microscopical data sets. Getting the data is only half of the battle. Image data in 3, 4 or even 5 dimensions may be difficult to store let alone analyze or display. This course is to help students understand the hardware and software aspects of this problem and give them the techniques they need to make the best use of their data.
The course is designed for biologists who need to make measurements on 3D microscopical data sets and then display the results in an effective manner. The course will be taught on SGI, Macintosh and IBM-compatible computers. A wide variety of software designed for the 3D microscopy market will be described, demonstrated and available for use.
Workshop Organizers
o Andreas Kriete Giessen University, DE o Felix Margadant University of Sydney, Au o Ping Chin Cheng State U. of New York, Buffalo o Elaine Humphrey University of British Columbia o Glen MacDonald University of Washington
PLAN OF INSTRUCTION Classes will meet from 8:30-12:00 and 1:00-6:00 with lecture-demonstrations followed immediately by hands-on laboratory sessions using a variety of workstations. Students will "learn-by-doing" with two to a machine. Lab handouts will describe some specific exercises to be performed on "canned" data sets. Facilities and supervision will be available until 11:00 PM, for students to work on their own data. -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I found that the 990 has specific applications, rather than being of a broad nature. It is not good for DIC metallurgical work since color balance is not effective. It is not bad for BF. It does do a nice job on a stereo scope. with color balanced lighting, the auto white balance will work well. The auto exposure is also very functional. For focusing, I used the AF mode with the zoom set to wide and use the scope focus for all focusing. The LCD display works fine for focusing. The critical part is to use the remote release cable. Otherwise, operation is a real pain. I typically had to take 3-5 shots to get one that was acceptable. Just a minor inconvenience. I shot at high rez, JPEG. I have not tried high rez TIFF mode.
Manual operation may be an ultimately better approach, but I have not worked much with this mode. The 990 is OK for quickie shots and proofs but for my work, I need higher pixel resolution and the ability to control gamma, and highlight/shadow regions. Also, the 990 lacks the provision for specific manual color temperature setting on a per-region basis. This is not an issue with a stereo. But it sure is with a compound scope.
gary g.
At 03:05 PM 1/31/01, you wrote:
} I have played w/ my Nikon 990 a while. I am very impressed with the camera. } } I have a metallurgical and a stereo microscope and I connect the Nikon using } an eyepiece adaptor. } } My question is: what is the best focus method and what is a suitable } external monitor. } } I would assume a manual focus set at some reasonable focal length, perhaps } the distance the eye would perceive an object when viewed in the eye piece. } They fine focus could be done with the LM? } } An external monitor seems to be required. The manual indicates NTSC or PAL } as video output. I assume that means only low resolution output, so no need } to spend extra for a high resolution monitor? Also, the LCD monitor } indicates 110,000 dots. Which I guess would be in the ball park of 300 } lines and 400 pixels, so it is hard to image Nikon would put much technology } to produce extra resolution for the video output since only a small fraction } of users would ever connect to a monitor. } } Has anyone out picked a monitor they are happy with? } } Ric } } SMARTech } 860-491-3299 } www.semguy.com } 19 Cornwall Drive } Goshen CT 06756 } } SMARTech } 860-491-3299 } www.semguy.com } 19 Cornwall Drive } Goshen CT 06756
We use small Pt/C "chips" (cylinder shaped ... about 1.5 mm diameter and 3 mm long). The chip is held between two (not sharpened) carbon rods in the evaporator. The "faces" of the rods are flat with a small hole in the center (this is where the chip is held). The covering evaporated this way has very small grain size and the process is "painless" compared to the drop method. If you are interested I can check for more details (manufacturer of the chips, composition, price etc.).
Best regards,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp ---------------------------------------------------------------------
} } Email: kmccleary54-at-hotmail.com } } Name: Kevin McCleary } } School: South Peace Secondary School } } State: British Columbia, Canada } } } } Question: I am anticipating the purchase of prepared slides for an } } introduction to cells in my grade ten science classes.Ý Will chromosomes be } } easily viewed in smears of Drosophila (advertized as giant chromosomes)? } } If not, could you suggest alternate subjects.Ý Our light microscopes are of } } standard high school quality. } } } _________________________________________________________________________ } Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
Notice you are in the radiology area, if you ever run across a copy of
'on the trail of the invisible light'
About the history of x-rays, lets us know, we need a copy for the museum's reference library....
thanks Ed Sharpe archivist for SMECC } } } Subj: Finding Reprint Authors' E-Mail Addresses } } Date: 2/5/01 7:29:02 PM US Mountain Standard Time } } From: gordonr-at-Ms.UManitoba.CA (Richard Gordon) } } To: microscopy-at-sparc5.microscopy.com } } } } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Eric Cowdrey and I have put together a web page that makes it } } relatively easy to find the e-mail addresses of academic colleagues, } } for the purpose of requesting reprints: } } } } Finding Reprint Authors' E-Mail Addresses } } http://www.umanitoba.ca/faculties/medicine/radiology/search/searchindex.html } } } } There are no a d v e r t i s e m e n t s or costs. If you use it, } } please let us know of omissions, corrections, or improvements you'd } } like,
At 08:25 AM 2/5/01 -0600, Joseph Neilly wrote: } Past sand donations from listserver subscribers helped start the MSA sand } collection. This collection is freely given to educators to use with } Microscopic Explorations and other educational programs.
What exactly happens in Activity 8 of Microscopic Explorations? Are you looking for sand with special characteristics?
Acostumbramos navegar la web en busca de sitios y páginas web interesantes, relacionados a la educación en todo el mundo, para establecer nuevos contactos y relaciones como objetivo principal.
De esta manera hemos encontrado su email y creemos interesante solicitarles, por favor, que visiten nuestra página relacionada al ámbito de la astronomía, biología, geología, oceanografia a través planetarios móviles, telescopios, sunspotters, etc, y servicios a las escuelas.
http://www.starlab.webprovider.com
De hacernos el favor de correspondernos, con gusto esperaremos sus opiniones, sugerencias y/o propuestas.
Además, si nos envia su dirección postal, le enviaremos gratuitamente completa información a todo color. Muchísimas gracias.
Ingeniero Alejandro Vega Representante de STARLAB para Latino America
starlab2000-at-ciudad.com.ar
te: 54 11 4572 5800 fx: 54 11 4545 5114
Salvador Maria del Carril 2341 - Buenos Aires (1419) - Argentina
I have a polyclonal antibody that was made against a protein that was cloned from sea anemone and then expressed in bacteria. The problem is that in TEM immunocytochemistry of sea anemone tissue, this antiserum is labelling not only my protein of interest, but also a homologous protein that occurs in an intracellular symbiont. I know that these are two separate proteins because Westerns of the sea anemone show a 32kD protein while Westerns of the symbiont show a 50kD protein. I want to tease apart which protein localizes only to the host tissue and which one localizes to the symbiont.
Does anyone know of any protocols to somehow do a subtraction so that I am left with a subpopulation of antibodies that recognize only the sea anemome protein and a separate subpopulation that recognizes only the symbiont protein? Then I could do TEM on each partner, separately.
Jodi Schwarz phone: 541-737-4358 Zoology Department email: schwarzj-at-bcc.orst.edu 3029 Cordley Hall Oregon State University Corvallis, OR 97331 _______________________________________________________________
you could make a homogenate of the symbiont and pre-absorb your antisera prior to staing the anemone.
} } } Hello - } } I have a polyclonal antibody that was made against a protein that was } cloned from sea anemone and then expressed in bacteria. The problem is } that in TEM immunocytochemistry of sea anemone tissue, this antiserum is } labelling not only my protein of interest, but also a homologous protein } that occurs in an intracellular symbiont. I know that these are two } separate proteins because Westerns of the sea anemone show a 32kD protein } while Westerns of the symbiont show a 50kD protein. I want to tease apart } which protein localizes only to the host tissue and which one localizes to } the symbiont. } } Does anyone know of any protocols to somehow do a subtraction so that I am } left with a subpopulation of antibodies that recognize only the sea anemome } protein and a separate subpopulation that recognizes only the symbiont } protein? Then I could do TEM on each partner, separately. } } Thanks so much for your help! } } Jodi Schwarz } ______________________________________________________________ } } Jodi Schwarz phone: 541-737-4358 } Zoology Department email: schwarzj-at-bcc.orst.edu } 3029 Cordley Hall } Oregon State University } Corvallis, OR 97331 } _______________________________________________________________
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have a polyclonal antibody that was made against a protein that was cloned from sea anemone and then expressed in bacteria. The problem is that in TEM immunocytochemistry of sea anemone tissue, this antiserum is labelling not only my protein of interest, but also a homologous protein that occurs in an intracellular symbiont. I know that these are two separate proteins because Westerns of the sea anemone show a 32kD protein while Westerns of the symbiont show a 50kD protein. I want to tease apart which protein localizes only to the host tissue and which one localizes to the symbiont.
Does anyone know of any protocols to somehow do a subtraction so that I am left with a subpopulation of antibodies that recognize only the sea anemome protein and a separate subpopulation that recognizes only the symbiont protein? Then I could do TEM on each partner, separately.
Thanks so much for your help!
Jodi Schwarz
Dear Jody, I would try affinity chromatography. Make two affinity columns by attaching each of the two proteins of interest to column material that is designed for this purpose. Then run the polyclonal antibody through one or the other. You can collect the ab that passes through one column, then detach the ab that sticks, then do the same with the other column (this gives ab that sticks to neither, thus should be removed, ab to the 32 kD protein which doesn't cross-react, ab to the 50 kD protein which also doesn't cross-react, and ab which reacts with both proteins). I don't have a detailed protocol, since I've never done it; I'd check Methods in Enzymology as a start. Good luck.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
A position is open for SIMS Research Assistant Professor (Ph.D. required) or Analyst (BS degree or higher required) at the North Carolina State University Analytical Instrumentation Facility (AIF).
Duties and responsibilities include: day to day operation and maintenance of a Cameca IMS-6F SIMS instrument, stylus profilometers, and sample preparation devices such as plasma metal coaters, vacuum ovens, etc; assistance with scheduling of access to and oversight of the above instrumentation and the SIMS laboratory and participation in SIMS analytical development and related research. A BS higher degree or higher is required (Ph.D. desired) in analytical chemistry or a materials related discipline (non biological) along with some hands on analytical experience in a multi-user SIMS facility and some working knowledge of SIMS. Experience in vacuum equipment and/or electronics maintenance; experience with microcomputers, both PC and Unix; and experience with analysis of semiconductor or other non biological materials a plus.
Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity and Affirmative Action Employer. ADA Accommodations: Phil Russell, prussell-at-ncsu.edu, 919-515-7501 ______________________________ Roberto Garcia NCSU/Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh NC 27695-7531 P: (919) 515-8628 F: (919) 515-6965 rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm
We've seen this problem on all of our DMC and DMCie cameras (four total). We would also be VERY interested in a permanent solution. We've had the 3 DMCie cameras since early last year. They all had to be serviced after a few months, and now they are showing the same signs again.
Gene Young Research Technologist Analytical Sciences, SMX Group The Dow Chemical Company
Michael Simko wrote:
{ {Our laboratory owns a Polaroid DMC digital camera for acquiring digital images from a Nikon Optiphot upright optical microscope. We have the need to determine the cleanliness of polished steel samples and capture clean, accurate digital images. This is a very demanding application and the presence of any dust creates unacceptable spots on the final images.
The optics in the microscope and camera mounts have been checked multiple times and are spotless. When critical images are needed, Polaroid film is used which works wonderfully. Obviously we would prefer to capture the images electronically. On visual inspection, the camera chip itself appears to have dust on the surface. We have been told that we should not attempt to service the unit ourselves and for about five hundred dollars we could have factory service. However, we are also told that with the mechanical shutter action, the problem will recur in short order. Unfortunately, we cannot tolerate the loss of productivity taken by sending the camera in for this kind of service at frequent intervals.
Does anyone have a similar problem with this camera? Can anyone suggest a possible solution to this dilemma? I speculate that electrostatic forces may be keeping the dust in place. Bursts of canned air will not remove the dust but I would be willing to try something else. I am afraid that the camera may not be adequate for these most delicate applications and we may need to find another unit to meet our needs. Any assistance would be greatly appreciated. Please contact me with any suggestions and I will respond with the results. Thank you in advance for your help.} }
Michael Simko Research Manager ? Metallography U. S. Steel Research and Technology Center msimko-at-uss.com
Responding to the message of {4.3.2.7.0.20010206075127.02837400-at-pc} from John Foust {jfoust-at-threedee.com} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } At 08:25 AM 2/5/01 -0600, Joseph Neilly wrote: } } Past sand donations from listserver subscribers helped start the MSA sand } } collection. This collection is freely given to educators to use with } } Microscopic Explorations and other educational programs. } } What exactly happens in Activity 8 of Microscopic Explorations? } Are you looking for sand with special characteristics? } } - John } When we do it with the Minnesota Microscopy Society we have a sneaker with sand stuck on the bottom, and have the students perform a "forensic investigation" to see if they can determine where the sneaker has been. They are given samples of sand from various locations and have to match them by color, shape, grain size, shell content etc. Depending on the age range it may help to motivate them by saying the sneaker came from a pirate who buried some treasure and we want to find it. We typically have half a dozen samples to compare with - Florida beach, White Sands NM, Duluth MN, California beach, Hawaii beach and Connecticut beach amongst others. You can see examples of them at our web site http://resolution.umn.edu/MMS/ProjectMicro/ (if you get the TV program "Popular Mechanics for Kids" look for our desert sand images there too).
There are other ways to use the sand, but getting them to look carefully at each one and describe it carefully is the overall aim.
Stuart
__________________ Stuart McKernan stuartm-at-tc.umn.edu Office:(612) 626-7594 IT Characterization Facility, University of Minnesota Desk:(612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Fax:(612) 625-5368
My boss asked me if I could forward this to all of the listers. If you are interested, you call call him or myself. Jo Dee Fish
Dear all,
We are offering our confocal laser scanning confocal microscope LSM-410 (Zeiss), equipped with:
1. internal 543 nm and external 488 and 514 nm lasers 2. Great set of optics ( air, water, glycerol, and oil immersion lenses) 3. New Pentium computer.
If you are interested in this instrument let me know.
-- Edward Monosov, Ph.D.
Director Cell Analysis & Histology Facility The BURNHAM INSTITUTE 10901 N. Torrey Pines Rd, La Jolla, CA 92037 Tel: (858) 646-3100 Office (r.#5144): ext. x3206 MP Confocal Microscopy (r.#5105): ext. x3466 Fluorescent Microscopy (r.#5118): ext. x3469 Electron Microscopy (r.#5121 & ##5123): ext. x3686
The Analytical Microscopy Group at the National Renewable Energy Laboratory (NREL) in Golden Colorado, is seeking applicants for a research associate position. The main responsibility is to carry out TEM, HREM, and EELS characterization of epitaxial and polycrystalline semiconductor thin films for photovoltaic applications. Current research topics include: 1. Defect generation and reduction using lateral epitaxial overgrowth, and 2. The microstucture, chemistry and electrical behavior of extended defects in semiconductors.
A Ph.D. in materials science or a related field, and a strong background in transmission electron microscopy are required. Experience with EELS and HREM image simulation is highly desirable. Good communication skills (verbal and written ) are essential.
Interested individuals should submit a resume, three selected publications, and the names of three references to:
Dr. Mowafak Al-Jassim NREL 1617 Cole Blvd. Golden, CO 80401 Fax: (303) 384-6446 e-mail: mo-at-nrel.gov
} At 08:25 AM 2/5/01 -0600, Joseph Neilly wrote: } } Past sand donations from listserver subscribers helped start the MSA sand } } collection. This collection is freely given to educators to use with } } Microscopic Explorations and other educational programs. } } What exactly happens in Activity 8 of Microscopic Explorations? } Are you looking for sand with special characteristics? } } - John -
"Microscopic Explorations" is the teacher's manual for Project MICRO, MSA's middle school outreach program. It teaches observation more than it teaches microscopy, and the message that sand delivers in that context is that sand is DIFFERENT. Students look at it, describe it, and locate its point of origin on a globe; further inquiry is encouraged. It can lead to a geography lesson, which is why Joe has run out of sand from other continents. Or geology, crystallography, malacology, whatever. So ANY "special characteristics" are a real plus for a creative teacher. Please donate, and provide whatever information you can, in brief form. I don't think he has any industrial sands; glassmakers and others, please note!
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
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Reply to: RE: Antibody subtraction? I have a polyclonal antibody that was made against a protein that was } cloned from sea anemone and then expressed in bacteria. The problem is } that in TEM immunocytochemistry of sea anemone tissue, this antiserum is } labelling not only my protein of interest, but also a homologous protein } that occurs in an intracellular symbiont. I know that these are two } separate proteins because Westerns of the sea anemone show a 32kD protein } while Westerns of the symbiont show a 50kD protein. I want to tease apart } which protein localizes only to the host tissue and which one localizes to } the symbiont. } } Does anyone know of any protocols to somehow do a subtraction so that I am } left with a subpopulation of antibodies that recognize only the sea anemome } protein and a separate subpopulation that recognizes only the symbiont } protein? Then I could do TEM on each partner, separately. }
A simple thing to try is to take Western blots of the sea anemone and cut up the band where you know the symbiont protein is present (buyt excluding the sea anemone protein). Incubate this membrane fragment with your diluted antibody. Anti-symbiont antibody binds to the blotting membrane leaving the antibodies to the sea anemone protein in suspension. The adsorbed antibody can be applied directly to your sample. It may work after one incubation, or you may need multiple exposure to the blotting membrane. You can of course use only one membrane fragment because you can strip off bound antibody after each use. To examine the symbiont protein, use the part of the Western with the sea anemone protein as the adsorbant.
You could also incubate the diluted antibody with homogenized bacteria (the ones in which the protein was expressed, and which brobably contaminated the initial antigen preparation). I bet that removes the extra band too.
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
JODI SCHWARZ wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello - } } I have a polyclonal antibody that was made against a protein that was } cloned from sea anemone and then expressed in bacteria. The problem is } that in TEM immunocytochemistry of sea anemone tissue, this antiserum is } labelling not only my protein of interest, but also a homologous protein } that occurs in an intracellular symbiont. I know that these are two } separate proteins because Westerns of the sea anemone show a 32kD protein } while Westerns of the symbiont show a 50kD protein. I want to tease apart } which protein localizes only to the host tissue and which one localizes to } the symbiont. } } Does anyone know of any protocols to somehow do a subtraction so that I am } left with a subpopulation of antibodies that recognize only the sea anemome } protein and a separate subpopulation that recognizes only the symbiont } protein? Then I could do TEM on each partner, separately. } } Thanks so much for your help! } } Jodi Schwarz } ______________________________________________________________ } } Jodi Schwarz phone: 541-737-4358 } Zoology Department email: schwarzj-at-bcc.orst.edu } 3029 Cordley Hall } Oregon State University } Corvallis, OR 97331 } _______________________________________________________________ } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A7AE4045027E; Tue, 06 Feb 2001 17:41:02 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA02467 } for dist-Microscopy; Tue, 6 Feb 2001 11:12:44 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA02462 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 6 Feb 2001 } 11:12:13 -0600 (CST) } Received: from ava.bcc.orst.edu (ava.BCC.ORST.EDU [128.193.86.4]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id LAA02454 } for {Microscopy-at-sparc5.microscopy.com} ; Tue, 6 Feb 2001 11:12:02 -0600 (CST) } Received: from jodi (Schwarz-3126--Z.CORDLEY.ORST.EDU [128.193.86.186]) } by ava.bcc.orst.edu (8.8.8+Sun/8.8.7) with SMTP id JAA25178 } for {Microscopy-at-sparc5.microscopy.com} ; Tue, 6 Feb 2001 09:09:51 -0800 (PST) } Message-Id: {3.0.3.32.20010206091020.006c6e60-at-bcc.orst.edu} } X-Sender: schwarzj-at-bcc.orst.edu } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.3 (32) } Date: Tue, 06 Feb 2001 09:10:20 -0800 } To: Microscopy-at-sparc5.microscopy.com } From: JODI SCHWARZ {schwarzj-at-bcc.orst.edu} } Subject: Antibody subtraction? } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273279589 } Status: U } Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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Dear Jody, I would try affinity chromatography. Make two affinity columns by attaching each of the two proteins of interest to column material that is designed for this purpose. Then run the polyclonal antibody through one or the other. You can collect the ab that passes through one column, then detach the ab that sticks, then do the same with the other column (this gives ab that sticks to neither, thus should be removed, ab to the 32 kD protein which doesn't cross-react, ab to the 50 kD protein which also doesn't cross-react, and ab which reacts with both proteins). I don't have a detailed protocol, since I've never done it; I'd check Methods in Enzymology as a start. Good luck.
Yours,
Bill Tivol
Reply: Hi Bill,
Although an accepted method for antibody purification, there are a couple of problems with affinity purification. One is that there has to be enough purified antigen (protein) available to put onto a column. It is increasingly clear that many antigens are just not that abundant and so other methods have to be found to relace this form of purification. Small strips of nitrocellulose into which has been embedded a small amount of the protein (Western blotting), purified by gel electrophoresis, are very useful for small volumes. These strips are useful both for affinity purification of small amounts of antigen, and for subtractive adsorption. This is where antigens are added to antibodies to remove specific binding species. This leaves the specific antibodies of interest in suspension.
The other main problem with affinity chromatography is more theoretical than real. Antibody binding to antigens is variable. Some antibodies will have high affinity and bind so tightly that it is almost impossible to remove them from the antigen. If all antibodies to be used for immunocytochemistry go through an affinity purification step it is logical to assume that many of the very best antibodies will have been removed before we even start to label with them.
As I said, this is more a theoretical problem I put forward than one I have experienced. I have not researched studies that may have observed this effect in practice, but I am sure it could possibly occur. Has anyone seen or heard about this effect occurring in practice?
Regards,
Paul Webster.
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
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I am seeking a device which will provide live NTSC frame integration or frame averaging for my full TV rate SEM. I would prefer a "stand alone box", but a PC card with a live-pass through would be OK.
I already own an Optical Electronics 67156 Frame integrator which will integrate 7 frames. I would like the capability to select more integration.
I have made a half-hearted effort to contact vendors on this matter, but I am not confident that I uncovered the full range of solutions.
In Activity 8 students examine sand and compare color, size, and shape of sand grains. They also look for crushed rocks, shells, bones, minerals, etc. Then they locate the sand on a map, thus linking science and geography.
Joe Neilly Abbott Laboratories
jfoust-at-threedee.com on 02/06/2001 09:58:29 AM To: microscopy-at-sparc5.microscopy.com, joe.p.neilly-at-abbott.com cc:
At 08:25 AM 2/5/01 -0600, Joseph Neilly wrote: } Past sand donations from listserver subscribers helped start the MSA sand } collection. This collection is freely given to educators to use with } Microscopic Explorations and other educational programs.
What exactly happens in Activity 8 of Microscopic Explorations? Are you looking for sand with special characteristics?
I would prefer not to take sides in the Darley/Garber discussion concerning Pt/C evaporation, but our research over the years clearly shows that it is the simultaneous nature of Pt/C evaporation which results in the finer grain structure. To achieve this we have long supplied Pt/C pellets (50/50) which as Dr. Danev described are placed between two flat faced carbon rods. If further information is required feel free to contact us or visit our web page.
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955
Jim at ProSciTech wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Chuck's reply without the customary inclusion of the previous contribution is } difficult to understand. I have now added my previous contribution and the } original request below. } The hanging drop method results largely in Pt evaporation followed by carbon. } The method also requires breaking the vacuum and is therefore more trouble. As } Sergey Ryazantsev in his thoughtful contribution explained (I erased that } because there was no need to reply), its the simultaneous nature of Pt/C which } yields the finer grain structure. Sergey also explained that the other means of } achieving such fine grain is electron beam evaporation, but that requires } special equipment. } I explained in my previous contribution why the thinner wire is required. } 30 years ago I used many meters annually of the 0.1mm Pt for freeze etching and } Kleinschmitt/ DNA rotary shadowing. } Nothing for it Chuck: you will need to stock 0.1mm Pt wire like all other major } suppliers. } Don't be overly concerned, very few people use these techniques now; in fact } you could purchase your minor requirements from us! } Disclaimer: its too obvious, we sell small quantities of 0.1mm Pt. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Sunday, February 04, 2001 12:43 AM, Garber, Charles A. } [SMTP:cgarber-at-2spi.com] wrote: } } ------------------------------------------------------------------------} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------.} } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } The "main" way Pt/C I thought was being evaporated today is the "bead on } } carbon" method, that is, where the wire is coiled up, around a pin of } } diameter slightly larger than the diameter of the neck on the carbon rod, } } and then, with some help from some good Style #5 tweezers, it is pulled onto } } the sharpened neck of the carbon rod. The bell jar is lowered over the } } carbon rods and holders, and wearing the appropriate eye protection, the } } power is slowly turned up, slowly heating the carbon rods and wound Pt wire. } } At some point the miracle happens: The Pt melts, and surface tension } } effects cause it to form (in an instant) a nice little "bead" (it looks like } } the textbook "sessile drop") firmly attached to the carbon rod once the } } heating is stopped. After cooling down, the rod with the drop is rotated } } around so that it is facing where the samples will be, the bell jar is then } } loaded, pumped down and evaporation of Pt/C can be done simultaneously this } } way. } } } } Indeed I don't think it is possible to evaporate Pt wire **without** the } } formation of the sessile drop, therefore, how the wire is originally spread } } out (on the rod neck) does not matter! } } } } A note not related to the original question: The sharpened "neck" should be } } sharpened to a diameter not more than 3/16" (4.75 mm). The method will not } } work as well if the neck diameter is larger than this. If you are } } sharpening your own rods, make sure you are using rods of higher rather than } } lower density, otherwise the rod will not have the mechanical properties } } needed to sharpen down to this diameter. } } } } I had a phone call on Friday from a customer asking about 0.1 mm (e.g. 4 mil } } ) wire since a previous posting seemed to attribute to it something special } } (relative to 0.2 mm/8 mil) wire. I told the caller that we did not believe } } that to be so, that for the practice of the sessile drop method, either } } diameter wire would "bead up" just as quickly and easily, and the only } } difference between the two diameters would be one of cost, since the cost to } } draw the same mass in 4 ml is obviously going to be higher than that to draw } } 8 mil. Actually some years ago we came into some 10 mil Pt wire, and we } } found it could be used just as easily to make the sessile drop. } } } } Furthermore since either diameter wire could be used to create a drop of } } equal size, the actual evaporation time would be independent of the diameter } } of the starting wire. } } } } In the end, we came upon the novel conclusion that I would make a posting } } and subject my advice to the most stringent (microscopy) peer review panel } } in the world, namely the listserver. Am I not correct, in that the end } } result will be the same irrespective of whether 4 mil or 8 mil wire is used? } } } } Chuck } } } } Disclaimer: SPI Supplies is a long time supplier of the high purity Pt wire } } that is used for evaporation in electron microscopy applications. } } } } =================================================== } } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } } President 1-(800)-2424-SPI } } SPI SUPPLIES FAX: 1-(610)-436-5755 } } PO BOX 656 e-mail: cgarber-at-2spi.com } } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } } } } Look for us! } } ############################ } } WWW: http://www.2spi.com } } ############################ } } ================================================== } } } From: Jim at ProSciTech [SMTP:jim-at-proscitech.com] } Sent: Friday, February 02, 2001 9:59 AM } To: 'Heinrich Matthies'; Microscopy-at-sparc5.microscopy.com } Subject: RE: rotary shadowing, platinum coating } } } I translate (x25.4) the funny measure 0.008 " to 0.2mm. } For Pt/C coating the more commonly used thickness is 0.1mm wire. If you work } out the cross section area of both wires you would find that you are using } considerably more Pt than the 50 to 75mm of 0.1mm wire otherwise used. } I suggest that you switch to the thinner wire because this would cover more of } the 1.4mm carbon cylinder. Therefore this would take longer (in time) to } evaporate. This is important because you need more than 5 seconds of } evaporation to obtain an even coating - with the specimen rotating at perhaps } 30 RPM. } What do you mean: " } at lower amps, less than 24, I don't see any Pt on the } test paper"? } The C would be much more visible than the Pt and normally you would not see the } Pt on filter paper. } The test is in the looking. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Thursday, February 01, 2001 9:00 AM, Heinrich Matthies } [SMTP:hjmatthies-at-ucdavis.edu] wrote: } } } } } } Hello, } } } } I'm plan to attempt to rotary shadow a motor protein using platinum on a } } carbon rod and then coat with carbon. At the moment, I am trying to } } evaporate the platinum from the carbon electrode and am having difficulties. } } } } We have a denton vacuum LLC with a bench top turbo III high vacuum } } evaporator. We wind one inch of platinum (Pt) wire on a nail (0.045) and } } then we load this 1 inch of 0.008 Pt wire onto a 0.04 tip of a 1.5mm carbon } } rod. The wire is pushed toward the solid carbon rod coming from the other } } side and all of the loops are tight (touch each other). The "spring" of Pt } } wire is tightly wound around the carbon wire. We pull a vacuum to about 5 } } x 10-5 torr and bring the amperage to 10 amps (filament 2) wait and then } } increase the amperage. Then I've tried 20,22.24, 26 and 30 amps but } } usually somewhere between 26-30, the carbon appears to evaporate. At lower } } amps, less than 24, I don't see any Pt on the test paper. So at the higher } } amps, both carbon and pt evaporate and this means we are getting too much } } carbon rather than an initial Pt coating. Under these conditions, the Pt } } wire only covers a short distance of the narrow tip of the carbon } } electrode. Does anyone have any advice or experienced this problem? } } } } Thank you, } } } } Heiner Matthies } } UC Davis } } MCB } } 530-754-9051
The message below, from John Francis of Polaroid Corporation, refers to a "Shutter Opening Utility" for cleaning dust off of the anti-aliasing filter of the Polaroid DMC digital camera. I tried it on one DMCie camera and the method was about 98% successful for removing dust particles between the shutter and filter. It took 1-2 hours to reach a point where I felt like it was as clean as I could get it. If the other 3 cameras can be cleaned this way, it will save us considerable time and money.
Gene Young
-----Original Message----- } From: Francis, John W [mailto:FRANCIJ-at-polaroid.com] Sent: Wednesday, February 07, 2001 8:01 AM To: 'Young, Gene (GP)'
Thanks to all who offered suggestions about doing an "antibody substraction." I shall proceed posthaste!
Jodi Schwarz phone: 541-737-4358 Zoology Department email: schwarzj-at-bcc.orst.edu 3029 Cordley Hall Oregon State University Corvallis, OR 97331 _______________________________________________________________
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} Email: kmccleary54-at-hotmail.com } Name: Kevin McCleary } School: South Peace Secondary School } State: British Columbia, Canada } } Question: I am anticipating the purchase of prepared slides for an } introduction to cells in my grade ten science classes.Ý Will chromosomes be } easily viewed in smears of Drosophila (advertized as giant chromosomes)? } If not, could you suggest alternate subjects.Ý Our light microscopes are of } standard high school quality. } _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
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MIDWEST MICROSCOPY AND MICROANALYSIS SOCIETY, INC. (MMMS) AFFILIATE OF THE MICROSCOPY SOCIETY OF AMERICA
MEETING ANNOUNCEMENT
JOINT MEETING WITH THE SOCIETY FOR APPLIED SPECTROSCOPY Tuesday, Feb. 13, 2001
At Unilever HPC USA, located at 3100 Golf Road, Rolling Meadows, IL. See directions for more details.
Social Hour: 5:30 pm Dinner: 6:00 pm Speaker: 7:00 pm _________________________________________________________________ The Infrared Microprobe in Production and Research By Koichi Nishikida
Abstract Unlike the visible microscope and electron microscope, which respectively use visible light and electrons to magnify the sample, the infrared microscope does not utilize infrared wavelengths to magnify the sample image. Instead, a visible microscope is modified, so that a magnified sample image is observed and, at the same time, an infrared beam is passed through the microscope to obtain an infrared spectrum of the sample. Therefore, the "IR microscope" should be better named the "IR microprobe".
Applications of the IR-microprobe cover applications from forensic } analysis to defect analysis, reverse engineering in the production industry, and medical diagnostic research.
In this talk, I will show how the IR-microprobe has contributed to quality improvements in the production industry, as well as recent research in medical applications. Briefly, I will address the evolution of this technique over the past five decades. ********************************************************************* Please make your dinner reservations for the upcoming meeting by calling (847)734-3712. Leave your name, company affiliation and the number of reservations. Please call by noon on February 9th, so that proper arrangements can be made. Dinner Cost: M3S Members: $25 M3S Students: $10 Nonmembers: $30
Directions to Unilever, Rolling Meadows, Illinois
From the Chicago Area: Kennedy Expressway to I-90 West (Rockford); I-90 West to Arlington Heights Rd.; North to Golf Rd. (2nd traffic light); Turn left (West) onto Golf Rd. for approximately 2 miles. The Unilever R&D facility is on the right, just before Hwy. 53, across from the forest preserve. Follow the entrance road all the way to the last parking lot (North lot) and enter the building through the far glass doors.
From I-290 Eisenhower Expressway: Stay on I-290 towards Rockford; Exit on Golf/Higgins Rds.; Continue North to Golf Rd.; Turn east on Golf Rd. Unilever is1/4 mile left on Golf Rd., across from the forest preserve. Follow the entrance road all the way to the last parking lot (North lot) and enter the building through the far glass doors.
From the Northern Suburbs (Route 53 South): Exit Woodfield Drive/Golf Rd. (58); Continue to first light; Turn left at first light; Continue to Golf Rd (58); Turn right (East) on Golf Rd.; Unilever is1/4 mile left on Golf Rd., across from the forest preserve. Follow the entrance road all the way to the last parking lot (North lot) and enter the building through the far glass doors.
If you have any questions about the meeting, please direct your inquiries to the phone number below. For questions about MMMS, please contact:
Dr. Damian Neuberger Senior Research Scientist Baxter Healthcare, Corp. (847) 270-5888 damian_neuberger-at-baxter.com
I suppose that this is nice, but one must wonder why such kludge operations are necessary in the first place? It seems to me that it is simply indicative of a basic flaw or limitation in the unit's design. The DMC is rather old, in technology life time terms. It was rather good in its day, but that has passed.
If this operation does keep your investment working, that is good. If the periodic procedure does not become too burdensome, then the unit should serve one's needs.
I had one of these units for about 4 months and found that it not only did not perform well but was quite unreliable. Polaroid was very accommodating in offering a refund in lieu of a replacement. Their scanners are quite the opposite of their cameras, in my experience.
gg
At 02:27 PM 2/7/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi, has anyone done any study or comparison of the slow scan CCD camera from Gatan and Tietz, for example, Model 795 from Gatan vs. Model TemCam-F224 from Tietz? Any comments will be appreciated.
Sam Li ******************************* Medical Research Council Laboratory of Molecular Biology Hills Road Cambridge, CB2 2QH United Kingdom *******************************
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Brian,
there are definitely many cards out there to acquire standard TV signals from any source that adheres to that standard. These standards were developed decades ago for TV cameras, and my personal opinion is, that they are good for moving images (as on TV), but they lack resolution and definition for still images (LM and SEM). However, since they cards are usually not very expensive, and some of them do offer integration capabilities (normally frame integration), it might be worth a shot. Also the microscopes themselves might be offering integration capabilities. You are limited to a resolution of 640x480 (NTSC) or 758x576 (PAL).
For better resolution you might want to look at other options as well. For SEMs there are interfaces available (such as our ADDA II, but there are other manufacturers also), normally available as "passive" or "active" systems. For LM, of course, there are digital cameras with better resolution and bit-depth (Video only carries about 8 bits of information, if you're lucky).
Look for systems that are open for upward expansion to keep your options open. Contact me offline if you are interested in getting more information about the way we do that.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
At 07:03 AM 1/29/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a Sorvall MT 6000 that sections much slower than the millimeter/second speed I have it set at. I can get it to operate correctly by speeding up the set speed and letting it run at that setting for awhile and then slowly taking it down to my desired sectioning speed. I have cleaned it, but don't know what else to do. Any suggestiongs?
Thanks.
Phoebe
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Department of Physiological Sciences 264 McElroy Hall Oklahoma State University Stillwater, OK 74078 405-744-6765
We have an MT5000 which works though is surplus to our needs, I am happy to ship it to the most needy home Simon
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 http://sbic6.sbic.pitt.edu -----------------------------------
I am looking for B/W CCD cameras that, mounted on a microscope, can be connected and directly controlled (not driven from an extensive package, only by a simple driver) by a Silicon Graphics O2 workstation. The chip should at least have 1030x1030 (about 6 micron square) pixels. It should have 8 to 12 bit greyvalues. It will NOT be used for live images (25 full frames in Europe), so (transmission) speed is not an issue. It should have exposure times between 10 ms and a few seconds. The camera has to be controlled (setting exposure time and taking an image) by the SGI-O2. If anyone has information I hope you will be so kind as to share it with me. Please respond directly to me (email address below). Thank you all in advance.
Kees van der Wulp
C.J.M. van der Wulp TNO-Prins Maurits Laboratorium Division 3, dept. MB Lange Kleiweg 137 2288 GJ Rijswijk The Netherlands
Am looking fo a sem operator/metallurgist/materials person for a small optical and electron microscopy lab in the Detroit area. If interested call 734-668-3309 or e-mail Dick O'Connell at oconnell-at-LTU.edu
Does anyone know what Amplicons (from HSV I) look like either in cultured cells or by negative staining. I have done several literature searches which discuss the molecular structure with diagrams, but none of them have a TEM image.
Karen L. Jensen, M.S. Associate Scientist & Project Manager Electron Microscope Research Core Pathology Department, Box 626 University of Rochester Medical Center Rochester, NY 14642
I feel like an E-Bay auction (though this has no $$s attached)... Thanks for the tremendous interest (we received almost 50 requests). The MT5000 is on its way to Oshkosh Wisconsin, to be placed in the lab of a starting faculty member who is putting a new EM facility into operation. Simon
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 http://sbic6.sbic.pitt.edu -----------------------------------
We have a Gatan Dual Ion Mill model 600, which we no longer need. It is complete and working (as far as we know) except that it lacks a backing pump.
Will anyone who is interested in acquiring it please contact me (off line, of course). .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
*************************************************** FOCUS ON MICROSCOPY 2001, Amsterdam, April 1-4 ***************************************************
We are happy to announce that about 100 people have submitted an abstract. Although the deadline for abstract submission was a week ago (February 1), we still receive new abstracts every day. Next week we will finalize the program and put the final program on the web on February 19. So, if you would like to submit you contribution for FOM 2001, please hurry!!!
On our web-pages you will find the titles of some of the presentations of the invited speakers. With the 100 contribution from other researchers from various fields (e.g. instrumentation, non-linear optics, 2-photon microscopy, imaging of living cells), the conference will become a great success.
We expect that about 250 participants will visit the conference. Since the maximum number of participants is 300, it is important to register early !!!
*************************************************** FOCUS ON MICROSCOPY 2001, Register now !!!! ***************************************************
FOM 2001 , Amsterdam The Netherlands, April 1-4, 2001
FOM 2001 is a world leading international conference on advanced microscopy. It is the joint meeting of the 13th International Conference on Confocal Microscopy and the 13th International Conference on 3D Image Processing in Microscopy.
FOM 2001 will be held at the Academic Medical Centre (AMC) of the University of Amsterdam, Amsterdam, The Netherlands.
For more information visit: http://www.focusonmicroscopy.org
Core conference subjects: "INSTRUMENTATION" and "IMAGE PROCESSING AND ANALYSIS"
SPEAKERS: S. Hell (Gottingen, Germany), T. Wilson (Oxford, UK), S. Kawata (Osaka, Japan) C.J.R. Sheppard (Sydney, Astralia), C. Coggswell (Sydney, Astralia) E. Stelzer (Heidelberg, Germany) C. Cremer (Heidelberg, Germany), P.A. Benedetti (Pisa, Italy), A. Kriete (Giessen, Germany), H. van der Voort (Hilversum, The Netherlands), P.C. Cheng (Buffalo, USA), G.J. Brakenhoff (Amsterdam, The Netherlands)
Special conference subjects this year: MULTI-PHOTON MICROSCOPY" and "MICROSCOPY OF LIVING CELLS AND TISSUE".
SPEAKERS: A. Zumbusch (Muenchen, Germany), L. Moreaux (Paris, France) A. Diaspro (Genova, Italy), M. Muller (Amsterdam, The Netherlands), K. Sullivan (La Jolla, USA, to be confirmed), R. van Driel (Amsterdam, The Netherlands), R. Eils (Heidelberg, Germany), T. Mistelli (Bethesda, USA; to be confirmed), D. Gadella (Wageningen, The Netherlands), J. Dobrucki (Krakov, Poland), A. Houtsmuller (Rotterdam, The Netherlands), E. Manders (Amsterdam, The Netherlands)
To the Listserver I have an investigator who needs some EDS work done. My lab does not have a capability to do this type of work.
The investigator is looking for evidence of Aluminum Oxide deposition in six tissue samples of lung, brain, and skin obtained from mice exposed to and not exposed to aerosolized Aluminum Oxide. They are willing to send the samples to any lab in the United States who can do the work for them. Please email me if you can help.
Thanks in advance,
George
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 Fax 214-648-6408 eMail: George.Lawton-at-UTSouthwestern.edu
GR Newman introduced LR-White for Immuno-EM 1982/1983. The first article published was in Histochem J (1983,15,543-555). OK so far. But there was a earlier report in the Journal of Microscopy in abstract form. I know issue and pages from another cite (J Microscopy, 1982, p. 127, RP5-6), but I don´t know the TITLE ....! Who can help me? Tanks in advance, With best regards,
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
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I would like to request of preparation samples from poly(styrene)/poly(butyl acrylate) dispersion with 100 nm particles size for measurements by JEOL type of transmission electron microscope. We have got some problems with dilution of latexes and with time of staining of carbon, too. We would like to confirm a core/shell particles morphology of samples prepared by two step seeded emulsion polymerization. We have got some problems with magnification and contrast expansion of core/shell particles,too. Thank you very much for your help.
Petra Volfova, PhD. student of Department of plastics and rubber, Slovak Technical University,Bratislava,Slovak Republic.
NEWMAN GR, JASANI B, WILLIAMS ED THE PRESERVATION OF ULTRASTRUCTURE AND ANTIGENICITY J MICROSC-OXFORD 127: (SEP) RP5-RP6 1982
Chris
----- Original Message ----- } From: "Michael Reiner" {Elektronenmikroskopie-at-web.de} To: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, February 10, 2001 12:13 PM
Dear members of the listserver,
GR Newman introduced LR-White for Immuno-EM 1982/1983. The first article published was in Histochem J (1983,15,543-555). OK so far. But there was a earlier report in the Journal of Microscopy in abstract form. I know issue and pages from another cite (J Microscopy, 1982, p. 127, RP5-6), but I don´t know the TITLE ....! Who can help me? Tanks in advance, With best regards,
Michael Reiner University of Cologne, Germany Dept. of Anatomy I ______________________________________________________________________ _________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
I did some experiments with my SEM and external frame grabber. The results were rather interesting.
SEM is an Amray 1910FE, which includes internal TV signal processing, which is frame averaging and Kalman filtering.
External frame grabber is a Soft Imaging GrabBit PCI board controlled by their analySIS software. The analySIS and GrabBit package will do a huge number of frame averages. It will not do Kalman.
If I do frame averaging in the SEM at a count of 4 or 5, I get rather good results. If I do Kalman, I get very good results. Either of these frame processes can be frozen in the SEM's internal buffer. That composite NTSC RS-190 video is grabbed by the GrabBit. The captured results are those seen on the SEM's monitor.
If I do frame averaging with the GrabBit and analySIS anywhere from 8-30 frames, the results are not near as good as using the SEM's electronics. At low to medium magnification ( { 10KX), running at 80 averaged frames makes no dramatic difference. As magnification increases, the image tends to slightly shift. This effectively kills the ability to average. However, the SEM's internal Kalman filter will capture a very nice image in about 2-3 seconds. And this is quick enough to avoid image movement.
So, to answer your question, I would suggest looking for a TV frame grabbing system which does Kalman. No idea if such a thing exists. It would probably be an integrated h/w + s/w product since they would have to be closely tied. Since it is a real time process, the software would have to process image frames directly from the frame grabber board/interface.
If you need any other info, please contact me.
gary g.
At 02:41 AM 2/7/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Question: I wanted to know whether there are any books available which issues the topic of SEM analysis of optical fibers, which helps in analyzing the SEM images of optical fibers for Break source Analysis. Also I would like to know whether there are any other materials on this subject
} } } Hi, } } I am trying to identify an onset of apoptosis in endothelial cells (brain } } tissue) using postembedding immunostaining method. So far I have tried 11 } } different AB's and have not } } been able to stain the cells I know should be staining. Tissue has } } already been embedded in lowacryl so i have no choice in the run up. Any } } suggestions???? Neelima Shah.............. Biomedical Imaging Core Facility Uni of Pennsylvania Philadelphia, Pa.
Many years ago, probably during the mid '60's, I saw an instrument that was described to me as a "Beta Probe". It was used in the lab of a company selling non-ferrous metals (principally brasses and bronzes), and was an analytical instrument which determined the composition of the products with enough precision to qualify them for government specifications.
I remember parts of the demonstration I was given. A chunk of the material was ground and polished, then simply set on an O-ring seal on the top of the device. It was evacuated, the "Go" button was pushed, and after a suitable time (and perhaps other manipulation) some counters were read. Presumably these numbers were then converted, via a calibration, to the composition.
At the time I had no clue what it was actually doing. Now I surmise that it was an electron probe (probably with a "macro" spot size) with pre-set crystal spectrometers.
If anyone knows anything more about this device, both I, and my aunt (who managed the chemical part of the laboratory at that time - hence my visit!) would be fascinated to know. Many thanks.
I am member of few entomological listserver, even I am owner and moderator of coleoptera listserver. What I do not understand why only this listserver has spam messages. But When I try to write any message I am rejected..
Polarized Light Microscopy April 21, 28, May 5, 12, 2001
An advanced course on polarized light microscopy which will cover the following topics: The nature of polarized light The origin and interpretation of interference colors Birefringence and crystal orientation The Indicatrix Compensation and variable compensators Interference figures and their interpretation
The workshop will consist of four consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy. The course instructors include Jan Hinsch of Leica, Inc., Mary McCann of McCann Imaging, John Reffner of Sensir Technologies and N.Y.M.S. Instructor Don O'Leary.
WHEN: April 21, 28, May 5, 12, 2001 from 10 A.M. to 4 P.M.
WHERE: “Evergreens”, 30 N. Mountain Avenue, Montclair, NJ 07042 (accessible by public transportation, Information on car pools and transportation will be provided.)
COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants. Return form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849
PLEASE POST ---------------------------------------------------------------------------- ------------------------------------- Registration Form Polarized Light Microscopy
One of Murphy's laws says that whenever something is published, it's immediately out of date. Project MICRO's "Microscopy for Children: a Bibliography", which was distributed with the December "Microscopy Today", is no exception. I've been contacted recently about two CD-ROMs that will interest many listserver readers. Since the web version of the bibliography (URL below) won't be updated for a while, I'm providing listings here:
Edmark Corp. 1998 Zap! $29.95, with a 50 page users' manual, from Edmark at PO Box 97201, Redmond, WA 98073-9721, 800-320-8379, www.edmark.com. For Macintosh or Windows 3.1 or 95. Subtitled "Save the show with sound, light, and electricity", this CD-ROM is sure to capture the interest of a computer game-addicted middle schooler. All three topics are introduced in game format, complete with levels of complexity and lots of shooting. In the optics section, laser beams are aimed at mirrors, lenses, prisms, and filters; the targets are eggs that hatch into cute monsters when activated by the beam. It actually does a very good job of teaching reflection, refraction, absorption, and color. The player who works through all levels of all three units then can set up the light show for a concert, and even manipulate the sound. It's much too time consuming to use in the classroom, but it's a delight for home use. There is ample reference material, and homeschool parents can set options to control complexity. Middle school. RECOMMENDED
Excalibur Mineral Company 1999 Photographic Guide to Mineral Species $69.95+ $4.00 shipping from Excalibur at 1000 North Division SStreet, Peekskill, NY 10566, www.bestweb.net/~excalmin. For Windows or Mac; requires Netscape Communicator (included). This is an "adult" CD, but the superb photos make it an excellent visual supplement for a student who is learning about crystals and other minerals. It's truly a "coffee table book" on a CD, with an unbelievable 5400 photos, which search and load rapidly in Netscape (and not at all in Internet Explorer). There is an eye-catching 300 image automatic slide show which can be set to play continuously. Images can be searched by location or mineral content. They are mostly micromounts, photographed at dissecting scope magnifications. Descriptions are minimal; it isn't intended as a textbook substitute. Adult. RECOMMENDED
Want more? There are over 20 on the website. Want copies of the bibliography for educational use? Contact me.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Dear Neelima, wow, 11 different antibodies tested is a lot! I don´t think that your problem is the choice of the antibody, you are enfaced with a more general problem.
Let´s assume your sections are about 60 nm thick. This is about a 100 times thinner than a conventional paraffin section and about 200 times thinner than a conventional cryosection. Thus, you have to consider that the amount of accesible epitopes/signal is 100-200 times weaker as seen in LM. Then, your samples are already embedded in resin. I don´t know what kind of procedure you applied, but consider a factor } = 10 of epitopes destroyed during embedding. That means at least 1000 times less epitopes in the same visible area as in LM-IHC. Impressive.
Now the other way round: Let´s say a section of 1mm in square of (mouse) brain cortex contains about 50 capillaries and 3-4 greater vessels, the capillaries containing 2-3 endothelial cells hit. That makes altogether, mh, 200 endothelial cells in your section (not whole cells, only a 60 nm slice of them...). I don´t know if you are examing normal tissue or pathological alterated (hypoxic, transgenic, inflamed, radiated etc...) but generally the apoptosis rate of adult continous endothelium is quite low. Let´s say rate of apoptosis is 2%, that would mean that 3-4 cells in your 1mm in square area are apoptotic! Common apoptosis markers refer to the nucleus. If 25% of all the endothelial cells show a nucleus hit by the section plane - expect one cell positive. Even if apoptosis rate is 10% there would be optimistically 5 cells, including those who are in the holes of your section after the immunoprocedure :-(
In my point of view, you are looking for the needle in the haystack.
If I had to look for apoptotic cells by immunohistochemistry, I would take thick sections of mildly fixed tissue on a vibratome and do a discrete DAB-reaction-based IHC after the use of a detergent (e.g. AB against p18 of PARP). Then embed the sections with positive cells after osmification and focus on trimming your blocks towards the cells positive. (a pity that you have no choice in the run-up)
In the proceeded apoptosis of the cell you can observe typical changes in the nucleus and cytoplasm, so the use of an antibody is not the only way to look for apoptosis at EM level.
I´m curious what the other people on the listserver have to say to this topic.
I wish you good luck and cross my fingers for you.
Best regards, Michael
Michael Reiner University of Cologne Dept. of Anatomy I Germany
Neelima Shah wrote: Hi, I am trying to identify an onset of apoptosis in endothelial cells (brain tissue) using postembedding immunostaining method. So far I have tried 11 different AB's and have not been able to stain the cells I know should be staining. Tissue has already been embedded in lowacryl so i have no choice in the run up. Any suggestions????
Neelima Shah Biomedical Imaging Core Facility Uni of Pennsylvania Philadelphia, Pa.
_______________________________________________________________________________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
Urgent request Is there anyone out there in cyber space that may know where we can obtain the metal band (LKB part number 90-00-0047) that hold the specimen arm up on an LKB Nova ultramicrotome. Any help would be gratefully appreciated
Terry
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6009
Urgent request Is there anyone out there in cyber space that may know where we can obtain the metal band (LKB part number 90-00-0047) that hold the specimen arm up on an LKB Nova ultramicrotome. Any help would be gratefully appreciated
Terry
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6009
I would like to purchase the Photoshop image software package. Unfortunately our computer department does not have any information on the cost of this package or where it can be purchased. Who can I contact for this information ?
Regards W. Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9603954 Fax : +27 +16 9602826 E-mail : willem.erasmus-at-sasol.com PO Box 1, Sasolburg, 1947, Republic of South Africa
[All views expressed are my own and not necessarily that of my employer.]
Leica now handle LKB/Reichert microtome parts if you can still get them. I am trying to trace a part in the UK for myself so I will contact you if they come back with any information on your metal band, it may take two weeks though. You could however try contacting Leica Australia see web address for details: Main web page http://www.leica-microsystems.com/ Australian website http://www.leica-microsystems.com/cgi-bin/boxgate29910?login=guest&password=&opt=area%3DCompany
I get the impression that the normal electron microscope rule of at least 10 (maybe 15 if you're lucky) years of availability for spare parts won't apply to ultramicrotomes because of the major changes in the market about 12 or 13 years ago (in the UK: LKB ---} Reichert ---} Cambridge Instruments? ---} Leica).
Good luck
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
Terry Robertson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Urgent request } Is there anyone out there in cyber space that may know where we can obtain the metal band (LKB part number 90-00-0047) that hold the specimen arm up on an LKB Nova ultramicrotome. Any help would be gratefully appreciated } } Terry } } Dr Terry Robertson (PhD) } Senior Research Fellow } Department of Pathology } University of Western Australia } Nedlands 6009 } } Phone 618 9346 2935 } Fax 618 9346 2891 } Mobile phone 040302 5440 } email terryr-at-cyllene.uwa.edu.au
2001 MICROSCOPY TECHNICIAN SUMMER POSITION AVAILABLE
The Marine Biological Laboratory has a summer position available for 10 to 15 weeks (June, July, and August) for a microscopy oriented technician. We would like to attract someone with some knowledge of biological preparative techniques and experience in any of the following: laser scanning confocal microscopy, TEM, SEM, and/or LM.
The technician will assist in the Central Microscopy Facility. The technician's duties will be to check out incoming investigators in the usage of our equipment and then to supervise its continuing usage and to perform contract work for investigators. This may include fixation, embedding, sectioning, scope use, darkroom work, etc. The technician will also provide routine maintenance.
This is a short term and scientifically rewarding position. Salary will be in the $8 to $10/hour range. Housing may be available to rent through MBL.
For more information, including a more detailed position description, please contact Louis Kerr at: MBL, 7 MBL Street, Woods Hole, MA 02543. Telephone, 508-289-7273; or at Email: lkerr-at-mbl.edu.
Please apply to: Human Resources, MBL, 7 MBL Street, Woods Hole, MA 02543. or resume-at-mbl.edu.
You can visit our web site for online employment information and general information regarding the Marine Biological Laboratory - http://www.mbl.edu/
An Equal Opportunity/Affirmative Action Employer/ Non-smoking workplace.
I'm looking for a service (in the USA) where I can send my TEM blocks to be sectioned and stained (UA/LC). These are biological samples cured in Embed 812 resin.
Lawrence Mason Research Scientist Genetics Institute lmason-at-genetics.com
Mayo Clinic has an opening for an EM technician in the Muscle Research Lab at the Rochester, MN campus. Details are as follows:
Research Technologist ñ Posting #01-0818
Summary:
Responsible for preparation of specimens for electron microscopy, evaluation of specimens in electron microscope, and related photography work. Will participate in research projects involving transmission EM, EM cytochemistry, and immunocytochemistry; data analysis including morphometry; assist in preparation of manuscripts for publication. Innovative and creative individual capable of independent work with minimal supervision is preferred. Executes experimental studies in support of lab goals, grant commitments. Assists in the planning, design and modifications of experiments. Performs basic statistical data analysis; prepares tables/charts/graphs and assists in organizing data. May make presentations and may contribute to writing of manuscripts. Trains residents/fellows/temporary lab personnel in lab techniques. Maintains lab equipment and supplies.
Qualifications:
Requires a Bachelor's degree in biology, chemistry, or other relevant sciences. Experience required in lab projects/research work in educational or work setting which would provide a solid understanding of lab techniques, equipment, and safety. Must have the ability to organize and carry out basic techniques independently. Candidate should have the ability to work both independently and in a team. Experience with electron microscopy is preferred.
For additional information on this position refer to posting # 01-0818 and contact:
Jill M. Kelly Staffing Specialist Mayo Clinic - Ozmun East -4 kelly.jill-at-mayo.edu ph #(507) 284-0510 fax# (507)284-9751 Assistant: Leah Carlson (507)284-3574
I am very interested in getting softwares to process electron diffraction patterns from disordered materials to obtain the G(r) (reduced intensity function), and J(r) (radial distribution function).
Any information on how to get these softwares is appreciated. In particular, information on free ones are most welcome.
Regards Yan Xin
======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
Qualified candidates (Ph.D. or other terminal degree) are sought for a NIH-funded postdoctoral position to study intracellular calcium signals and their downstream effectors which regulate myofibril assembly. Primary training in real-time fluorescence digital microscopy (both conventional and confocal) using calcium tools and fluorescently tagged sarcomeric proteins expressed or microinjected in cultured myocytes. Prior training in protein biochemistry or molecular biology highly desirable. Interests and skill areas could include: cell signaling, cytoskeleton, actin or myosin dynamics, intracellular calcium channels, muscle development. Salary and start date are negotiable, funding available summer 2001. Please reply offline. See website below for more info.
Mike
Michael B. Ferrari Department of Biological Sciences University of Arkansas Fayetteville, Arkansas 72701 Ph: 501-575-6372 Fax: 501-575-5349 http://biology.uark.edu/ferrari
Has anyone out there in immuno cyber space had any experience in demonstrating complement C3 or C5 on formalin fixed frozen or paraffin sections. I need a good antibody that works. Any comments would be gratefully received.
Terry
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6009
Someone from another lab that used to do EM, but doesn't do EM any more just delivered a couple of boxes of old resin components to me to 'take care of'.
It was OK with me, being the EM guy I should know what to do with them, but I realized I wasn't sure. I already had a bunch of old resins left over from kits folks had bought through the years and I figured I could just add it to the pile, but now there is a space problem, too may old bottles and not enough space to keep them.
These things are old, 1980's, but could they still be good enough to use? Is it worth finding space for them in case some poor starving student comes along who can't afford new supplies but is dying to do EM? What is the shelf life of things like this?
What would you do? Mix up a bunch of paper weights? Call EH&S and have them take away the bottles of individual components? Consolidate the nearly empty bottles to save space? Would you ever use things this old, I don't think I would. I am tempted to just get rid of them in one way or another. But I am haunted by the possibility that I will be tossing perfectly good stuff.
Relieve me of my guilt, tell me what to do.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Don't waste your time, disgard the lot. Regards JVN
Jon Krupp wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi: } } Someone from another lab that used to do EM, but doesn't do EM any more } just delivered a couple of boxes of old resin components to me to 'take } care of'. } } It was OK with me, being the EM guy I should know what to do with them, but } I realized I wasn't sure. I already had a bunch of old resins left over } from kits folks had bought through the years and I figured I could just add } it to the pile, but now there is a space problem, too may old bottles and } not enough space to keep them. } } These things are old, 1980's, but could they still be good enough to use? } Is it worth finding space for them in case some poor starving student comes } along who can't afford new supplies but is dying to do EM? What is the } shelf life of things like this? } } What would you do? Mix up a bunch of paper weights? Call EH&S and have them } take away the bottles of individual components? Consolidate the nearly } empty bottles to save space? Would you ever use things this old, I don't } think I would. I am tempted to just get rid of them in one way or another. } But I am haunted by the possibility that I will be tossing perfectly good } stuff. } } Relieve me of my guilt, tell me what to do. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
Job Hunters, There was a job offer on the server last month from RangeTS-at-aol.com for an SEM tech job in Florida. Did anyone send any info and, if so, did anyone get a reply? Concerns have been expressed to me about having sent off personal info to a blind add and then getting no reply.
If RangeTS-at-aol.com is lurking out there, please have the courtesy to reply to all who answered your add ASAP. Taking personal info without acknowledgement or info on who you are is rather unprofessional. Blind adds are not the best way to go, but if you feel you must use them, the good folks on this listserver expect a professional response. Thank you.
Ken Converse owner Quality Images third party SEM service Delta, PA 17314
I am interested in characterizing emulsion latex Pressure Sensitive Adhesives (PSA) in their aqueous phase state and as cast films. Can anyone offer any specific techniques tips for doing cryo- ultramicrotomy of such material? Are there any good literature references that would be informative? Any recomendations for staining techniques of Acrylic based copolymer based adhesives?
At 04:38 PM 2/12/01 -0800, Jon Krupp wrote: } Someone from another lab that used to do EM, but doesn't do EM any more } just delivered a couple of boxes of old resin components to me to 'take } care of'.
I wholeheartedly encourage this kind of behavior... Tell the mailing list about old stuff you're about to throw away!
Someone out there might be willing to pay the cost of shipping and handling to get it, and perhaps they'll put it to good use.
It may not need to be fresh, contemporary or professional quality; don't forget the schools, teachers, amateurs, etc.
Hello All, Can anyone suggest/recommend a fixation protocol that preserves microtubules and/or centrioles without needlessly extracting other cellular components?
Thanks for any response, Jay Campbell University of Wisconsin Microscopy Resource
I sent some of the requested info, but I held some back just in case the request was bogus. It seems my fears are justified, as I never received a reply. On the other hand, I have not received any new spam, so perhaps it's not a diabolical plot. ;-)
I do hope, if it was a legitimate offer, that RangeTS-at-aol.com will take the time to respond to those who expressed an interest.
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Ken Converse {qualityimages-at-n To: "MSA, listserver" {Microscopy-at-sparc5.microscopy.com} etrax.net} cc: Subject: Re:job offer from RangeTS-at-aol.com 02/13/01 06:47 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Job Hunters, There was a job offer on the server last month from RangeTS-at-aol.com for an SEM tech job in Florida. Did anyone send any info and, if so, did anyone get a reply? Concerns have been expressed to me about having sent off personal info to a blind add and then getting no reply.
If RangeTS-at-aol.com is lurking out there, please have the courtesy to reply to all who answered your add ASAP. Taking personal info without acknowledgement or info on who you are is rather unprofessional. Blind adds are not the best way to go, but if you feel you must use them, the good folks on this listserver expect a professional response. Thank you.
Ken Converse owner Quality Images third party SEM service Delta, PA 17314
Warren wrote the following: ======================================================= I am interested in characterizing emulsion latex Pressure Sensitive Adhesives (PSA) in their aqueous phase state and as cast films. Can anyone offer any specific techniques tips for doing cryo- ultramicrotomy of such material? Are there any good literature references that would be informative? Any recomendations for staining techniques of Acrylic based copolymer based adhesives? ======================================================= This is not a question that is easily answered because you have to state further just what are your analytical objectives for wanting to do this work in the first place.
But if a typical (some are not "typical") acrylic emulsion, then the "particles" have to be "hardened" up in some way, otherwise they will coalesce and pancake, and when viewed by TEM will, otherwise, be one big ambiguous mess. So the soft particles have to be hardened up (into what look like hard rigid "marbles") and that is generally done using UV irradiation in quartz tubes or sometimes using RuO4. We get the best results on these kinds of emulsions with samples lightly shadowed with Pt/C.
For the sectioning of the resulting adhesive film, again this is a TEM job, you have to define whether you want to look at the interface with the release paper or some other characteristics, such as the dispersion of inorganic additives in the adhesive coating. Evidence of the release layer can sometimes be resolved this way as well. The preparation methods are different (for the different objectives) but the common feature is that they all must be cryo-sectioning using a good state of the art cryo- ultramicrotome and using only good quality diamond knives. Anyone who has either tried or done this knows that it can take one quite a long time to develop the magic of the "art" before good sections can be made on a reproducible basis.
I would be happy to give you more information off-line or better yet, our laboratory services division would be very happy to give you a quote to do the work for you.
Disclaimer: Our firm manufactures and distributes the kinds of materials and supplies needed for the conduct of this type of work and when one prefers to send it to an outside laboratory, we also perform studies on these kinds of samples for clients in our own facilities.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} Hello All, } Can anyone suggest/recommend a fixation protocol that preserves } microtubules and/or centrioles without needlessly extracting other } cellular components? } } Thanks for any response, } Jay Campbell } University of Wisconsin } Microscopy Resource
You might have a look at "Increased visualization of microtubules by an improved fixation method". Luftig, R. B. et al. J. Histochem. Cytochem. 25:175-187, 1977. I seem to recall someone using PIPES as a buffer for glut fixation to improve microtubules.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Hi Jay, You didn't say what kind of cells so I won't comment specifically. But in general a good fix for tubes will have 10 mM EGTA to chelate calcium. A typical fix might contain 4 % paraformaldehyde, 0.1% glut, 25 mM Pipes pH 7.0, 10 mM EGTA, 1 mM MgSO4. Remember to fix at room temp, at least for the first half hour or so, because microtubules are cold sensitive. Hope this helps, Tobias
} } } } Hello All, } Can anyone suggest/recommend a fixation protocol that preserves } microtubules and/or centrioles without needlessly extracting other } cellular components? } } Thanks for any response, } Jay Campbell } University of Wisconsin } Microscopy Resource
I have extensive experience in all types of biological specimens and TEM. If you would like to discuss your project and estimated costs, please contact me at 716-275-1954. Our EM laboratory is housed in the Pathology Department, but serves as a core facility to any University of Rochester Med. Ctr. researchers. We also do projects for companies and researchers outside the University. So please call if you are interested in a discussion...
Sincerely,
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core, Box 626 University of Rochester Medical Center Rochester, NY 14642
I disagree with JVN, because if the bottles have never been opened then they can definitely be used. Other EM technicians I have surveyed here at the University of Rochester Medical Center in Rochester, NY have used resins purchased in the 1980's in the 1990's-2000. So, if they are resins that you routinely order, start using the older bottles. Save yourself some money! Also you can test embedd a specimen. By the way if any of the resins are Lowicryls, they are definitely OK unless they have turned color, should be white not pale yellow. Also, here at the U of R we have a chemical recycle list that is accessible to other researchers here, thereby providing an opportunity to use unopened resins, chemicals, etc...Maybe your university has a similar program.
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core
Petra Volfova wrote: } } I would like to request of preparation samples from } poly(styrene)/poly(butyl acrylate) dispersion with 100 nm particles size } for measurements by JEOL type of transmission electron microscope. } We have got some problems with dilution of latexes and with time of } staining of carbon, too. We would like to confirm a core/shell } particles morphology of samples prepared by two step seeded emulsion } polymerization. We have got some problems with magnification and contrast } expansion of core/shell particles,too. } Thank you very much for your help. } } Petra Volfova, PhD. student of Department of plastics and rubber, } Slovak Technical University,Bratislava,Slovak Republic. }
Dear Petra, Below are my experiences with TEM on polystyrene/poly(butyl acrylate) latexes. Please note that I'm not a specialist in this area and these are _not_ 100% perfect receipts - but it often works for me. Dilution: Wet the end (1...2cm) of a glass stick with your latex and dip it in a test tube filled with ca. 10 ml dist. water. The water must become slightly opaque. Repeat one or two times (depending from the concentration of the latex). Staining: If poly(butyl acrylate) forms the shell, some negativ staining could be helpful, because the acrylat phase itself is often not visible in TEM. I used uranyl acetate, two drops of 2% UA solution in 10 ml diluted latex. The PS-phase can be stained by vapour staining with Ruthenium (0.1g RuCl3 + 5 ml Na-Hypochlorid solution (13% Cl)). Preparation: I've mounted copper grids, coated with a 10 nm carbon film, on a specimen holder (from a TESLA BS500 TEM) and placed a drop diluted latex on the grid (formvar or collodium coated grids are not recommendable, because these films are very unstable under the electron beam after Ru-staining). After drying (some hours at room temp.) the grid can be umounted for vapour staining. If you have an "ultrasonic fogger" (or how this device is called in english, in german it's an "Ultraschallvernebler"), you can use it with very good results for preparation of latices smaller than ca. 200 nm. Microscopy: There is nothing special, if you have problems with the contrast, play a little bit with the accelerating voltage and objective aperture. Magnification should be not the problem. In polystyrene-core/poly(butyl acrylate)-shell particles the core/shell structure should be visible (if the acrylate shell is not too thin). If PS forms the shell, I've never seen a core/shell structure. In this case, you must embed the dryed latex particles in a resin (acrylate or epoxy), cut ultrathin sections and stain them for visualization of the core/shell structure (if the particles are not crosslinked, they may be soluble in some kind of resin, especially at higher temperatures).
Literature, that may help: A.W. Robards, A.J. Wilson (editors): Procedures in Electron Microscopy; John Wiley and Sons, 1993-1998 (a comprehensive collection of receipts, eg. staining methods, preparation of carbon and formvar coated grids etc.)
A technical position is available at the Eye Research Institute, Toronto Western Research Institute, University Health Network. Experience in immunohistochemistry and histology is essential, and a background in molecular biology would be beneficial. The position can either be a full time junior post, or part time senior post. The successful candidate will assist investigators who study the molecular mechanisms of eye development and disease. A CV and the names/phone numbers of two referees should be sent to Rod Bremner,
Judy Trogadis Vision Science Research Program Toronto Western Research Institute 399 Bathurst St. Toronto, ON M5T 2S8, Canada ph: 416-603-5088 fax: 416-603-5126 email:judy-at-uhnres.utoronto.ca
Dear Fellow Microscopists...I'm looking for suggestions on how to plan an undergraduate forensic microscopy course. Chances are the course will have to start with a generic introduction to microscopy, so anything on general undergraduate microscopy would also be appreciated. Books? Demonstrative Aids? Personal experiences with undergrad instruction, Etc. Any and all help would be appreciated. Thanks in advance for your time! Mary-Jacque Mann
One further comment after Michael's excellent summary.
If you can see any apoptotic cells in the sections, you shoud try and label these same cells using neighbouring sections. If these don't stain, you have a problem. You might also try and beg/borrow a Lowicryl embedded sample known to have reacted positively for an apoptotic marker, so that at least you can see if your antibody works. But really, apoptosis is such a rapid event that ultrastructural ID is perhaps as good as labelling, depending of course on why you want to label them. I seem to remember that apoptosis is complete in 20 mins. Shoot me down if I'm wrong!
Diana --
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Jay, Sorry, I forgot one thing! Tobias Baskin´s proposal for a MT fix sounds good as well. EGTA seems to me useful. But anyway, what kind of fixative you will choose, DON`T forget to keep your fixative at room temperature at least, this is a crucial prerequisite.
Michael ______________________________________________________________________________ Die Fachpresse ist sich einig: WEB.DE 18mal Testsieger! Kostenlos E-Mail, Fax, SMS, Verschlüsselung, POP3, WAP....testen Sie uns! http://freemail.web.de
Hello Jay! Try it with a fixative containing glut and tannic acid (TA), followed by osmification and embedding in epoxy. There is a classical paper:
Lewis G. Tilney et al.: "Microtubules: Evidence for 13 Protofilaments", JCB 59,pp 267-275, 1973
Tilney used 2% Glut and 2-8% tannic acid made in phosphate buffer. 1% digitonin was added to faciitate TA penetration.
My recipe is the following: make a sticky TA solution: solve 25 grs TA powder in 25 ml aqua, heat on a stirrer until you have a dark brown solution, be quick, so it will remain fluid. Filter with a paper filter . The solution made is stable over years, the older it gets the better your result. For fixation add 1-2% v/v of the stock TA solution to a cacodylate/phosphate buffer containing 4% Formaldehyd and 2% Glutaraldehyde. May be you will have to add 0,5-1% digitonin, but TA from the stock solution penetrates not that bad as directly in the fix soluted TA powder will do. Don´t fix your samples too long, because overcontrasting can occur (max. 24 hours). Then, follow a standard embedding protocol including osmificartion and uranylic acetate en-bloc staining. Try a quick dehydration with aceton to avoid tissue component extraction but I have satisfactorily results with normal alchoholic dehydration.
Hope this helps!
Good luck, with best regards, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I
Jay wrote } Hello All, } Can anyone suggest/recommend a fixation protocol that preserves } microtubules and/or centrioles without needlessly extracting other } cellular components?
______________________________________________________________________________ Die Fachpresse ist sich einig: WEB.DE 18mal Testsieger! Kostenlos E-Mail, Fax, SMS, Verschlüsselung, POP3, WAP....testen Sie uns! http://freemail.web.de
To the person looking for ultrastructural evidence of apoptosis: Take a look at one of my papers: American Journal of Pathology, Vol. 156, No. 6, June 2000. Good Luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
here we are, trying to find out how to face a SEM work related to a request from someone who manufactures some kind of synthetic boards to build up ceilings. We do not know much about this product but, they ensure it does not contain asbestos. Actually, they need to proof that!.
How would you make up the measurements in order to garantee or proof that no asbestos at all are found in these boards. It means a kind of "negative experiment" what we are asked to do.
Any experiences or sugestions?. We are looking at standards or regulations but not found about the sampling and measuring (observation) procedure for a negative result.
Thanks in advance,
Silvia Montoro Centro Regional de Investigación y Desarrollo de Santa Fe Güemes 3450 3000 Santa Fe Fax +54 - 342 - 455 0944 e-mail: csedax-at-arcride.edu.ar smontoro-at-arcride.edu.ar npratta-at-arcride.edu.ar
I was recently asked who manufactures cryo stage/prep equipment for SEM's. We have had such equipment for several years, but I have no idea what other suppliers are out there. Any help? Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
I want to say thanks to all people who replied to my inquiry on who can do EDS. I received 17 responses and/or suggestions. I was overwhelmed with the response. I have passed the information on to the investigator who will decide what direction she wants to go in.
I can't tell you how many times the listserver has come to the recuse. Sometimes a response comes while I'm thinking about asking the listserver. It like you are reading my mind. Besides being a lifesaver, it is also educational. I learn alot from all of you. Thanks again.
George
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 Fax 214-648-6408 eMail: George.Lawton-at-UTSouthwestern.edu
I posted a message to say that we have a Gatan Dual Ion Mill to dispose of. We have been swamped with replies. I had no idea what I was letting myself in for. Moreover, I got flu and so have been unable, til now, to respond. So, please no new contacts. Also those who have made contact already, please be patient. I will get to deciding what to do as soon as I can.
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Emitech makes several models. You can read about them in our on online: Home} Contents} K4 Disclaimer: ProSciTech supplies these instruments, but only within Australasia south of Singapore. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, February 15, 2001 5:32 AM, Bill Chissoe [SMTP:wchiss-at-ou.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I was recently asked who manufactures cryo stage/prep equipment for } SEM's. We have had such equipment for several years, but I have no idea } what other suppliers are out there. Any help? } Bill } } -- } ============================================================= } Bill Chissoe III } Electron Microscopist } University of Oklahoma } 770 Van Vleet Oval } Norman, Ok. 73019 } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } ============================================================= } }
Does anyone out there have experience in cleaning up fecal material so it can be processed for TEM. We are looking for a virus which we are going to label. Any help, anyone could give me ASAP would be greatly appreciated
Marie Jenkins Lab Manager Department of Environmental & Toxicologic Pathology Armed Forces Institute of Pathology Bldg 54 Rm M093B Washington, D.C. 20306-6000 Tel:202-782-2734 Fax: 202-782-9215 jenkins-at-afip.osd.mil
In a rationalisation of our institutes library we have come across two old volumes of the Illustrated Annual of Microscopy dated 1898 and 1900 published by Percy Lund, Humphries & Co London. Has anyone any information on the scarcity of these volumes so we can decide whether to donate them to a library with better archival storage facilities.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
Gatan (incorporating the cryo EM bit of Oxford Instruments), VG Microtech (Polaron), EmiTech and Bal-Tech are main players. Their contact details can be found in the Microscopy Vendors Database
http://www.kaker.com/mvd/list.html Chris
Date sent: Wed, 14 Feb 2001 11:31:41 -0800 } From: Bill Chissoe {wchiss-at-ou.edu}
We often get dust and bits of board from the site here to examine. We use powder X-ray diffraction to test for asbestos. If you have a theta-2theta diffractometer you could use, I can give you a protocol.
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi dear microscopists, } } here we are, trying to find out how to face a SEM } work related to a request from someone who manufactures some } kind of synthetic boards to build up ceilings. We do not know much } about this product but, they ensure it does not contain asbestos. } Actually, they need to proof that!. } } How would you make up the measurements in order to guarantee or } proof that no asbestos at all are found in these boards. It means a } kind of "negative experiment" what we are asked to do. } } Any experiences or suggestions?. We are looking at standards or } regulations but not found about the sampling and measuring } (observation) procedure for a negative result. } } Thanks in advance, } } Silvia Montoro } Centro Regional de Investigación y Desarrollo de Santa Fe } Güemes 3450 } 3000 Santa Fe } Fax +54 - 342 - 455 0944 } e-mail: csedax-at-arcride.edu.ar } smontoro-at-arcride.edu.ar } npratta-at-arcride.edu.ar
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
Dear Everybody, Our central science lab is trying to egt a coater unit, can any body suggest where to get one and what are the performance xteristics that goes with it also all neccesary accessories. thank You
Mr. O. O. ILORI DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING OBAFEMI AWOLOWO UNIVERSITY, ILE-IFE, OSUN STATE NIGERIA.
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Bill, Gatan has recently taken over the SEM cryostage previously marketed by Oxford Instruments (originally owned by Hexland in the 80's). My understanding is that the deal was the division in it's entirety which means that the personnel involved with tech support, sales, etc. also moved on to Gatan. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
On Wednesday, February 14, 2001 2:31 PM, Bill Chissoe {wchiss-at-ou.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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To analyze for asbestos at least two approaches can be used. A small piece of the sample can be pulverized and characterized by PLM. A more complete method that will provide some quantitative measure is to pulverize a measured mass of the material and then put into a DI solution and sonicate. Aliquot of the solution are then redeposited onto special filters via vacuum filtration and then prepped for analysis by TEM/EDS. There are many labs in the USA that specialize in these types of analysis. You can contact them to get more detailed information on these types of analysis.
Hope this helps.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
In America, there are lots of commercial asbestos testing labs. The analysis is usually performed by utilizing a TEM and/or a PLM based on protocols by NVLAP or other agencies. In your case, the most reliable +/- confirmation can be done with a TEM in 10 minutes, including sample preparation, should you have the permission to do asbestos analysis and be familiar with the morphology/structure(SAED)/chemistry(EDS). But I would like to suggest you send the sample out for the analysis because of the HAZMAT issue. It costs $60-$200 for one sample. For further information, you can contact me off line.
Disclaimer: I'm not doing asbestos analysis any more though had done it for a living for a couple of years, and nor do I have any financial interests in this kind of analysis.
-cy, Scientist, Rodel Inc.
-----Original Message----- } From: "csedax-at-alpha.arcride.edu.ar"-at-sparc5.microscopy.com [mailto:"csedax-at-alpha.arcride.edu.ar"-at-sparc5.microscopy.com] Sent: Wednesday, February 14, 2001 12:38 PM To: Microscopy-at-sparc5.microscopy.com Cc: csedax-at-ARCRIDE.EDU.AR
Hi dear microscopists,
here we are, trying to find out how to face a SEM work related to a request from someone who manufactures some kind of synthetic boards to build up ceilings. We do not know much about this product but, they ensure it does not contain asbestos. Actually, they need to proof that!.
How would you make up the measurements in order to garantee or proof that no asbestos at all are found in these boards. It means a kind of "negative experiment" what we are asked to do.
Any experiences or sugestions?. We are looking at standards or regulations but not found about the sampling and measuring (observation) procedure for a negative result.
Thanks in advance,
Silvia Montoro Centro Regional de Investigación y Desarrollo de Santa Fe Güemes 3450 3000 Santa Fe Fax +54 - 342 - 455 0944 e-mail: csedax-at-arcride.edu.ar smontoro-at-arcride.edu.ar npratta-at-arcride.edu.ar
Thanks to all who answered my inquiry about cryoSEM. As always, this is a very friendly, helpful group. Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
It is my understanding that all asbestos testing with an Electron Microscopy Lab must acquire certification from not only their state health agency but also there are Environmental Protection Agency regulations, one must thoroughly examine. I would be very cautious to accept any work involving Asbestos testing. Several laboratories have been sued. We were also approached by a local company in Rochester, NY who would provide us with a substantial amount of money to test asbestos, but upon talking to the NY state Health Dept., and looking into the Technician's certification course(which is not cheap and is 1year), and the laboratory would also have to be licensed, we decided it wasn' t going to be worth it.
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core
STOOL Samples of about 2-4 gm can be collected into a sterile container filled only half way. Make a 10-20% suspension in distilled water and clarify by centrifuging at 3000 rpm for 30 minutes. Then take the supernatant and centrifuge again at 15,000 RPM for one hour to concentrate a viral laden pellet. This pellet you could resuspend in various dilutions and drop onto a coated grid and immunolabel.
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core
-----Original Message----- } From: "JENKINS-at-afip.osd.mil"-at-sparc5.microscopy.com [mailto:"JENKINS-at-afip.osd.mil"-at-sparc5.microscopy.com] Sent: Wednesday, February 14, 2001 8:34 PM To: microscopy-at-sparc5.microscopy.com
Does anyone out there have experience in cleaning up fecal material so it can be processed for TEM. We are looking for a virus which we are going to label. Any help, anyone could give me ASAP would be greatly appreciated
Marie Jenkins Lab Manager Department of Environmental & Toxicologic Pathology Armed Forces Institute of Pathology Bldg 54 Rm M093B Washington, D.C. 20306-6000 Tel:202-782-2734 Fax: 202-782-9215 jenkins-at-afip.osd.mil
The official deadline for submission of papers for the 2001 Microscopy and Microanalysis meeting is today, February 15. However, there have been some problems with the database so the deadline is being extended until Noon Eastern Time on Monday, February 19. All Title, Author and AV requirement data must be entered into the database at mmconference.org/2001 by that time and papers must be received at the following address by Tuesday, February 20.
MSA Meeting Managers 7000 West Southwest Highway Chicago Ridge, IL 60415
877-672-6271 (Telephone)
If you have problems submitting your Author information, or if you would like to submit a paper for the late breaking poster session after February 19, please contact Bob Price (Price-at-med.sc.edu) or 803-733-3393
Thanks.
Bob Bob Price M&M 2001 Program Chair 803-733-3393 (T) 803-733-1533 (F)
Manager Keck Center for Characterization of Nanoscale Soft Materials-Northwestern University seeks a Ph.D. scientist with experience in operation of imaging TOF-SIMS, high vacuum STM/AFM and high throughput XPS instruments to manage open user facility. For detailed job description see http://www.chem.nwu.edu/group_openings.htm. Applicants should send CV and summary of relevant experience and qualifications to Dr. Robert N. Scott, Department of Chemistry, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208-3113 or to r-scott-at-chem.nwu.edu. EEO/AA Employer.
There is a paper in Applied Surface Science 167 (2000), pp. 59-68 entitled, "Structural studies of amorphous and crystallized tungsten nitride thin films by EFED, XRD, and TEM" by Y.G. Shen and Y.W. Mai that you should look up if you are interested in this topic.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Yan Xin [mailto:xin-at-magnet.fsu.edu] Sent: Monday, February 12, 2001 12:32 PM To: microscopy-at-sparc5.microscopy.com Subject: software for RDF of amorphous materials
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I am very interested in getting softwares to process electron diffraction patterns from disordered materials to obtain the G(r) (reduced intensity function), and J(r) (radial distribution function).
Any information on how to get these softwares is appreciated. In particular, information on free ones are most welcome.
Regards Yan Xin
======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
We are in the process to get a confocal microscope for a multi-user center. The major application for this microscope will be using green fluorescent proteins and /or styrul dyes (e.g., FM 1-43) to label specific neuronal pathways, including living cells.
Our consideration will be either "Leica TCS SP2" or "Zeiss LSM 510", any suggestion and advice on the comparison of these two microscopes will be highly appreciated.
Many thanks.
******************************************* Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4. 214A Dallas, TX 75390-9111
WE have the Zeiss LSM 510 and it's great. Their customer support/technical support is outstanding.
Good Luck- Michelle Taurino Aventis Pharmaceuticals Senior Scientist Bioimaging and Molecular Histology Tel-908-231-3357 Fax-908-231-3962 e-mail: Michelle.Taurino-at-aventis.com
-----Original Message----- } From: Xinran Liu [mailto:xinran.liu-at-UTSouthwestern.edu] Sent: Thursday, February 15, 2001 7:50 PM To: Microscopy-at-sparc5.microscopy.com
We are in the process to get a confocal microscope for a multi-user center. The major application for this microscope will be using green fluorescent proteins and /or styrul dyes (e.g., FM 1-43) to label specific neuronal pathways, including living cells.
Our consideration will be either "Leica TCS SP2" or "Zeiss LSM 510", any suggestion and advice on the comparison of these two microscopes will be highly appreciated.
Many thanks.
******************************************* Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4. 214A Dallas, TX 75390-9111
At 6:50 PM -0600 2/15/01, Xinran Liu wrote: } } } We are in the process to get a confocal microscope for a multi-user center. } The major application for this microscope will be using green fluorescent } proteins and /or styrul dyes (e.g., FM 1-43) to label specific neuronal } pathways, including living cells. } } Our consideration will be either "Leica TCS SP2" or "Zeiss LSM 510", any } suggestion and advice on the comparison of these two microscopes will be } highly appreciated. } } Many thanks. } **************************************
I cannot say anything about the Leica instrument, since I have never used it. I can however, speak about the Zeiss LSM 510. I operate a multi-user core facility and we have had an LSM 510 here since Nov. 1998. It has proven itself to be a reliable and fairly robust instrument. We have aver 50 registered users. Some work quite frequently, others rarely. Our regular users have all been trained to use the system and work independently. The types of samples examined also vary widely, from immunolabelled tissue sections (paraffin and cryo) to live cell culutres expressing GFP. Overall, we are very happy with it.
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I have had several requests for the asbestos protocol, so here it is.
1) Immerse sample in 10% HCL for 30 mins or until bubbles have stopped. (This removes filler material, usually calcite ICPMS 5-586.)
2) Decant and wash in water.
3) Evaporate till dry.
4) Grind to a powder using pestle and mortar. (Obviously, if the sample is suspected to be asbestos, take suitable precautions!)
5) Mount as a standard thick powder sample for theta-2theta diffractometry
6) Obtain data from 5-60 degrees (theta) using Cu K alpha radiation. (We have a fairly standard Philips diffractometer, with graphite monochromator and acceptor slit.)
In the UK, the notifiable asbestos compounds according to act of Parliament (don't ask me which one!) are:
Other compounds which may be considered to be hazardous/asbestos-like are: Quartz (SiO2) ICPMS 33-1161 Tridymite (SiO2) ICPMS 18-1170 or 14-260 Christobalite (SiO2) ICPMS 11-695 Kaolinite ICPMS 14-164 Amosite ICPMS 29-1170 Anorthite ICPMS 18-1202 Phlogopite ICPMS 10-492
You will need the ICPMS database and the ability to compare and/or search for matches.
Detection limit is about 5%, depending on the other materials in the sample.
This is not an accredited test. If you have a sample which doesn't show up positive, it may still contain asbestos - if in doubt, get an accredited test house to check.
No liability is accepted or implied in providing this information.
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
I have been looking for a copy of "Food Microscopy", by Olga Flint. It is one volume of the Royal Microscopy Society handbook series. I saw copies available during the last MSA meeting, but cannot recall who the vendor was. Can somebody point me towards a company or individual who may have a copy of this book available for sale?
Karl Hagglund, Researcher P&G Food and Beverage Analytical and Microbiology 6071 Center Hill Ave., Box 117 Cincinnati, OH 45224 (513)634-0146
I have been looking for a copy of "Food Microscopy", by Olga Flint. It is one volume of the Royal Microscopy Society handbook series. I saw copies available during the last MSA meeting, but cannot recall who the vendor was. Can somebody point me towards a company or individual who may have a copy of this book available for sale?
Karl Hagglund, Researcher P&G Food and Beverage Analytical and Microbiology 6071 Center Hill Ave., Box 117 Cincinnati, OH 45224 (513)634-0146
The US Environmental Protection Agency (US EPA) developed a series of methods [Polarized Light Microscopy, Transmission Electron Microscopy, Gravimetry (including acid dissolution and ashing), and X-ray Powder Diffraction] for the analysis of asbestos in building materials that are described in the following document:
EPA 600/R-93-116 Method for the Determination of Asbestos in Bulk Building Materials. July 1993. (NTIS / PB93-218576) [Updates and replaces Interim version in 40 CFR 763 Subpart F App A]
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 100 Bureau Drive, Stop 8371 Gaithersburg, MD 20899-8371 http://www.nist.gov/cstl/div837/837.02/
We also have a multi-user facility and have two Leica systems, one an inverted with multi-photon lasers and the other an upright with argon, krypton, and HeNe lasers. When they work, they're great. However, software has been less than desirable , as has service. Software keeps being "upgraded" but not necessarily improved. I'm sure all the instruments have problems but I think we've had more than our share with Leica.
At 06:50 PM 2/15/01 -0600, Xinran Liu wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
I have (not) identified asbestos... That is, some insulating material was brought to me with the question, "Is it asbestos?". I was able to say that it was not since careful EDS examination of the powdered material revealed no trace of the elements associated with asbestos. It was 100% gypsum.
Even if not certified, this would seem to produce a satisfactory answer. ..Or are the regulations now so absurd that officialdom is concerned someone may mistake brass for asbestos? ;)
Woody White McDermott Technology, Inc. http://www.mtiresearch.com/
Need assistance on Gatan PIPS. Believe DP and MDP are functioning properly, but specimen chamber vacuum is "glitchy."
Symptoms: DP light on constant, MDP light never comes on, DP test reads 19.9 at all times, Lights on vac and vent do not glow nor will sample raise or lower.
Please respond off list.
J. Cindy Kleier Specimen Preparation Engineer Northwestern University Evanston, IL. USA 847-491-7807 j-kleier-at-northwestern.edu
fatbrain.com and barnes and noble and amazon.com all list this book for 20 something bucks. hope this helps...
ed sharpe archivist for SMECC
{ { Subj: help me to find a book Date: 2/16/01 2:06:41 PM US Mountain Standard Time From: hagglund.kw-at-pg.com-at-sparc5.microscopy.com To: microscopy-at-microscopy.com
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I have been looking for a copy of "Food Microscopy", by Olga Flint. It is one volume of the Royal Microscopy Society handbook series. I saw copies available during the last MSA meeting, but cannot recall who the vendor was. Can somebody point me towards a company or individual who may have a copy of this book available for sale?
Karl Hagglund, Researcher P&G Food and Beverage Analytical and Microbiology 6071 Center Hill Ave., Box 117 Cincinnati, OH 45224 (513)634-0146
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We are e-mailing to let you know that our textbook:
"Transmission Electron Microscopy and Diffractometry of Materials"
has just been published by Springer. This textbook and its earlier drafts have been used for over a decade in courses on transmission electron microscopy (TEM) and x-ray diffractometry (XRD) at the California Institute of Technology and University of Virginia. These courses are taken by graduate students and advanced undergraduates in materials science, solid-state physics, and solid-state chemistry.
The book emphasizes themes common to both diffraction and microscopy, such as wave coherence, scattering from atoms, and the formation and analysis of diffraction patterns. It also describes the uniqueness of TEM and XRD. The book has 748 pages through the index, includes approximately 500 accompanying illustrations, and a total of 151 problems at the ends of chapters. The Appendix provides a set of introductory TEM laboratory exercises, and contains up-to-date reference data for both TEM and XRD. The chapters are:
1. Diffraction and the X-Ray Powder Diffractometer, p. 1 2. The TEM and its Optics, p. 63 3. Scattering, p. 123 4. Inelastic Electron Scattering and Spectroscopy, p. 167 5. Diffraction from Crystals, p. 225 6. Electron Diffraction and Crystallography, p. 275 7. Diffraction Contrast in TEM Images, p. 339 8. Diffraction Lineshapes, p. 423 9. Patterson Functions and Diffuse Scattering, p. 467 10. High-Resolution TEM Imaging, p. 523 11. Dynamical Theory, p. 595 12. Bibliography, p. 661 13. Appendix, p. 675 14. Index, p. 735
To help with teaching, sections containing specialized or higher-level material are marked, and paths around these sections are suggested. A secure web site has been established with worked solutions to 3/4 of the problems in the text.
A web site with excerpts from many chapters is:
http://www.caltech.edu/~matsci/btf/TEM_Book.html
Publication and order information can be found at the Springer site:
We hope that you find our book useful and look forward to hearing from you about it.
Sincerely yours, Brent Fultz and Jim Howe
Brent Fultz, Professor of Materials Science California Institute of Technology, mail 138-78 Pasadena, CA 91125, USA Office: 626-395-2170; Fax: 626-795-6132; E-mail: btf-at-hyperfine.caltech.edu
James M. Howe, Professor and Director of the Electron Microscope Facility Department of Materials Science & Engineering University of Virginia Charlottesville, VA 22904-4745, USA Office: 804-982-5646; Fax: 804-982-5660; E-mail: jh9s-at-virginia.edu
Hello, We are looking for some Wild 20x oculars to equip a Wild M5 stereo microscope. If anyone has any extras or knows of a source, please respond directly to: LMM7001-at-humboldt.edu
Thanks, Lewis mcCrigler Equipment Tech. Humboldt State University
I think this is fundamentally an EDS detection limit issue. Question is when you see Ca, S, and possibly O peaks only on a spectrum from your insulation, how sure you can be to say that you don't have 1 vol% of Si-Mg-Fe-Mn etc while the 1 vol% is the cut-off concentration in the US regulation governing asbestos analysis of building materials. Personally, I wouldn't be very comfortable with an EDS un-identification of asbestos.
-cy, Ex-expert of asbestos analysis
-----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-mcdermott.com] Sent: Friday, February 16, 2001 4:02 PM To: 'Microscopy-at-MSA.Microscopy.Com'
Any comments on this one?
I have (not) identified asbestos... That is, some insulating material was brought to me with the question, "Is it asbestos?". I was able to say that it was not since careful EDS examination of the powdered material revealed no trace of the elements associated with asbestos. It was 100% gypsum.
Even if not certified, this would seem to produce a satisfactory answer. .Or are the regulations now so absurd that officialdom is concerned someone may mistake brass for asbestos? ;)
Woody White McDermott Technology, Inc. http://www.mtiresearch.com/
Forgot to mention that my previous comments only apply to EDS/SEM that appears to be what Mr. Woody meant. In the case of TEM analysis, Woody should be 100% right if the beam is focused on each individual fibers and sufficient number of fibers are checked.
-cy, Ex-fiber counter on TEM, PLM and PCM
-----Original Message----- } From: Ni, Chao-Ying Sent: Saturday, February 17, 2001 5:48 PM To: 'White, Woody N.'; 'Microscopy-at-MSA.Microscopy.Com'
Any comments on this one?
I have (not) identified asbestos... That is, some insulating material was brought to me with the question, "Is it asbestos?". I was able to say that it was not since careful EDS examination of the powdered material revealed no trace of the elements associated with asbestos. It was 100% gypsum.
Even if not certified, this would seem to produce a satisfactory answer. .Or are the regulations now so absurd that officialdom is concerned someone may mistake brass for asbestos? ;)
Woody White McDermott Technology, Inc. http://www.mtiresearch.com/
We recently had a Leica TCS SP2 installed, and it has been running just fine, software is easy to use and service has been great. In a previous institution we had two Leica TCS's - hardware fine, software OK, service also great.
IMHO, the main considerations are first, service and reliability - and service varies quite a bit with location for each company. Second, do you really need the SP function of the Leica? If not, I think these two instruments are much of a muchness for routine work. There are pluses and minuses on each side and you need to decide what your priorities are. Third, what kind of price/package can you negotiate?
Another 2¢ worth..... cheers,
At 06:50 PM 2/15/01 -0600, Xinran Liu wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Hello all, I need to do some immunogold TEM with antibodies that work well for light microscopy immunohistochemistry but require various antigen retrieval treatments. I used to do a lot of immunogold in the old pre-antigen retrieval times. I wonder what are the current developments in this area. Are antigen retrieval techniques used on LR White sections? Thanks for any suggestions,
Sarka Lhotak, PhD.
Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
I understand the situation. In addition to "bulk" analysis, numerous locations were examined using higher and higher BSE magnifications to sort out any (non existent in this case) Z differences. EDS was used for each as well. In addition, with the appropriate choice of accelerating potential and count integral, my EDS could easily detect { 0.25%. Granted, this is not a very efficient method and if ANY of the questionable elements were detected, nothing is proved.
Woody
} Dear Woody, } } I think this is fundamentally an EDS detection limit issue. } Question is when } you see Ca, S, and possibly O peaks only on a spectrum from } your insulation, } how sure you can be to say that you don't have 1 vol% of } Si-Mg-Fe-Mn etc } while the 1 vol% is the cut-off concentration in the US } regulation governing } asbestos analysis of building materials. Personally, I } wouldn't be very } comfortable with an EDS un-identification of asbestos. } } -cy, Ex-expert of asbestos analysis } } } } -----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-mcdermott.com] } Sent: Friday, February 16, 2001 4:02 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: RE: Asbestos protocol } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } Any comments on this one? } } I have (not) identified asbestos... That is, some insulating } material was } brought to me with the question, "Is it asbestos?". I was } able to say that } it was not since careful EDS examination of the powdered } material revealed } no trace of the elements associated with asbestos. It was } 100% gypsum. } } Even if not certified, this would seem to produce a } satisfactory answer. } ..Or are the regulations now so absurd that officialdom is concerned } someone may mistake brass for asbestos? } ;) } } Woody White } McDermott Technology, Inc. } http://www.mtiresearch.com/ } } http://home.att.net/~woody.white }
I would like to thank everyone who sent suggestions for cleaning the dust from a Polaroid DMC digital camera. We had great luck using a lint-free cloth moistened with methanol and very gently wiping the filter surface after opening the shutter. Practically all of the "spots" we were seeing on our digital images were successfully removed and the camera is now capable of meeting our most demanding needs. Polariod really came through with excellent technical support via the Listserver. If anyone is interested in learning more details (I know some of you mentioned you had the same "problem"), please feel free to contact me and thanks again for all your help.
Michael Simko Research Manager Metallography Group U. S. Steel Research and Technology Center
I wanted to thank everyone who sent suggestions for cleaning the dust from a Polaroid DMC digital camera. We had great luck using a lint-free cloth moistened with methanol and very gently wiping the filter surface after opening the shutter. Practically all of the "spots" we were seeing on our digital images were successfully removed and the camera is now capable of meeting our most demanding needs. Polariod really came through with excellent technical support via the Listerserver. If anyone is interested in learning more details (I know some of you mentioned you had the same "problem"), please feel free to contact me and thanks again for all your help.
Michael Simko Research Manager Metallography Group U. S. Steel Research and Technology Center
I am trying to find some information on staining mitochondria with Janus green, including the staining mechanism. However nothing relevant seems to be available in the internet or in my technical books. This is mainly for teaching purposes.
Can anyone send me information on the subject and/or relevant URL(s) to consult ?
Thanks for your help Dr. A.P. Alves de Matos apmatos-at-ip.pt
Food Structure & Functionality Symposium 2001 An international symposium leading food structure & Functionality studies through the 21st century
"webaddress at the AOCS site http://www.aocs.org/member/division/fsff/index.htm"
Being held at the 92nd AOCS Annual Meeting & Expo, May 13_16, 2001, Minneapolis Convention Center, Minneapolis, Minnesota, USA. For information, e_mail us at meetings-at-aocs.org
The symposium has two themes: * New and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure_function relationships in foods; * Food system studies covering any part of the processing chain _ from the raw material to the final product, including trouble shooting. --------------------------------------------------- Tentative Program (confirmed as of February 19th, 2001)
May 13th _ Short Courses (short courses will run for a full day, and will run concurrently)
Short Course #1 Food Contaminants. Course organiser: Mark Auty (mauty-at-moorepark.teagasc.ie)
Short Course #2 Understanding structure-function relationships in food systems through specific localisation methods and microscopy.
For more information contact Marcel Paques (paques-at-nizo.nl) ---------------------------------------------------
May 14th-16th inclusive -Technical sessions (6 sessions will run over 3 days)
Monday, May 14th, 2001 Morning Symposium opening Plenary lecture _ Food Quality and why the Structure matters P. Lillford, Unilever Research, Colworth House UK
Session 1: Dairy Products and Fat Based Foods Session _ chairs M. Auty and M. Paques
Milk protein polysaccharide interactions in aqueous solutions and oil-in-water emulsions. H. Singh*, Y. Hemar & P. Munro, Institute of Food, Nutrition and Human Health, Massey University, Palmerston North, New Zealand
Structural functions of dairy ingredients in products formulated with taro flour. C. Onwulata USDA/ARS,Eastern Regional Research Center,Wyndmoor, PA 19038
Confocal imaging of galactomannan mode of action in recrystallisation of ice in model ice cream D. Ferdinando, Unilever Research Colworth House, UK
Heating of Food Proteins in a Closed System at High Temperature N. Kitabatake, Kyoto University, Japan
Milk Protein -molecular components and functional properties N.C. Ganguli ,Indian Dairy Association
Afternoon Session 2: Food Safety _ chairs J. W. Arnold and R. Droleskey Prevention of Foodborne Illness Through Sanitation and Processing Technology J.W. Arnold; USDA/ARS, Russell Research Center; Athens, GA 30604
PreHarvest Intervention Strategies to Control Foodborne Pathogens in Poultry J. A. Byrd; USDA/ARS/SPARC; College Station, TX 77845
Interactions of Competitive Exclusion Cultures with the Intestinal Mucosa of Newly Hatched Chicks R. Droleskey; USDA/ARS/SPARC; College Station, TX 77845
Adhesion of Salmonella on Alfalfa Sprouts A. Chartowski; USDA/ARS, Western Regional Research Center; Albany, CA 94710
Growth of Fusarium moniliforme Dependent upon Corn Tissue Type I. E. Yates; USDA/ARS, Russell Research Center; Athens, GA 30604
Dedicated poster viewing 4:00_6:00PM
Evening: Round Table Discussion _ topic to be announced
--------------------------------------------------- Tuesday, May 15th, 2001 Morning Session 3: New Methods and Techniques for Food Structure and Functionality Analysis _chair K.Groves Applications of Scanning Near Field Optical Microscopy in the analysis of food and food-related materials. A.R. Kirby*, P. Gunning, P.J. Wilde and V. J. Morris, Institute of Food Research, Norwich, England
Diffusing wave spectroscopy - a new and non-invasive method for the investigation of the structure, dynamics and interactions in complex food systems M. Alexander* and P. Schurtenberger, Universite de Fribourg, Switzerland
AFM as a tool for characterising polysaccharides and their modification. V.J. Morris*, A.P. Gunning, A.R. Round, E. Adams, E. Kroon and G. Williamson, Institute of Food Research, Norwich, England
Micro-rheology - functional properties of food systems and their origin in microstructure M. Paques, Y. Nicolas, G. van Aken, H. Tromp, A. Janssen, Unilever Research Vlaardingen, The Netherlands
Immunolocalization of Transgenic Protein in Wheat Endosperm M. L. Parker (invited),* E. Stoger, *R. Casey and *P. Christou, Institute of Food Research, Norwich, England and *John Innes Centre, Norwich, England
Food: How complex can it be? E. Esselink ,Unilever Research Vlaardingen, The Netherlands
Multiple Extrusion Module for determining changes in consistency during working - a new method. J.F.C. Van Maanen*, B. Dunnewind, and A. Jurgens. TNO Nutrition and Food Research Institute, The Netherlands
Specific detection of components and compounds through microscopical immunodetection J. Leunissen, AURION: Immuno-Gold Reagents & Accessories, The Netherlands
Food Structure and Functionality Division Luncheon. Dr. Brian Brooker (Institute of Food Research, England, retired), the recipient of the Food Structure and Functionality Division Award, will give a presentation entitled: Fat crystals - the importance of being small.
Afternoon Session 4: Agricultural Applications of Microscopy and Imaging Session chairs D.F.Wood and P. Allan-Wojtas
The Utility of Sorting in Agriculture. H. J. Arnott, Department of Biology, University of Texas at Arlington, Texas
The Potential for Automatic X-ray Sorting of Insect Infested Grain R. Haff . USDA - ARS -WRRC, Albany, California.
Automated Sorting of Almonds with Embedded Shell by Laser Transmittance Imaging T. Pearson* R. Young, USDA -ARS- WRRC, Albany, California.
Use of a GFP-transformed strain of Fusarium graminearum to study ear rot development in corn S. S. Miller. , AAFC-ECORC, Ottawa, Ontario, Canada.
Popping modifies endosperm structure and improves digestibility in maize and sorghum. M.L. Parker, Institute of Food Research, Norwich, England.
Microstructural Changes in Rice During Cooking D. Wood* and P.C. Yu . USDA -ARS- WRRC, Albany, California.
The Food Structure and Functionality Division Member meeting will be held immediately following this session. All are welcome.
--------------------------------------------------- Wednesday, May 16th, 2001 Morning Session 5: Ingredients and Food Processing chairs D. Kittleson and J. Charbonneau
Water continuous fat crystal networks in ice cream induced by unsaturated monoglycerides N. M. Barfod , Danisco Cultor, Brabrand, Denmark
High pressure application of food systems and its impact on functional ingredients B. Tauscher*, P. Butz,and A.F. Garcia, Federal Research Center for Nutrition, Karlsruhe, Germany
Specificity and application of lipolytic enzymes in bread making processes T.B. Frandsen, T. Spendler, G. Budolfsen, L. Christiansen and J. B. Neilsen, Novozymes A/S, Bagsvaerd, Denmark
The development of bubble structure in bread doughs. M.B. Whitworth and J.M. Alava, Campden & Chorleywood Food Research Association, UK
Structure-texture relationships in heat resistant chocolate K. Groves, P. Subramaniam and O. Murphy, Leatherhead Food Research Association, UK
Reduction of oil uptake by methyl-cellulose coatings applied to fried dough M. A. Garcia, C. Ferrero, N. Bertola, M. Martino* and N. Zaritzky, CIDCA CONICET. Fac. de Ciencias Exactas and Fac. de Ingenieria, Universidad Nacional de La Plata, Argentina.
Afternoon Session 6: Colloidal and Interfacial Sciences _ chairs D. Pechak and M. Paques
Rheology and Structure of Particulate Protein Gels T. van Vliet, Wageningen Centre for Food Sciences,Wageningen, The Netherlands
Time-temperature studies of food polymer gelation. Paulson, AT., Nickerson, M.T. and Speers, R.A. Department of Food Science and TechnologyDalhousie University, Halifax, NS, Canada
--------------------------------------------------- Contact information for the chairs is shown below, in alphabetical order:
Paula Allan-Wojtas Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre, Kentville, Nova Scotia, Canada B4N 1J5 Tel: (902) 679-5566 FAX: (902) 679-2311 email: allanwojtasp-at-em.agr.ca
Judy Arnold USDA -ARS -RRC 950 College Station Rd. P.O. Box 5677 Athens, GA 30604-5677 USA Tel: (706) 546-3515 FAX: (706) 546-3068 email: jarnold-at-ars.usda.gov
Mark Auty Dairy Products Research Centre TEAGASC Moorepark, Femoy, Co. Cork Ireland Tel: 011-353-25-42447 FAX: 011-353-25-42340 email: mauty-at-moorepark.teagasc.ie
James E. Charbonneau National Food Processors Association Food Chemistry and Packaging Department 1401 New York Ave, NW Washington, D.C. 20005 USA Tel: (202) 639-5972 FAX: (202) 639-5991 email: jcharbo-at-nfpa-food.org
Kathy Groves Leatherhead Food Research Association Randalls Road, Leatherhead Surrey KT22 7RY England Tel: 44 0132 822330 FAX: 44 0132 386228 email: kgroves-at-lfra.co.uk
Tony McKenna New Zealand Dairy Institute Private Bag 11 029 Palmerston North, New Zealand Tel: 011 64 6 350 4649 FAX: 011 64 6 356 1476 email: tony.mckenna-at-nzdri.org.nz
Marcel Paques Wageningen Centre for Food Sciences/Unilever Research Vlaardingen P.O. Box 20, 6710 BA Ede The Netherlands Tel: 011 31 318 659690 FAX: 011 31 318 650400 email: paques-at-nizo.nl
David Pechak Kraft Technology Centre 801 Waukegan Road Glenview, IL 60025 USA Tel: (847) 646-4808 FAX: (847) 646-3864 email: dpechak-at-kraft.com
Delilah Wood USDA -ARS -WWRC 800 Buchanan Street Albany, CA 94710 USA Tel: (510) 559-5653 FAX: (510) 559-5777 email: wood-at-pw.usda.gov
Food Structure & Functionality Symposium 2001 An international symposium leading food structure & Functionality studies through the 21st century
"webaddress at the AOCS site http://www.aocs.org/member/division/fsff/index.htm"
Being held at the 92nd AOCS Annual Meeting & Expo, May 13_16, 2001, Minneapolis Convention Center, Minneapolis, Minnesota, USA. For information, e_mail us at meetings-at-aocs.org
May 13th - Short Courses (short courses will run for a full day, and will run concurrently)
Short Course #1 Food Contaminants. Most food manufacturing companies possess light microscopes, but seldom use them to their full potential. Microscopy is a powerful problem-solving tool for both contaminant analysis and food ingredient characterisation. This short course is aimed at Quality Control/Assurance and Food Product R&D personnel. The course comprises lectures and hands-on tuition from expert microscopists. The aim of the course is to provide delegates with a practical introduction to food microscopy, in particular contaminant (foreign body) identification and food ingredient characterisation.
Programme topics include: Getting the most from your light microscope - basic setting up and contrast techniques Preparing food samples for microscopy How to approach foreign body identification Identifying plant and animal remains Identifying glass, plastics, minerals Instrumental methods for contaminant analysis Microscopy in product research and development
Course contributors: John Shane (McCrone Research Institute, USA) Jim Charbonneau (National Food Processors Association, USA) Kathy Groves (Leatherhead Food Research Association, UK). Diana Kittleson (Pillsbury Technical Labs, USA) Mark Auty (Teagasc, Moorepark, Ireland) Course organiser: Mark Auty (mauty-at-moorepark.teagasc.ie)
Short Course #2 Understanding structure-function relationships in food systems through specific localisation methods and microscopy.
Spreadability, shelf life, fracture behaviour, creaminess, and mouthfeel are examples of functional properties of food products. These properties originate from the microscopic structure of the products. Specific localisation techniques and microscopy are powerful new tools to facilitate intelligent modification of ingredient composition or processing to obtain targeted product properties. The short course is aimed at R&D personnel in the Foods area (fundamental research, innovation, and product development). The course provides lectures and an intensive hands-on practical part enabling participants to gain sufficient basic knowledge and skills to set-up and implement the methods in their own work.
Programme topics include: D Introduction on specific localisation methods and principles D Localisation strategies, marking options, and imaging approaches D Experimental set-up, preparation and incubation procedures D Demonstration cases D Hands-on practical sessions
Course contributors: Jan Leunissen (Aurion: immuno Gold Reagents & accessories, custom labeling, The Netherlands) Gary Fulcher (University of Minnesota, USA) Paula Allan-Wojtas (Agriculture and Agri-Food, Canada). Marcel Paques (Wageningen Centre for Food Sciences/Unilever Research Vlaardingen, The Netherlands)
For more information contact Marcel Paques (paques-at-nizo.nl) ---------------------------------------------------
Technical sessions (6 sessions will run over 3 days) will follow the short courses from May 14th-16th inclusive.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
I'm not sure about this, but I recall someone telling me that the current Leica eyepieces for the MZ series of stereomicroscopes will also work on the older Wild M5, M8, etc.
It would be worth a shot to call your local Leica rep, or contact Leica Customer Service at 1-800-248-0123.
My name is Hunter O'Reilly. I recently received my Ph.D. in genetics from the University of Wisconsin-Madison in December 2000. I am also a contemporary artist currently working full-time on an art project called Radioactive Biohazard. Radioactive Biohazard is a project that integrates art and science to bring awareness of issues in biotechnology to the public in a positive light. A free catalog containing scientifically accurate yet simple explanations of the artworks will be available to the public.
I am very interested in using electron micrograph images of viruses (i.e. HIV, hepatitis, rabies, polio, herpes, ebola, adenovirus) in digital artworks for the second part of this project described below. I am also interested in other visually interesting microscopic images. Any artwork that used any part of a scientist's image would give the scientist credit for that part of the image both in the exhibit and publication of the artwork. Donations of images can also be made anonymously. I have high speed internet access. Very large files can be sent to me via email.
My artwork has appeared on the covers of several international scientific journals including Nature Reviews Genetics, Medical Genetics, The EMBO Journal, Promega's Neural Notes, and Developmental Dynamics. You can view some of these covers at http://www.artbyhunter.com/media/
Please email questions and/or microscopic images to Hunter O'Reilly at
hunter-at-artbyhunter.com
You can view my artwork on my website at http://www.ArtByHunter.com
Thank you for your time and consideration! Please read further for more information.
I have received donations of images from Biotechniques and the following individuals for use in this project.
Biotechniques: A Journal of Laboratory Technology for Bioresearch and Eaton Publishing Co. Dr. Peter Angeletti, Post-Doc at the McArdle Laboratory for Cancer Research Dr. James Briscoe, Columbia University Dr. James Ellingboe, Scientific Editor of Biotechniques Dr. Prakash Hande, Columbia University Dr. Mary McCarthy, Associate Scientific Editor of Biotechniques Dr. James A. Priess, Fred Hutchinson Cancer Research Center Dr. Alison Roberts, University of Rhode Island Dr. Charlotte Schubert, formerly at the Fred Hutchinson Cancer Research Center
Radioactive Biohazard will premiere at the Walker's Point Center for the Arts in Milwaukee, Wisconsin April 20, 2001.
The exhibit will consist of three parts: 1. oil paintings reinterpreting science as art exploring themes such as cloning and genetic identity--For example I am creating one oil painting on the theme of stem-cell research, and the potential to create organs for transplants from stem-cells. I am creating another oil painting regarding the use of human cloning as a treatment for infertility. The explanations in the catalog will be simple for the general public, but scientifically accurate. Controversial issues surrounding this research will also be discussed. I will write the original explanations, and have other scientists proofread and make suggestions before they are finalized. 2. enlargements of micrographs of cells, viruses and bacteria highlighted with neon tubing--These artworks will be digital collages of microscopic images. With some of the collages, I will create a landscape. With others I may superimpose the image of a person. In the catalog and next to each artwork, there will be a diagram siting the source of each part of the image, explaining what the image is, and possibly how the image is obtained. The creation of the digital collage will be artistic, but there will be scientific explanations to accompany the works. These artworks will allow the general public to see images they have never seen before. 3. a laboratory bench installation: an artistic interpretation of a laboratory bench, equipment etc.
You will find a longer description at http://www.artbyhunter.com/artgallery/radbio.html
My homepage is at http://www.artbyhunter.com
Thank you for your time and consideration! I look forward to hearing from you!
It's a couple of years since I did any antigen retrieval and then I did it on tissue pieces, which were labelled preembedding. I found citrate buffer heated to about 95deg was effective and the most gentle. Have no info on LR White sections, except that in my hands they never seem to label with anything!
Diana
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--
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
} From: {"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com} } Look in the phone book. } } Why must we endure these mundane questions, can anybody in this world think } for themselves.
Not all of us live in a city, state or even a country that has a microscope repair and service company.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
In some areas it may not be as simple as looking in the phone book. This listserver was meant to answer question not as a "put down' media.
Thank You,
Earl ----- Original Message ----- } From: {"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, February 19, 2001 3:52 PM
The question may have been unnecessary, but wasn't this response rather abrupt, unhelpful and unfriendly, particularly as it was anonymous?
} Look in the phone book. } } Why must we endure these mundane questions, can anybody in this world } think for themselves.
Have a good day.
Robin H Cross (Mr) Director : Electron Microscopy Unit Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/index.htm
** remember ICEM-15 in Durban in September 2002 **
} } Look in the phone book. } } Why must we endure these mundane questions, can anybody in this } world think for themselves. }
Very rude.
And coming from someone sheltering behind anonymity
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
A couple of hints. Here, a local microscope maintenance company (not listed in the phone book except in yellow pages under scientific instruments) comes around once a year and is known by several other company reps. Your microscope sellers may know of someone, as may other microscopy-related suppliers, even if you can't find anything listed in the phone book or on the web. The suppliers are usually unwilling to do this type of maintenance themselves, but there are local exceptions, so it's worth asking.
good luck,
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
I would like to know if anyone can recommend any books regarding the failure modes and mechanisms of plastics and ceramics. My searches have been yielded little (read as nothing), and my library is woefully lacking.
Thanks to all in advance for your help!
Chris Holp ATC Materials Lab holpcr-at-earthlink.net
Several people have already beat me to the punch, as they say, but I thoroughly agree with the rudeness of anonymity, even if it is unintentional. I have corresponded off-line with Nestor on this and my recollection is that the rules of the list preclude anonymous postings. Any one of us can occasionally make this error. But a small but annoying percentage of the members still post anonymously every time.
So why must I endure postings on asbestos when my interest is cathodoluminescence? Because it is part of the purpose of the list. And why must I endure questions from novices? Because this has also grown to be a purpose of the list and one that has been rather well served. And frankly it occasionally empowers me to ask questions that may seem stupid to those who know all things.
} Look in the phone book. } } Why must we endure these mundane questions, can anybody in this } world think for themselves. }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to dmrelion-at-aol.com. Thank you.)
At 08:52 PM 2/19/01 -0600, Gordon Couger wrote: } } From: {"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com} } } Look in the phone book. } } } } Why must we endure these mundane questions, can anybody in this } } world think for themselves. } } Not all of us live in a city, state or even a country that has a } microscope repair and service company.
Speak with respect and a humble aspect, for he is WizardOfTheLab ... -at-aol.com ! :-)
Although you were afraid to use your real name, and instead used an absurd pseudonym, allow me to enlighten you just the same. The asker of the question you have objected to was looking for a RECOMMENDATION. Facilitating the sharing of facts and opinions in a professional, helpful way is what this listserver does so well.
Perhaps you should consider unsubscribing?
Thanks, Jim
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- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - James F. Sanzo, Ph.D Dept. of Bioengineering 120 Hayden Hall 3320 Smith Walk University of Pennsylvania Philadelphia, PA 19104-6392
} -----Original Message----- } From: "Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com] } Sent: Monday, February 19, 2001 5:53 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Cleaning and maintenance of microscopes } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Look in the phone book. } } Why must we endure these mundane questions, can anybody in } this world think } for themselves. }
I am sorry you all are taking this so terrible, I was not intending to be rude. I was merely pointing out the fact that every time someone has a slight issue they want to magnify it in this forum. I think this forum can be used for better issues than this.
It seems to me that we are all intelligent people that can find things out for ourselves. We know enough to log onto the Internet and check what is in this forum everyday. I don't see why a simple Internet search for this service cannot be performed on any search engine. I performed a search on Yahoo and found 1000+ results. I narrowed it down to an area and found 16 in my area. We coddle people in this country like they are helpless children, we need to stop this or everybody will expect us to do everything for them.
Now I ask you do we really need this type of question in this forum?
If you all feel this is still rude I will unsubscribe.
} Now I ask you do we really need this type of question in this forum? } *** Yes. When I move to a new area, I ask colleagues to recommend a dentist; I don't merely go to the yellow pages or the web and hope for the best. In a similar vein, for most of us, our scopes are too precious to let just anyone touch them; a recommendation from a satisfied colleague is most valuable.
However, it would have been useful for the original poster to let us know where he/she was located. People from across the world subscribe to this list. A cleaning service in New Jersey may not be useful to the person in California or Australia.
Don ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
Try Fisher Scientific or ask a nearby university, department of biology. Good luck.
} } } {"Luvstodance99-at-aol.com"-at-sparc5.microscopy.com} 02/19 1:07 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are searching for companies that provide cleaning and maintenance of microscopes.
These kinds of services are not in fact so easy to locate. Our yellow pages would tell you nothing useful and web searches do not provide a full list or carry any personal recommendation. The question was simple, the reply can be equally simple or you can simply delete and ignore it. The usefulness of a list like this lies in the fact that even a simple question stands a high chance of being quickly and appropriately answered, because the users have a broad range interests and experience in common. Let's not diminish this by inhibiting people from participating.
Chris
ps - who and where are you anyway?
} } I am sorry you all are taking this so terrible, I was not intending to be } rude. I was merely pointing out the fact that every time someone has a slight } issue they want to magnify it in this forum. I think this forum can be used } for better issues than this. } } It seems to me that we are all intelligent people that can find things out } for ourselves. We know enough to log onto the Internet and check what is in } this forum everyday. I don't see why a simple Internet search for this } service cannot be performed on any search engine. I performed a search on } Yahoo and found 1000+ results. I narrowed it down to an area and found 16 in } my area. We coddle people in this country like they are helpless children, we } need to stop this or everybody will expect us to do everything for them. } } Now I ask you do we really need this type of question in this forum? } } If you all feel this is still rude I will unsubscribe. }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Yes, it is still rude. And it still doesn't address the issue of people who live and work outside of major centres. And you are still anonymous.
Lesley Weston.
On Tue, 20 Feb 2001 Wizardofthelab-at-aol.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am sorry you all are taking this so terrible, I was not intending to be } rude. I was merely pointing out the fact that every time someone has a slight } issue they want to magnify it in this forum. I think this forum can be used } for better issues than this. } } It seems to me that we are all intelligent people that can find things out } for ourselves. We know enough to log onto the Internet and check what is in } this forum everyday. I don't see why a simple Internet search for this } service cannot be performed on any search engine. I performed a search on } Yahoo and found 1000+ results. I narrowed it down to an area and found 16 in } my area. We coddle people in this country like they are helpless children, we } need to stop this or everybody will expect us to do everything for them. } } Now I ask you do we really need this type of question in this forum? } } If you all feel this is still rude I will unsubscribe. } }
} It seems to me that we are all intelligent people that can find things out } for ourselves. We know enough to log onto the Internet and check what is in } this forum everyday. I don't see why a simple Internet search for this } service cannot be performed on any search engine. I performed a search on } Yahoo and found 1000+ results. I narrowed it down to an area and found 16 in } my area. We coddle people in this country like they are helpless children, we } need to stop this or everybody will expect us to do everything for them. } } Now I ask you do we really need this type of question in this forum?
Perhaps you can enlighten some of the rest of us on the ease of searching. I checked the Yahoo yellow pages for "microscope cleaning" and found nothing in Iowa and not even anything around Chicago. I tried the regular Yahoo and Google searches and found thousands of matches, but I was not looking for a "coin cleaning microscope" or "cleaning a microscope slide". In view of such irrelevant results, I am not surprised that someone would ask the list for recommendations.
Perhaps you could give us a tutorial on how you were able to refine the search to produce more useful results. I consider myself rather web savvy and still fight the search engines. Such a tutorial could be a contribution rather than so much noise.
I did find the service we use for cleaning our LMs in our local yellow pages. It turns out to be through the distributor who first sold us the scopes. However, if I were not in a university town, I doubt that there would have been a listing.
} If you all feel this is still rude I will unsubscribe.
I don't know about rude, but I still have no idea of who you are. I don't know what credentials or experience you might have that I should give any consideration to your comments. An identity would help.
And in case anyone does not recognize iastate.edu as the domain for Iowa State University...
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Chris: have you tried the ASM International website? (http://asminternational.org) Try the "Shop ASM" button at the top right to see their library of book, videos, etc. I'm looking at one now called "Plastics Failure Guide: Causes and Prevention" by Myer Ezrin. They also have lots of information on ceramics. You can also search for information. I'm a member of one of their affiliate societies, the Electronic Devices Failure Analysis Society. (You don't have to be a member to but from them, although it is cheaper.)
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I don't think it would hurt if you identified yourself. Your mask of anonymity isn't helping matters much.
Gail Harrison Reichhold
-----Original Message----- } From: "Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com [mailto:"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, February 20, 2001 11:26 AM To: microscopy-at-sparc5.microscopy.com
I am sorry you all are taking this so terrible, I was not intending to be rude. I was merely pointing out the fact that every time someone has a slight issue they want to magnify it in this forum. I think this forum can be used for better issues than this.
It seems to me that we are all intelligent people that can find things out for ourselves. We know enough to log onto the Internet and check what is in this forum everyday. I don't see why a simple Internet search for this service cannot be performed on any search engine. I performed a search on Yahoo and found 1000+ results. I narrowed it down to an area and found 16 in
my area. We coddle people in this country like they are helpless children, we need to stop this or everybody will expect us to do everything for them.
Now I ask you do we really need this type of question in this forum?
If you all feel this is still rude I will unsubscribe.
Since you asked, yes, you were rude. No need to unsubscribe, though. Just try to show common courtesy. If you don't care about a given question, then don't reply. If you're interested in issues of self-reliant individuals and non-coddling societies, you might take your own advice and do a simple Internet search for an appropriate forum.
Vachik Hacopian
} I am sorry you all are taking this so terrible, I was not intending to be } rude. I was merely pointing out the fact that every time someone has a } slight } issue they want to magnify it in this forum. I think this forum can be } used } for better issues than this. } } It seems to me that we are all intelligent people that can find things out } for ourselves. We know enough to log onto the Internet and check what is } in } this forum everyday. I don't see why a simple Internet search for this } service cannot be performed on any search engine. I performed a search on } Yahoo and found 1000+ results. I narrowed it down to an area and found 16 } in } my area. We coddle people in this country like they are helpless children, } we } need to stop this or everybody will expect us to do everything for them. } } Now I ask you do we really need this type of question in this forum? } } If you all feel this is still rude I will unsubscribe.
Rude language and anonymity are not appropriate at all. Another point of view: ListServer is our public place for questions, answers and some "social talking". This space we have to keep clean. If for some reason you (or somebody else) don't like this place as it is- you may find many spaces on the Internet, where your language and anonymity is welcomed. Just go to the yellow pages...
} Date: Tue, 20 Feb 2001 11:25:34 -0500 (EST) } From: "Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com } Subject: Re: Cleaning and maintenance of microscopes } To: microscopy-at-sparc5.microscopy.com } X-Mailer: AOL 5.0 for Windows sub 117 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Looks like bad timing for this question on the list, but the SEM won't wait.
Would appreciate leads on third party maintenance providers for an Amray 1830 SEM located in Sacramento, CA. This system has a Balzers 240 turbo, LaB6 gun, Varian 30L/s ion pump, load lock system, and is currently on-line and working.
Providers of service contracts for this system are sought. System is at a DOD facility.
I think this kind of questions should always be welcome. The list serve is here to entertain all kinds of microscopy related questions no matter how simple it is. I posted a similar question last year and received some excellent replies. I don't think yellow pages or internet search would have done it. We are located in a small city and I can't imagine the yellow pages will have any kind of microscopy services.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
The whole idea of this listserver is to synergistically combine the expertise of people from a variety of backgrounds, locations, and experience levels so we can assist each other. I suggest you rejoin those of us who (usually) quietly lurk until you realize that flaming is neither helpful nor appropriate.
The person who requested assistance was thinking for herself - and doing it well. After all microscopists are the best source of information on individuals/companies truly qualified to clean microscopes. I do standard cleaning/maintenance on my PLM's and stereomicroscopes at work and have cleaned up many low end 'scopes as a freebie for public schools. But I wouldn't dream of fully disassembling/cleaning some of the high end microscope systems I've used, even if I had the tools available. I'd want a true expert - not just a name I pulled from the yellow pages or internet.
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-339-7300 phone 508-339-4996 fax Louise_Harner-at-albint.com
"Wizardofthela b-at-aol.com" To: microscopy-at-sparc5.microscopy.com cc: 2001/02/19 Subject: Re: Cleaning and maintenance of microscopes 06:52 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Look in the phone book.
Why must we endure these mundane questions, can anybody in this world think for themselves.
I'm an engineer in Nikon Japan. I used to work as sales engineer in NY. How old and which maker's microscope do you mention? If it is Nikon, could you contact our service or dealer? I know our service department clean and maintain microscopes with reasonable charge. Please contact 516-845-7788 first.
Hisashi Okugawa 1st design department Instrument company in Nikon Corporation Phone: 81-45-853-8568 Fax: 81-45-853-8475 e-mail: okugawa.h-at-nikon.co.jp ----- Original Message ----- } From: {"Luvstodance99-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 20, 2001 3:07 AM
No Great Loss.
Earl
----- Original Message ----- } From: {"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 20, 2001 2:08 PM
I am hesitant to reply to such a message, a few list members have expressed much of my sentiments, but think that there is a deeper problem with a few (very few but vocal) members of this list. I realize I am preaching to the choir and those that should hear, won't. Attacks such as this need to be discouraged.
Because of the quickness of a very few to attack without provocation, usually my answers are restricted to off the list responses. This gets the information to the inquirer, without requiring e-mail after e-mail in some sort of battle. There have been useful lively conversations that I learned much from, and gained deeper understanding, but these have always remained civil, and dealt with the question at hand. Responses such as below, really stifle I want from the list as a list member, a free flow of questions and answers. These are what I learn from. There is a delete key or trash bin for what I don't want to read, doesn't take long, nor much effort. I have an enormous file of those I keep for future reference and things I didn't know. The list works!
This list is for questions, hopefully for which, the inquirer doesn't readily have the answer. What is to one, mundane and trivial, to others is a significant question. Who is to chose what is mundane and for that matter what is the scope of the inquiry? When read, I understood that advice was being sought for the quality of service as well as who. This is the kind of information that is available through this list. I am sorry I wasn't able to respond to the original question but it was not from my region, so I can't suggest good service engineers in the inquirer's area to help.
I for one, would NOT want to trust a research light microscope worth $30,000.00 or more to a choice from the Yellow pages (nor the Web for that matter) without further information. We all have thrown bad service personnel out of the lab, and had to pay the price of having the job done right later. There is no information in the Yellow pages on how good or how bad a given company is. Users of these services DO have this information and we are the users.
As you send an e-mail, ask yourself, if the response you are giving doesn't answer, or some way relate to, the question, why are you sending it, and what good will it do for fellow list members?
Remember: There are no stupid questions, only stupid answers.
At 06:52 PM 2/19/2001 EST, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Well, I thought I'd see if I, too, could find 1000+ microscope maintenance companies in the US, and there are a few out there. Search under microscope maintenance:
www.southernmicro.com in Georgia southernmicroscope.com/services.html somewhere in the South www.dscoptical.com/services.htm near Boston www.caleywhitmore.com near Boston www.opticalexpertise.com in New York www.mwrn.com/product/microscope/repair.htm lists various service companies www.sciscope.com/field-service.htm based in Iowa, service most states www.svms.com/services/index.html in silicon valley www.meyerinst.com/html/mts/mts.htm in Texas www.microresource.com/services2.htm in Illinois www.allometrics.com/microscope_serv.htm in Lousiana and Texas www.labworksinc.com in San Fransisco www.biotherm-inc.com in 5 southern states www.mikroni.com/fp/fp/services.htm in San Diego www.uams.edu/oas/OES/oesilab.htm in Arkansas www.uni.edu/pur/maintenance/equipment.html Uni of Northern Iowa www.dominionmicroscope.com/services.htm Richmond, VA
etc.
took an hour to weed through them, while staining some sections......
cheers,
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
"I would like to know if anyone can recommend any books regarding the failure modes and mechanisms of plastics and ceramics. My searches have been yielded little (read as nothing), and my library is woefully lacking.
Thanks to all in advance for your help!"
Chris Holp ATC Materials Lab holpcr-at-earthlink.net
I have been working with SEM since the beginning of the 80's and one of my literature companions during all these years has been the excellent book "An Atlas of Polymer Damage", ISBN 0 7234 0750 9. Regarding ceramics I have little experience and do not know any good source.
Best regards
Pierre Frykberg Alfa Laval Tumba AB Div. Materials
Relax, and welcome to a public (though moderated) forum.
You got flamed because you failed to realize the extent of this list. I've been a denizen for many years and have found questions and responses on every level of expertise. As an internet resource, this list is exceptional; however, you have to be responsible for your own filtering as to what responses you make.
Basically, those posting here are interested primarily in constructive information. Had you requested the original poster's location and provided them with the results of your searches located within their area, there would have been nothing left to argue. You would have been lauded as helpful and the poster would have had the help they requested.
No whiney children here, only people of a wide variety of different abilities and experience in this narrow field of knowledge who hope to find some deeper understanding and practical help in fulfilling the tasks they have to pursue each day.
I'm sorry if you really do leave this list. Having been in this field for over 20 years, I rather enjoy the infusion of newcomers.
As an independent businessman, I am a staunch republican and arch-rival of 'political correctness', but I see none in this thread. A simple question was asked, requiring a simple answer. If you didn't have that simple answer, you should have just remained silent and deleted the email. Instead, you chose to inject your displeasure that this particular email was sent. Here's a surprise - this list is not maintained for your pleasure, but for the practical use of hundreds, if not thousands, of others.
You are welcome here, at least by me (I certainly can't speak for anyone else here). Please recognize that you are preaching to the masses here, and not just to an audience that you desire.
On Tuesday, February 20, 2001 4:08 PM, "Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com [SMTP:"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com] wrote: } } I am going to make you all happy and unsubscribe. You are making this exactly } what I was hoping to avoid. } } You all sound like a bunch of whiny children. } } Political correctness is ruining the fiber of this great country. } } Enjoy your pathetic lives. } } Mike Nolan } President } Materialographic Technologies } } P.S. I am using a back-up ISP while my Primary ISP is working out some } problems and I did not realize my signature was not included. } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
With regard to the recent "point / counter point" thread regarding the rude postings of "the Wiz", I was initially reluctant to "add to the pile" when so many had already voiced many of my sentiments. For years, I have encouraged my SEM students at a local university to subscribe to "the List" and regularly read and learn from the problems and questions many people have shared with List members. If the students had specific questions or similar problems, I have further encouraged them to post their comments or questions. The most common response from students is "I don't DARE post my silly question because everyone else will know the answer and I will look foolish. I don't want to be put down or scolded before thousands of knowledgeable readers" (I might add, who does?!) Until this recent thread from "the Wiz", I could comfortably assure my students the list was a place for good information without the assault of self appointed experts like "the Wiz". The true damage a rude person like "the Wiz" does is to discourage others from responding eventually leading to the death of the list server.
Problems like this one are inevitable. A few years ago, a forensic list server was plagued with an even more blatant and as you would guess, anonymous flamer. The flamer took delight in ridiculing many list members and seeing how far he/she/they could upset the list members. The list response was much the same as we are experiencing with "the Wiz". Our words will not beat him/her/they back and their offer to unsubscribe is a joke. Like a bully on a playground, if everyone ignores him/her, they will loose interest and go away. If he/she/they cannot master to ability to delete unwanted questions or text, how can we possibly expect them to abide by the unpublished rules of good manners and mutual respect?
When I have learned EVERYTHING about ALL types of microscopy, I may see "the Wizard's" view. But until then, I will enjoy and learn from fellow microscopists and their responses to the MSA List server. Let's not waste more personal (or company) resources on "the Wiz".
Keep up the good work, Nestor!
OBLIGATORY DISCLAIMER: My comments and opinions expressed in the above text are mine alone and do not represent those of the US FDA or any other Federal agency.
S. Frank Platek Forensic Chemistry Center U.S. Food and Drug Administration 6751 Steger Drive Cincinnati, OH 45237-3097 (513) 679-2700 X254 fplatek-at-ora.fda.gov
} From: "Allen R. Sampson" {ars-at-sem.com} Send reply to: "ars-at-sem.com" {ars-at-sem.com} To: "'\"Wizardofthelab-at-aol.com\"-at-sparc5.microscopy.com'" {"Wizardofthelab-at-aol.com"-at-sparc5.microscopy.com} , "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com}
Hi Listers,
Nestor, would you please call a halt to all the flying insults?
Everybody is getting their panties in a knot over this and I think it should end. Wizard was rude, but the replies are rude to.
To quote a "friend" of the LAPD ;-), "Can't we all just get along?"
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
More years ago than I want to mention, whenever we changed the filament on a Siemens 102 we cleaner the ceramic insulator with some sort of polish to remove any C buildup. Does anyone have a suggestion what to use as a "polish" for this.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id JAA13196 for dist-Microscopy; Wed, 21 Feb 2001 09:16:46 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id JAA13192 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 21 Feb 2001 09:16:16 -0600 (CST) Received: from e1g1.home.nyu.edu (E1G0.HOME.NYU.EDU [128.122.108.150]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id JAA13181 for {microscopy-at-sparc5.microscopy.com} ; Wed, 21 Feb 2001 09:16:05 -0600 (CST) Received: from kal-s-quantex (mcs01-ext.med.nyu.edu [128.122.2.2]) by e1g1.home.nyu.edu (8.10.1/8.10.1) with ESMTP id f1LFEJt20359 for {microscopy-at-sparc5.microscopy.com} ; Wed, 21 Feb 2001 10:14:19 -0500 (EST) Message-Id: {4.2.0.58.20010221101117.009544a0-at-imap.nyu.edu} X-Sender: kr4-at-imap.nyu.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 4.2.0.58
} I'm not sure about this, but I recall someone telling me that the current } Leica eyepieces for the MZ series of stereomicroscopes will also work on the } older Wild M5, M8, etc.
Now that a little of the invective has quieted down, here's the contact information for John Sowers, who services Amenex's light microscopes from his location in Claymont, Delaware:
J&L Microscope Services, (302) 798-5304
John has been servicing our B&L and Olympus microscopes for about fifteen years; they still work as well or better than they did when he started. What more can I say ?
Tell John I put you on to him.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
If it is a glazed insulator then we find that alumina (Al2O3) powder mixed with alcohol to form a slurry is good. Dipping a cotton bud in the slurry and working at the mark, then a general clean all around the component with the slurry on wipes is very effective. The alumina is easy to clean off and does not leave any residue. It is very important to wash thoroughly with frequent changes of alcohol.
If the tracking is very bad (deep) then it may be necessary to shot blast with alumina and then wash in alcohol. Shot blasting also works for some of the porous insulators.
Good luck, Ron
On Wed, 21 Feb 2001 07:27:08 -0600 (CST) "L. D. Marks" {ldm-at-risc4.numis.nwu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } More years ago than I want to mention, whenever we changed } the filament on a Siemens 102 we cleaner the ceramic insulator } with some sort of polish to remove any C buildup. Does anyone } have a suggestion what to use as a "polish" for this. } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
The bottom line is that "The List" should not be blamed for the opinion of "The Wiz". Here in America we advocate free speech, and we should be used to the idea that unpopular opinions will therefore occasionally be expressed -- and we shouldn't overreact to them. "The List" is an open forum, and so "The Wiz" has a perfect right to express his opinion. But students and beginners should realize that most of us don't share that opinion in this case, and they should feel perfectly free to ask questions about anything they find confusing in the vast realm of microscopy. It doesn't make any sense to me that anyone would abandon ship (unsuscribe) just because an unpopular opinion was expressed. We need collectively to have "thicker skins".
Kent Christensen
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Office: 5801 Medical Science II Building Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 E-mail: akc-at-umich.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Wed, Feb 21, 2001 6:38 AM -0500 "Platek, Frank" {FPLATEK-at-ora.fda.gov} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } With regard to the recent "point / counter point" thread regarding the } rude postings of "the Wiz", I was initially reluctant to "add to the } pile" when so many had already voiced many of my sentiments. For years, } I have encouraged my SEM students at a local university to subscribe to } "the List" and regularly read and learn from the problems and questions } many people have shared with List members. If the students had specific } questions or similar problems, I have further encouraged them to post } their comments or questions. The most common response from students is } "I don't DARE post my silly question because everyone else will know the } answer and I will look foolish. I don't want to be put down or scolded } before thousands of knowledgeable readers" (I might add, who does?!) } Until this recent thread from "the Wiz", I could comfortably assure my } students the list was a place for good information without the assault of } self appointed experts like "the Wiz". The true damage a rude person } like "the Wiz" does is to discourage others from responding eventually } leading to the death of the list server. } } Problems like this one are inevitable. A few years ago, a forensic list } server was plagued with an even more blatant and as you would guess, } anonymous flamer. The flamer took delight in ridiculing many list members } and seeing how far he/she/they could upset the list members. The list } response was much the same as we are experiencing with "the Wiz". Our } words will not beat him/her/they back and their offer to unsubscribe is a } joke. Like a bully on a playground, if everyone ignores him/her, they } will loose interest and go away. If he/she/they cannot master to ability } to delete unwanted questions or text, how can we possibly expect them to } abide by the unpublished rules of good manners and mutual respect? } } When I have learned EVERYTHING about ALL types of microscopy, I may see } "the Wizard's" view. But until then, I will enjoy and learn from fellow } microscopists and their responses to the MSA List server. Let's not waste } more personal (or company) resources on "the Wiz". } } Keep up the good work, Nestor! } } OBLIGATORY DISCLAIMER: } My comments and opinions expressed in the above text are mine alone and do } not represent those of the US FDA or any other Federal agency. } } } S. Frank Platek } Forensic Chemistry Center } U.S. Food and Drug Administration } 6751 Steger Drive } Cincinnati, OH 45237-3097 } (513) 679-2700 X254 } fplatek-at-ora.fda.gov }
At 03:03 AM 2/21/01, you wrote: } I don't know if it will help, but here's the response I had to make earlier } today to a DOD request in the Sacramento area. The email address I was } given didn't work, so I will have to call the party tomorrow. He claimed } that they had three Amrays at their location. Amray's new service policies } have certainly created some problems for customers!
Are any of you aware of a change in Amray's (KLA-Tencor) service policies? It seems that their main problem is lack of trained personnel. The second driver is the diminishing of importance of the lab SEMs versus the KLA semiconductor fab/inspection equipment.
The problem with this particular 1830 system is that pre-contract inspection is done on a per-diem basis. This is the lowest priority service call. Systems are not placed on contract until the pre-inspection is done. Sort of a Catch-22 situation.
This may not be a huge problem for systems like a 1000 or 1600 or 1610T. Maybe not too bad for an 1830. Non-Amray service ought to be fine. But for FESEMs like 1840, 1845, 1880, 1910, 3300 and 3600, it is quite another matter. Is there an impression or actual set of data points where Amray/KLA is dropping the non-FESEMs and retaining the FESEMs or simply dropping all models of SEMs?
KLA-Tencor, Amray Division recently sent me a notice they were no longer providing service contracts for my area (Michigan). They also sent a list of eleven third party service providers. We contracted with one of those vendors for our service (Ex-Amray guys of course). The vendor we chose simply accepted the instrument without inspection since we were just coming off contract with KLA.
I can't speak for FESEMs since ours is an 1820 LaB6.
Good luck with your search.
Kevin
--------------------------------------------------------------- Kevin Battjes Senior Research Specialist Michigan Molecular Institute Voice 517-832-5555, ext 556 1910 W. St Andrews Road Fax 517-832-5560 Midland MI 48640 e-mail: battjes-at-mmi.org After April 6, 2001 use 989 area code ---------------------------------------------------------------
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, February 21, 2001 11:37 AM To: ars-at-sem.com Cc: MSA listserver
At 03:03 AM 2/21/01, you wrote: } I don't know if it will help, but here's the response I had to make earlier } today to a DOD request in the Sacramento area. The email address I was } given didn't work, so I will have to call the party tomorrow. He claimed } that they had three Amrays at their location. Amray's new service policies } have certainly created some problems for customers!
Are any of you aware of a change in Amray's (KLA-Tencor) service policies? It seems that their main problem is lack of trained personnel. The second driver is the diminishing of importance of the lab SEMs versus the KLA semiconductor fab/inspection equipment.
The problem with this particular 1830 system is that pre-contract inspection is done on a per-diem basis. This is the lowest priority service call. Systems are not placed on contract until the pre-inspection is done. Sort of a Catch-22 situation.
This may not be a huge problem for systems like a 1000 or 1600 or 1610T. Maybe not too bad for an 1830. Non-Amray service ought to be fine. But for FESEMs like 1840, 1845, 1880, 1910, 3300 and 3600, it is quite another matter. Is there an impression or actual set of data points where Amray/KLA is dropping the non-FESEMs and retaining the FESEMs or simply dropping all models of SEMs?
To my knowledge KLA Tencor got out of the lab SEM business to include service. They did not renew our service contract and supplied a list of third party vendors . This is where I got Grant Gerringer's name.
I would suggest you contact AMRAY directly and get the list of third party vendors and a copy of the letter.
If it is a glazed insulator then we find that alumina (Al2O3) powder mixed with alcohol to form a slurry is good. Dipping a cotton bud in the slurry and working at the mark, then a general clean all around the component with the slurry on wipes is very effective. The alumina is easy to clean off and does not leave any residue. It is very important to wash thoroughly with frequent changes of alcohol.
If the tracking is very bad (deep) then it may be necessary to shot blast with alumina and then wash in alcohol. Shot blasting also works for some of the porous insulators.
Dear Laurence,
If the shot-blasting is necessary, you may want to check out the Air Eraser, available from Scientific Instrument Services. It's not expensive and can operate on valved-down house compressed air. It has a small tip, so it can easily be controlled.
Look at this dialogue as "theater-a la-internet" and we can see this in many different perspectives. Social commentary? Have we abdicated our ability to understand more than one perspective? It makes for great life art. I like to see it as an aggravation, a lesson, and humor....being in the audience...but then I love EM as art!
While we are expressing our feelings, let me strongly suggest that subscribers temporarily unsubscribe if they are going to use an autoanswer reply when out of the office.
If one posts to this list, one is addressing the entire list and a stream of autoanswer SPAM should not result. Frankly, my dears, I don't like getting autoanswer messages from people I do not know informing me how long they will be away.
There was an earlier posting requesting information and leads for x-ray inspection systems for integrated circuits. I must have lost the post but I have found a recent lead for what seems like a nice system. It is quoted in the $75K price range as was at the requested budget amount in the original posting.
I hate to be the bearer of bad news to the list but I feel many people will want to know this. After a long battle with cancer Terry Donovan passed away yesterday. While his primary association with electron microscopy was as a salesman he nonetheless had a vigorous enthusiasm for the field and promoted it where ever he went. No doubt that over the years he touched the lives of many people in a positive way, as he did mine. Anyone who knew he was a sailing enthusiast may be interested to know he wanted his ashes put into the Chesapeake Bay. Regards........Alan Berginc Cressington Scientific, Inc 508 Thomson Park Drive Cranberry Township, PA 16066-6425 TEL 724-772-0220 (USA)
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I am relatively new to this listserve and have been quietly monitoring the various threads and wide array of responses. Although much has no relevance to my work, I have found many responses quite useful and thank the list for the opportunities it provides.
I am passing along a request for recommendations on image analysis software. A client has a collection of fluorescence micrographs (film originals) that they would like to analyze quantitatively. They do not have a digital capture system that allows them to do the quantitation real time. I believe it is a simple matter of thresholding and measurimg the areas, but I may be missing something in my assumptions.
I have already recommended NIH (Scion) Image for the purpose, as I have used an early version of that software for similar purposes, but they would like to know if there are any highly recommended commercial packages that might be better, or if there are any companies that could do this analysis for them (again from their film originals).
The request comes from a graduate student at the University of Illinois in Chicago who has money for this purpose.
Any suggestions either through the list or to me personally are greatly appreciated and I will forward those responses on.
Thanks for your time.
James Hayden
*********************************
James E. Hayden, RBP, FBCA Bio-Graphics 1058 Hemlock Drive Blue Bell, PA 19422 tel/fax: (215)654-0625 tel:(215)514-4223
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At 02:55 PM 2/21/01, you wrote:
(snip) } This may not be a huge problem for systems like a 1000 or 1600 } or 1610T. Maybe not too bad for an 1830. Non-Amray service } ought to be fine. But for FESEMs like 1840, 1845, 1880, 1910, } 3300 and 3600, it is quite another matter. Is there an impression } or actual set of data points where Amray/KLA is dropping the non-FESEMs } and retaining the FESEMs or simply dropping all models of SEMs?
The official word from Amray is that they are supporting the FESEM at this time & will do so "for probably another five years". As I do not have that in writing, the "five year" policy may change.
} Are any of you aware of a change in Amray's (KLA-Tencor) service } policies? It seems that their main problem is lack of trained personnel. } The second driver is the diminishing of importance of the lab SEMs } versus the KLA semiconductor fab/inspection equipment. } } The problem with this particular 1830 system is that pre-contract inspection is } done on a per-diem basis. This is the lowest priority service call. } Systems are not placed on contract until the pre-inspection is done. } Sort of a Catch-22 situation.
I don't understand. Are you trying to obtain a service contract from Amray at this time? Or are you trying to obtain a contract from a "third party' maintenance organization?
Regards,
Earl
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: {ars-at-sem.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, February 21, 2001 8:37 AM
They do exist.
You might try "Aspex" (formerly RJ Lee Instruments) at (724) 744-0100.
Earl
----- Original Message ----- } From: "rad0" {rden25-at-mindspring.com} To: "microscopers!!" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, February 21, 2001 11:06 AM
I am fairly sure this is the case. I had a Leica MZ8 and MZ6 and the eyepieces could be interchanged with the Wild M8 I had. I am not sure about tube lengths and all that if such things are critical on stereo microscopes.
I have recently been approached by some of our researchers with the request to assist them in developing an imaging system that enables them to measure the thickness of a liquid and/or vapour film that develops when water is sprayed onto a hot metal surface.
In approaching me with their request they hoped that my experience in electron microcopy might give them some ideas. However, I am at a loss.
Can anyone out there give me advise whom to contact and what technique has been developed to measure (in situ) liquid film thicknesses in the order of approximately 30 microns.
Thanks
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
Optical techniques might work. Ellipsometry or other reflection techniques. Contact someone at JA Woollam in Lincoln, Nebraska.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Hans Brinkies [mailto:HBrinkies-at-groupwise.swin.edu.au] Sent: Wednesday, February 21, 2001 8:33 PM To: microscopy-at-sparc5.microscopy.com Subject: Liquid film thicknesses
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I have recently been approached by some of our researchers with the request to assist them in developing an imaging system that enables them to measure the thickness of a liquid and/or vapour film that develops when water is sprayed onto a hot metal surface.
In approaching me with their request they hoped that my experience in electron microcopy might give them some ideas. However, I am at a loss.
Can anyone out there give me advise whom to contact and what technique has been developed to measure (in situ) liquid film thicknesses in the order of approximately 30 microns.
Thanks
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
I've been getting a lot of inquiries from Amray customers for contract proposals, including one for a 3300 FESEM. In that case, I don't think that Amray made any gestures - they also have a 1600 series SEM and are not impressed with the service on the 3300 so we are talking about servicing both.
Don't be too alarmed. Some of us independents are the most experienced EM service personnel out there. I have over twenty years as an independent working on all EM manufacturers as well as a wide variety of other instruments such as XRF, AAS, UV-Vis, GC-MS, IR-FFT and a variety of microprobes, not to mention my previous experience with an SEM manufacturer, IBM and others.
The major concern for an FESEM owner transferring to an independent is, of course, the cathode assembly. Amray will charge $10,000 for a swap and billable installation on top of that. Denka, however, manufactures emitters that will work in most FESEMs for generally less than $3,000 and are available through Energy Beam Sciences. While there are increased demands in reconditioning the electrostatic surfaces in an FESEM cathode, they are easily managed. Given today's manufacturers service contract pricing, you should be able to find an independent coming in at nearly 50% less, not including the emitter replacements. Add in the purchase of one emitter per year, and your costs should still be considerably lower, while the experience level of your service engineer is increased.
Many FESEMs sold are located in companies that also have clean room facilities or laminar flow hoods. Clean rooms only certify a certain elimination of airborne particulates. While that is useful, it only addresses one part of the contamination concerns regarding FE cathodes - one that can be easily handled through a careful and methodical cathode assembly under nearly any conditions.
The other concern is that of chemical cleanliness of the lens surfaces. Once again, use of appropriate cleaning and handling methods can produce good results. There is nothing magical here, more than likely the manufacturers are doing the same work under conditions that may surprise you, not to mention the experience and qualifications of the employees doing the work.
Like a good jeweler or machinist, anyone experienced in servicing these machines should be able to eyeball dimensions down to 1/10,000 of an inch, perhaps with the help of a loupe. Alignment of any critical components shouldn't be any problem.
Don't be shy to ask the questions you have of any independent you may be considering. We survive on our integrity in a very limited field. If an independent can't properly handle customer questions and problems, they won't be able to feed their families for long. Believe me, it's tough to keep a business like this going if you can't at least equal the service capabilities of the manufacturer, as low a bar as that may be at times.
I can't tell you what Amray is considering other than the announcements they have already made. I can tell you that similar moves were considered by ISI prior to their withdrawal from the American market and subsequent marketing and service licensing through another manufacturer. While such a move is more than unlikely for Amray, I'd say that SEM manufacturers in general are looking for creative ways to get out of self-imposed difficulties.
It is possible that we are witnessing yet another new trend and other manufacturers may be considering similar moves. The SEM field has grown greatly over the past two decades, and manufacturers still seem unable to recognize the true value of their service organizations - that of marketing support by helping to build a base of loyal customers.
Instead, they are increasingly demanding their service organizations be independent profit centers. In doing so, they bring a bean-counter mentality to service that minimizes the concept of a career for service engineers. By doing so, they encourage a rapid turn-over in their service force which is why you probably have seen either a never ending stream of promising, but inexperienced, engineers or a civil-service stereotypical engineer who never quite makes the grade, but never leaves.
Sorry to have indulged in yet another of my tirades on the manufacturers. Now and then there appears one manufacturer who seems to get it, and they can profit greatly. There is one, who shall remain un-named, that I recommended to my customers for over a decade because of the post sales service they provided. I'm probably responsible for well over a million dollars a year in sales for them for five to ten years.
I saw the writing on the wall when they announced lay-offs at their corporate offices over five years ago; in a country where such a thing was unheard of at the time. Their response seems to have been a clamp down on expenditures and rising prices in foreign countries, witnessed by the increasing requests I've received for service on their instruments over the last few years. Frankly, I would have preferred to see them continue their monopoly on good service, as I only give advice I am reasonable sure of. However, having said that, I can state to everyone, as I stated in the advice that I have given in the past, that every manufacturer eventually f___s up.
Putting on the head dress of soothsayer, I can speculate that manufacturers are looking to increase their profits by dumping those lines that produce the least profit and concentrate on those lines that produce the most. Having everyone who has bought into the latest and greatest of technological advances, i.e. FE, snuckered into thinking that there are no alternatives to their magic, there are greater profits for the manufacturers in the service of these instruments. After all, the alternative would seem to be a nearly doubling of their already bloated service costs.
The trend over the last two decades has been to introduce new product models at a rapid rate, regardless of any real evolutionary improvements. These are due primarily to the increased influence of marketing departments that seek not just to distance themselves from the competition, but also from their own offerings. In this trend, manufacturers have screwed themselves as the learning curve for inexperienced service personnel for a large variety of models is much longer than is really necessary. The result is that they expect more from their service engineers and place increasing demands on them while not providing increased incentives. I've yet to see a service engineer promoted beyond service manager. Yet human resource, marketing and accounting underlings are commonly promoted to top corporate positions. Why stay in service?
I've seen the industry standard service contract cost rise from 5% of purchase price per year to over 10%. I have not seen any indication that those costs are justified, except through poor management decisions by the manufacturers. Since the purchase prices themselves reflect any basic cost increases over the years, exactly how do you justify the doubling of service costs which I've stated in proportion to the purchase price?
Have I given you a hint of where I'm coming from? Sorry, but sometimes when you give me a nail, I'll grab a 16 pound sledge hammer, although never when I'm working on an SEM. However, you may want to bolt your windows, as I often want to toss an instrument out. (To Fred, and anyone else who tends to take me too seriously, the immediately preceding was intended as a joke, as poor as it was).
On Wednesday, February 21, 2001 10:37 AM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } At 03:03 AM 2/21/01, you wrote: } } I don't know if it will help, but here's the response I had to make earlier } } today to a DOD request in the Sacramento area. The email address I was } } given didn't work, so I will have to call the party tomorrow. He claimed } } that they had three Amrays at their location. Amray's new service policies } } have certainly created some problems for customers! } } Are any of you aware of a change in Amray's (KLA-Tencor) service } policies? It seems that their main problem is lack of trained personnel. } The second driver is the diminishing of importance of the lab SEMs } versus the KLA semiconductor fab/inspection equipment. } } The problem with this particular 1830 system is that pre-contract inspection is } done on a per-diem basis. This is the lowest priority service call. } Systems are not placed on contract until the pre-inspection is done. } Sort of a Catch-22 situation. } } This may not be a huge problem for systems like a 1000 or 1600 } or 1610T. Maybe not too bad for an 1830. Non-Amray service } ought to be fine. But for FESEMs like 1840, 1845, 1880, 1910, } 3300 and 3600, it is quite another matter. Is there an impression } or actual set of data points where Amray/KLA is dropping the non-FESEMs } and retaining the FESEMs or simply dropping all models of SEMs? } } Thanks, } gary gaugler } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
There are quite a few ISI-40s still in use. As a basic SEM, it works. ISI made their inroads in the American market as a low bidder, offering a useable instrument at a price that others couldn't match.
There are a variety of independent service providers out there that can service your instrument, but you need to specify your location.
The last I knew, ISI made their schematics available at an incredibly low cost (I got mine at $35 US). Access to ISI is currently through Leo (have I got it right?) and you should contact their parts department for the schematics. Their schematics are in Japanese (or is it Korean?) so you should be cognizant of international electronic symbols and nomenclature. If not, then you should seek the help of someone who is. As far as I am aware, the electrical schematics are all that is available.
On Wednesday, February 21, 2001 1:06 PM, rad0 [SMTP:rden25-at-mindspring.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I have aquired the remnants of an ISI-40 sem. } } I'm new to this thing, and I'm going to need some sort } of technical manual just to identify the components. } } So, do these exist? Did they ever? } } And, if so, where might I start looking for them? } } And also, this thing looks mighty old, is this model still used } by anyone, and are there technicians still alive who could work } on it? } } Thanks, } Rick } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
I would like to announce the 1st International Meeting & Workshop on Advanced Light Microscopy - June 6-10, 2001 - Santa Maria Imbaro, Italy.
The program will include lectures on recent exciting advances in the fields of light microscopy and their applications to modern questions in life sciences. The practical workshop (afternoon), organised by manufacturers, leading in the field, will give the possibility to learn practical aspects on the latest developments in the field. Evening session will give students the possibility to discuss their own work and we also will have round table discussions bringing together manufacturers and scientists discussing where light microscopy presently stands and where it should move to in the future.
For further information, please check this web page: http://www.embl-heidelberg.de/ELMI/ItalyMeeting
Having dug further into my emails for the day, Earl has provided the right answer, contrary to my previous posting. I swear, its getting harder every day to track the passage of individual SEM manufacturers. RJ Lee did indeed handle the ISI assets in the US, my mistake, and you should contact them for info. The transfer to Aspex was not known to me, but web links to RJ Lee as a manufacturer appear to be non-existent and Earl is probably right. I'm confused...
On Wednesday, February 21, 2001 1:06 PM, rad0 [SMTP:rden25-at-mindspring.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I have aquired the remnants of an ISI-40 sem. } } I'm new to this thing, and I'm going to need some sort } of technical manual just to identify the components. } } So, do these exist? Did they ever? } } And, if so, where might I start looking for them? } } And also, this thing looks mighty old, is this model still used } by anyone, and are there technicians still alive who could work } on it? } } Thanks, } Rick } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Thank you all for the myriad of responses to the inquiry regarding failures in plastics and ceramics. Below is a brief compilation of suggested sources:
"Medical Plastics: Degradation Resistance & Failure Analysis", Portnoy, ISBN 1-884207-60-X "Plastics Failure Guide: Cause & Prevention", Ezrin, ISBN 1-56990-184-8 (These two are available at the www.asm-intl.org site)
"An Atlas of Polymer Damage" ISBN 0-7234-0750-9 "Failure of Plastics", Brostow & Corneliussen, ISBN 3-446-14199-5 "Polymer Microscopy" ISBN 0-412-25710-6 "An Atlas of Polymer Damage", Engle et al, ISBN 0-13-050013-5 "Polymer Degradation", W. Schnabel "Case Studies of Plastics Design & Failure Analysis", Mallick/Ford Motor Co. "Handbook of Plastics Technology", SPE "Polymer Characterization & Analysis", Kroschwitz "Handbook of Plastics Flammability", Landrock "Handbook of Plastics Degradation", Mamid & Amin
This is a quick list and I have not yet investigated each title, but I do have a few on order. I was actually impressed at the number of relevant to semi-relevant books shown at Amazon.com. Also, The Society of Plastics Engineers has a nice site with good book offerings.
Chris Holp ATC Materials Lab Cleveland, OH holpcr-at-earthlink.net
Access is through Aspex. Leo is a consortium of Zeiss & Cambridge.
----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'rad0'" {rden25-at-mindspring.com} ; "microscopers!!" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 22, 2001 1:42 AM
Dear Laurence, I have used six micron diamond polishing paste for this, in the past. It must be very thoroughly washed off with clean ethanol or methanol. At 07:27 AM 2/21/01 -0600, you wrote: } More years ago than I want to mention, whenever we changed } the filament on a Siemens 102 we cleaner the ceramic insulator } with some sort of polish to remove any C buildup. Does anyone } have a suggestion what to use as a "polish" for this. } } ------------------------------------------------------- } Laurence Marks Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I'm having a stabiliy problem with LR White sections. They curl, expand, split, etc. during examination on the TEM. My operating conditions are 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity low. In the past I've tried vac. evap. a layer of carbon which helped some. I was also thinking of coating the grids with another layer of formvar to "sandwich" the sections. I would appreciate anyone's thoughts and what they have tried in solving the stability problem. thanks.
We've had nothing but trouble with LR White and have finally switched to Unicryl to see if that improves things. Not only has LRW been unstable under the beam, but we've had repeated problems with sections being destroyed in the immunolabeling and staining processes. We have cleaned and recleaned our grids. We have tried different hardness grades of LRW and different batches of resin. We've tried extended dehydrations and infiltrations and abbreviated ones, from the manufacturer's suggested protocol to every sensible variation we could think of. The final straw was when we processed LRW and Unicryl embedded samples side by side and got beautiful stable sections in Unicryl and nonexistent to tattered shreds on the LRW grids.
We're still pretty new to Unicryl, so I'm sure it will have its own set of problems, but so far it's been more than acceptable.
There are some archived discussions about LR resins at http://www.biotech.ufl.edu/~emcl/tips.html.
Meanwhile, I'd be interested in seeing any responses you get, if you wouldn't mind.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Tom W Bargar [mailto:tbargar-at-unmc.edu] Sent: Thursday, February 22, 2001 1:12 PM To: Microscopy-at-sparc5.microscopy.com
I'm having a stabiliy problem with LR White sections. They curl, expand, split, etc. during examination on the TEM. My operating conditions are 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity low. In the past I've tried vac. evap. a layer of carbon which helped some. I was also thinking of coating the grids with another layer of formvar to "sandwich" the sections. I would appreciate anyone's thoughts and what they have tried in solving the stability problem. thanks.
I'm trying to track down a formulation for Teorell and Stenhagen buffer. A 2.5mM concentration of diaminobenzidine is used with this buffer and hydrogen peroxide to selectively stain the catalase in peroxisomes. The references that I have found all seem to use this buffer and reference an article in a German journal dating to 1938. Is anyone out there familiar with this buffer? If so, can you supp;y the recipe? Is there another buffer that I can use to effectively stain peroxisomes? As always, thanks in advance for your replies.
Tom Januszewski Senior Electron Microscopist UT Southwestern Medical Center at Dallas Dallas, TX 75390-9039 214-648-7291 FAX: 214-648-6408 Email: tom.januszewski-at-UTSouthwestern.edu
Tom - Have you tried sort of "burning" the sections at low mag, low beam for a few minutes when they first go in the beam? That may help stabilize them for work at higher mags. That has helped me in the past.
However, I'm joining the Unicryl camp :) Never had trouble with losing LR White sections during processing, but the few times I've used Unicryl so far I've had less trouble with microholes in the sections (had those in LR Gold, too) and the sections have been more stable in the beam.
Not bashing LR resins - they've worked just fine for me for years, but I'm all for trying newer stuff when I can!
Tamara
On Thu, 22 Feb 2001, Tom W Bargar wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm having a stabiliy problem with LR White sections. They curl, expand, } split, etc. during examination on the TEM. My operating conditions are } 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity } low. In the past I've tried vac. evap. a layer of carbon which helped some. } I was also thinking of coating the grids with another layer of formvar to } "sandwich" the sections. I would appreciate anyone's thoughts and what they } have tried in solving the stability problem. thanks. } } Tom Bargar } EM Lab, UNMC } Omaha, NE } (402)559-7347 } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Tom, I use routinely formvar/carbon coated copper grids. The other way of support I use, especially for high resolution work and elemental analysis where I don't want to look through the layer of carbon, is a holey "lacey" carbon film. The support is superb, the drawback is a limited viewing area. I purchase both from EMS. I've never tried to make a sandwich with another formvar layer, but sounds to me kind of labor-intensive (not speaking of another layer you'd be imaging through). Good luck, Alice.
Alice Dohnalkova Battelle, PNNL MS P7-50 Richland, WA 99352 (509) 372-0692
-----Original Message----- } From: Tom W Bargar [mailto:tbargar-at-unmc.edu] Sent: Thursday, February 22, 2001 11:12 AM To: Microscopy-at-sparc5.microscopy.com
I'm having a stabiliy problem with LR White sections. They curl, expand, split, etc. during examination on the TEM. My operating conditions are 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity low. In the past I've tried vac. evap. a layer of carbon which helped some. I was also thinking of coating the grids with another layer of formvar to "sandwich" the sections. I would appreciate anyone's thoughts and what they have tried in solving the stability problem. thanks.
RJ Lee Group had spun off RJ Lee Instruments a couple of years ago. RJ Lee Instruments, Limited changed it's name to Aspex, LLC last year. ASPEX stands for Application Specific Products using Electron beam and X-ray analytical technologioes, they tell me. It was easier for me to remember RJ Lee. For more info their web address is www.aspexllc.com . Which has some contact information or a button for questions as I recall. If ASPEX doesn't handle what you want I'm sure they can give you contact information for RJ Lee Group.
Jim Roberts
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } "Allen R. Sampson" {ars-at-sem.com} 02/22/01 02:13AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Having dug further into my emails for the day, Earl has provided the right answer, contrary to my previous posting. I swear, its getting harder every day to track the passage of individual SEM manufacturers. RJ Lee did indeed handle the ISI assets in the US, my mistake, and you should contact them for info. The transfer to Aspex was not known to me, but web links to RJ Lee as a manufacturer appear to be non-existent and Earl is probably right. I'm confused...
On Wednesday, February 21, 2001 1:06 PM, rad0 [SMTP:rden25-at-mindspring.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I have aquired the remnants of an ISI-40 sem. } } I'm new to this thing, and I'm going to need some sort } of technical manual just to identify the components. } } So, do these exist? Did they ever? } } And, if so, where might I start looking for them? } } And also, this thing looks mighty old, is this model still used } by anyone, and are there technicians still alive who could work } on it? } } Thanks, } Rick } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Below is the result of your feedback form. It was submitted by (muller.bruce-at-columbus.co.za) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, February 22, 2001 at 06:28:29 ---------------------------------------------------------------------------
Email: muller.bruce-at-columbus.co.za Name: Bruce Muller
State: Mpumalanga
Zip: 1050
Question: What solution and method would one use to deep-etch Type 316 STAINLESS STEEL to reveal the inclusions and/or precipitates present in relief. This will be used for SEM analysis. Thankyou in advance for your assistance. Bruce
Hi I think there are a few things you should be careful with. First, don't use naked grids with LRW sections, always use film supported grids. Second, don't try to use chloroform to extend the wrinkles of your sections before picking them up. Third, don't use UAc in EtOH or MtOH, use aquatic UAc for your staining.
Good luck
Greg Ning EM Facility Medical College of Wisconsin
Tom W Bargar wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm having a stabiliy problem with LR White sections. They curl, expand, } split, etc. during examination on the TEM. My operating conditions are } 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity } low. In the past I've tried vac. evap. a layer of carbon which helped some. } I was also thinking of coating the grids with another layer of formvar to } "sandwich" the sections. I would appreciate anyone's thoughts and what they } have tried in solving the stability problem. thanks. } } Tom Bargar } EM Lab, UNMC } Omaha, NE } (402)559-7347
Hello, Can someone please give me a source for free or inexpensive Si wafer rejects? I break them into small squares to use as SEM substrates for nano particles. Thank you, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
The company that now owns ISI is the Optical company TOPCON they have branches through out the world and locally Topcon still has "ISI" instruments available. Regards JVN
"Allen R. Sampson" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } There are quite a few ISI-40s still in use. As a basic SEM, it works. ISI } made their inroads in the American market as a low bidder, offering a } useable instrument at a price that others couldn't match. } } There are a variety of independent service providers out there that can } service your instrument, but you need to specify your location. } } The last I knew, ISI made their schematics available at an incredibly low } cost (I got mine at $35 US). Access to ISI is currently through Leo (have } I got it right?) and you should contact their parts department for the } schematics. Their schematics are in Japanese (or is it Korean?) so you } should be cognizant of international electronic symbols and nomenclature. } If not, then you should seek the help of someone who is. As far as I am } aware, the electrical schematics are all that is available. } } On Wednesday, February 21, 2001 1:06 PM, rad0 [SMTP:rden25-at-mindspring.com] } wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello, } } } } I have aquired the remnants of an ISI-40 sem. } } } } I'm new to this thing, and I'm going to need some sort } } of technical manual just to identify the components. } } } } So, do these exist? Did they ever? } } } } And, if so, where might I start looking for them? } } } } And also, this thing looks mighty old, is this model still used } } by anyone, and are there technicians still alive who could work } } on it? } } } } Thanks, } } Rick } } } } } } } } Allen R. Sampson, Owner } Advanced Research Systems } 317 North 4th. Street } St. Charles, Illinois 60174 } voice 630.513.7093 fax 630.513.7092
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
I would pick up sections on the carbon-plastic-coated grid and then put second layer of carbon over. In such "sandwich", extra couple of nanometers of carbon does not harm your resolution. Carbon should work perfectly to protect your sections. Secondly, I would try higher HT, like 80 kV.
Good luck, Sergey
At 11:12 AM 2/22/01 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
A few years ago I did quite a bit of immunogold labelling on LR White sections on uncoated 400 mesh copper thin-bar grids (washed thoroughly with acetone) at 80-100 kV and imaging at 40-100K. I usually "baked" the sections a bit at low mag before imaging at high mag - as suggested by Tamara Howard. They were rather unstable, but I would usually be able to count gold particles and/or take a photo before the section blew up. As you say, keeping the beam intensity low prolonged their life a bit, enough to get the information needed. So I didn't solve the problem, but just worked around it as much as possible. Also, I cut thickish sections - dark gold, expanding to pale gold with chloroform vapour waved over the knife bath. Longer polymerisation might help, but then you may lose antibody access to the tissue, if that's what you're interested in. I used medium or hard grade resin polymerised ~16 hours at 55 C. Over 4 years, we did have a couple of dud batches that didn't polymerise very well.
good luck, Rosemary White } } I'm having a stabiliy problem with LR White sections. They curl, expand, } split, etc. during examination on the TEM. My operating conditions are } 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity } low. In the past I've tried vac. evap. a layer of carbon which helped some. } I was also thinking of coating the grids with another layer of formvar to } "sandwich" the sections. I would appreciate anyone's thoughts and what they } have tried in solving the stability problem. thanks. } } Tom Bargar } EM Lab, UNMC } Omaha, NE } (402)559-7347
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
The objectives of our Leitz DURIMET Micro-Hardness tester was damaged during class experiments last year. Has anyone an idea where I can purchase a x10 (A 0.18 C HM25~) and a x40 (A 0.7 C HM6.3~) objective that suits a Leitz DURIMET.
Thanks
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
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Depending on what you call cheap, Ted Pella sells 4" wafers precut to either 5x7 or 5x5mm chips for $61.00 US.
Dale
On Thu, 22 Feb 2001, JIM ROMANOW wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } Can someone please give me a source for free or inexpensive Si wafer } rejects? I break them into small squares to use as SEM substrates for nano } particles. } Thank you, } Jim } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } Unit-2131 } Storrs, CT 06269-2131 } bsgphy3-at-uconnvm.uconn.edu } 860 486-2914 voice } 860 486-1936 fax } } }
Dale Shumaker G9 WBSB 725 N Wolfe Street Baltimore, MD 21205
Wilson Lab; Johns Hopkins University School of Medicine, Cell Biology and Anatomy 410-614-2654 410-955-4129 (fax)
Hi Tom! LR-White behaves sometimes a little bit "excentric" ... I would like to give you some hints drawn from my personal experience with LR-White:
-I use medium grade LR-W. That´s not because I tried other grades, but this works well with me
- The way of LRW polymerization is critical. If you do it in the heat, at 55-60°C, with or without vacuum, plastic gets very hard and sometimes brittle. Although, it´s easy to cut but the sections tend to be unstable in the beam, they suffer badly during immunoprocedures (sometimes only 1 out of 4 survives...), they are destroyed by alcoholic UA, contrast is sometimes awfully low and so on ...
- I polymerize LR-White in the cold at -20° by addition of 15microlitres of accelerator to 1ml of fresh resin which is in good-sealing microtubes (air prevents polymerization). What I get is not that good to cut because is softer than heat-polymerized LR-W, but it comes off the knife without wrinkles, is very stable during processing and in the beam at even 80kV. Contrast is sufficient, even if you only stain with (aquaeous) Uranyl and omit the lead. Immunolabeling works fine if you add 0.025 % Tween-20 to washing buffer and secondary gold-probe.
- Yes its true, the older this stuff gets even in your fridge, the poorer polymerization (especially by the use of the accelerator) will be. One year should be a batch´s life more or less.
- I use simply formvar coated nickle/copper grids. That is sufficient. If you have the facilities to coat the grids with carbon, it might work even better, everything is more robust during immunolabeling.
Why don´t you try it with the accelerator. Important: be quick as polymerization sets on immeadiatly, for that cool down the resin together with the samples (-20°C), don´t polymerize more than a handful of samples at one time. Use some kind of plastic microtubes (eppendorf, sarstedt and so on ...) which seal tight! After adding accelerator to the cold resin, shake vigorously and than hurry up. Make a paper copy of sample numbers and use them, laser-printouts get smeary and unreadable after LRW polymerziation.
If you have further questions, don´t hesitate to contact me.
Good luck, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I
} I'm having a stabiliy problem with LR White sections. They curl, expand, } split, etc. during examination on the TEM. My operating conditions are } 60KV, Magnifcations in range of 10 to 45K, trying to keep beam intensity } low. In the past I've tried vac. evap. a layer of carbon which helped some. } I was also thinking of coating the grids with another layer of formvar to } "sandwich" the sections. I would appreciate anyone's thoughts and what they } have tried in solving the stability problem. thanks. } } Tom Bargar } EM Lab, UNMC } Omaha, NE } (402)559-7347
Have you tried making carbon/metal grids (Collodion or Forvar removed). I'm wondering if the metal might help dissipate the heat. Additionally, can you increase your HT (even though the potential energy is higher) which will decrease wavelength which I have found helps with beam sensitive materials. Although, maybe you require low KV for contrast.
I have been using Pella's precut wafers for making cross section samples. I only use them as blanks to build up the cross section. They come with an adhesive sheet that binds them together. When I get further into the middle of the wafer, the edges of the piece have a tendency to chip because of the adhesive. Does anyone know how to release them without this occurring. I am concerned about solvents contaminating them and preventing adhesion.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Dale Shumaker [mailto:dshumake-at-mail.jhmi.edu] Sent: Friday, February 23, 2001 12:49 AM To: JIM ROMANOW Cc: microscopy-at-sparc5.microscopy.com Subject: Re: Si wafers wanted
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Depending on what you call cheap, Ted Pella sells 4" wafers precut to either 5x7 or 5x5mm chips for $61.00 US.
Dale
On Thu, 22 Feb 2001, JIM ROMANOW wrote:
} --------------------------------------------------------------- --------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html }
--------------------------------------------------------------- --------. } } } Hello, } Can someone please give me a source for free or inexpensive Si wafer } rejects? I break them into small squares to use as SEM substrates for nano } particles. } Thank you, } Jim } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } Unit-2131 } Storrs, CT 06269-2131 } bsgphy3-at-uconnvm.uconn.edu } 860 486-2914 voice } 860 486-1936 fax } } }
Dale Shumaker G9 WBSB 725 N Wolfe Street Baltimore, MD 21205
Wilson Lab; Johns Hopkins University School of Medicine, Cell Biology and Anatomy 410-614-2654 410-955-4129 (fax)
we appreciate very much all responses about our request related to a negative analysis of asbestos in building materials. We still might ask some of you "off line" for details or further comments. Thanks a lot!
Silvia Montoro Centro Regional de Investigaciones y Desarrollo de Santa Fe Santa fe Argentina
Try: Roels, F, et al: J. Histochem. Cytochem. 22:442-4,1974 Roels, F, et al: J. Histochem. Cytochem. 27: 1471-7, 1979 Roels, F, et al: Am. J. Med. Genet. 25:257-71, 1986
One of these might have the recipe, however, I have used a procedure published by De Craemer, et al in the Journal of Histochemistry and Cytochemistry, 17:675-680, 1969. They use Tetra-HCl buffer which is more commonly called Tris-HCl buffer. I labeled catalase in liver peroxisomes. I immersion fixed 2mm thick sections of liver 6 hours in 3% Glut made up in 0.1M Cacodylate buffer, then vibratomed 50 um tissue slabs. I used 5-20mg DAB(10mg is good) diluted in 0.1M Tris-HCl buffered to 9.0 pH, add 0.02% H2O2, then incubate 30-60 (you can extend to 120) min at 37C. Rinse in Tris buffer and post-fix in glut, osmicate, dehydrate and embedd in epoxy resin. This was in normal rat liver and the label was good. Make sure you are using really fresh H2O2. Some of the older published articles have EM pictures with very dark label, but these days you should expect less DAB intensity. I have talked to a biochemistry/liver researcher about the decrease in intensity of DAB label, and she was certain that the changes were do to modern manufactured food supplements which are different from what was fed to rats back in the 60's and 70's. One last thing about liver peroxisomes, they are more numerous in the centrolobular areas of the liver.
Good Luck!
} ---------- } From: Tom Januszewski[SMTP:tom.januszewski-at-email.swmed.edu] } Reply To: tom.januszewski-at-email.swmed.edu } Sent: Thursday, February 22, 2001 1:39 PM } To: microscopy-at-msa.microscopy.com } Subject: Teorell-StenhagenBuffer } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Listservers, } } I'm trying to track down a } formulation for Teorell and } Stenhagen buffer. A 2.5mM } concentration of diaminobenzidine } is used with this buffer and } hydrogen peroxide to selectively } stain the catalase in peroxisomes. } The references that I have found } all seem to use this buffer and } reference an article in a German } journal dating to 1938. } Is anyone out there familiar with } this buffer? If so, can you supp;y } the recipe? Is there another buffer } that I can use to effectively stain peroxisomes? } As always, thanks in advance for } your replies. } } Tom Januszewski } Senior Electron Microscopist } UT Southwestern Medical Center at Dallas } Dallas, TX 75390-9039 } 214-648-7291 } FAX: 214-648-6408 } Email: tom.januszewski-at-UTSouthwestern.edu }
I have to correct a mistake, I realized as soon as I sent the e-mail response.......I'm sorry, I have a really bad cold and I mistakenly said there was a change in DAB labeling intensity, which is not what I wanted to say!!
The change is in the number of peroxisomes present in the rat liver due to the dietary changes from the 60's to present day.
} ---------- } From: Tom Januszewski[SMTP:tom.januszewski-at-email.swmed.edu] } Reply To: tom.januszewski-at-email.swmed.edu } Sent: Thursday, February 22, 2001 1:39 PM } To: microscopy-at-msa.microscopy.com } Subject: Teorell-StenhagenBuffer } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Listservers, } } I'm trying to track down a } formulation for Teorell and } Stenhagen buffer. A 2.5mM } concentration of diaminobenzidine } is used with this buffer and } hydrogen peroxide to selectively } stain the catalase in peroxisomes. } The references that I have found } all seem to use this buffer and } reference an article in a German } journal dating to 1938. } Is anyone out there familiar with } this buffer? If so, can you supp;y } the recipe? Is there another buffer } that I can use to effectively stain peroxisomes? } As always, thanks in advance for } your replies. } } Tom Januszewski } Senior Electron Microscopist } UT Southwestern Medical Center at Dallas } Dallas, TX 75390-9039 } 214-648-7291 } FAX: 214-648-6408 } Email: tom.januszewski-at-UTSouthwestern.edu }
We have a question regarding a trouble shooting of chatter/compression on brain sections.
The vibratome we used is VIBRATOME 3000, automated with refrigeration, the mouse brain tissue was fixed in 4% paraformaldehyde, the thickness of section cut was 30-50 um. We have being adjusting either speed or amplitude, however, it doesn't seem to solve the problem.
We would appreciate anyone who would kindly help us to solve this problem.
Many thinks.
Xinran
*********************************************************** Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4. 214A Dallas, TX 75390-9111
I would like to know if there are training courses anywhere in North America on the statistics of quantitative microscopy or stereology. If so, where, or where can I search? Pmiller-at-mbt.com
Dear Bruce, I have not done this etch myself, but my copy of Van der Voort has an etch specifically for SEM of stainless steels: "5 ml Acetic acid 5 ml HNO3 15 ml HCl Aqua regia plus acetic acid. For ferritic grades (of stainless steel). Swab sample 15 s. Deep etch for SEM after 45 s." I hope this helps. You wrote: Question: What solution and method would one use to deep-etch Type 316 STAINLESS STEEL to reveal the inclusions and/or precipitates present in relief. This will be used for SEM analysis. Thankyou in advance for your assistance. Bruce
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
In a message dated 2/23/01 1:26:35 PM, pmiller-at-mbt.com-at-sparc5.microscopy.com writes:
} I would like to know if there are training courses anywhere in North } America on the statistics of quantitative microscopy or stereology. If } so, where, or where can I search? Pmiller-at-mbt.com
The N. C. State Univ. course on quantitative image processing and stereology has been taught for 20 years now. The next session is May 9-11 and there are still some openings available. You can get full information at http://members.aol.com/IPCourse including syllabus, lab materials and books, on-line registration, and a downloadable brochure, or call Cindy Allen at 919 515 8171
I am in the wonderful position of purchasing a new inverted metallograph with digital image acquisition for our laboratory.
I have pretty much decided on the Nikon Epiphot 200 metallograph with brightfield and polarized light modules. Also chosen were the Plan Achromat objectives. Any comments on this choice?
Now the real question. Which digital camera would you buy? I know these things have been discussed greatly here in the past and I have followed them with great interest.
One of our primary wants is live image preview(} 15 fps).
We are leaning towards the Spot RT, although the Insight looks to be equally promising.
Any comments for or against either of these would be appreciated. Also, if anyone has another preference I would appreciate hearing your comments.
Thanks
William T. Giles Sr. Electron Microscopist Met. Lab. Coordinator Henderson Technical Laboratory TIMET PO Box 2128 Henderson NV 89009 Ph: (702)566-4436 Fax: (702)564-9038 E-mail: Bill.Giles-at-timet.com
I know this isn't a microscopy question but suspect many of the microscopy core types on the listserver are responsible for the large, poster-sized format printers. I plan to buy one and would appreciate recommendations (both good and bad ones!). If you have any comments on the workload to maintain and use them, that would also be useful. TIA, tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Hi, I am wondering if anyone knows of a staining method that might allow one to distiguish between a region of proton implanted GaAs and an un-implanted region in a TEM?
Also, does anyone have a URL for a microscopy web forum?
-- David Mathes Department of Materials Science University of Virginia 116 Engineer's Way Charlottesville, VA 22904 804 982 5683
Just a few quick comments on the polymerisation of LR White:
I've been using microwave oven curing of LR White for over two years now, I and others who use this resin for gold labeling studies. The blocks section VERY easily, silver after silver section just roll off the ol' diamond knife edge.
Like others have reported, we also coat grids (300# nickel in these cases, sonicated before use to clean in 10% acetone, 10% HCl, 80% DW, plus two short rinses in acetone) with Formvar, then coat with evaporated carbon. THEN use them to pick up sections. They are very stable under the beam and we have expeienced virtually no stability problems.
Can't really say if microwave curing aids the stability of the sections or not. We do that primarily because it saves time, and the resulting blocks section easily.
We use the Formvar support film primarily to increase stability of sections because of the many gold labeling solutions and rinses the sections on grids have to go through, the carbon coating is primarily for stability under the TEM beam.
For deep etching of austenitic stainless steels use 10% bromine in methanol. see E.M. Mahla and N.A. Nielsen, Carbide Precipitation in Type 304 Stainless Steel-An Electron Microscope Study, Trans. ASM 43:290-322 (1951).
We just got an Epson 9500 large format printer (44 inch). Works great and was highly rated.
Best,
Angela
At 12:14 PM 02/23/2001 -0600, Tom Phillips wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
Has anyone used InSpeck calibration beads from Molecular Probes? I recently purchased InSpeck green and I tried several combinations of microscope and CCD camera settings but I can only read intensity values from 3 concentrations, i.e. if I adjust for the highest concentration I can only image (get readings)3 concentrations and the same is true if I start at the lowest concentration. Also the values I obtained for the 3 concentrations is not linear. Does anyone know if there are other beads available or how else can one calibrate the fluorescence intensity in a fluorescence microscope equipped with a 100W HBO lamp? I would appreciate comments or suggestions.
Thanks,
Cora Bucana Corazon D. Bucana, Ph.D. Dept. Cancer Biology UT M.D.Anderson Cancer Center 1515 Holcombe Blvd, Box 173 Houston, Texas 77030 Phone: 713-792-8106 FAX 713-792-8747
To observe inclusions it is better do not etch steel at all, because etching could hide them (especially deep etch). It is true for both polished cross sections and fracture surfaces. EDS and/or BSE detector could be helpful in the beginning when you are learning how to distinguish inclusions from other features of your sample.
Vladimir
} Question: What solution and method would one use to deep-etch Type 316 } STAINLESS STEEL to reveal the inclusions and/or precipitates } present in } relief. This will be used for SEM analysis. } Thankyou in advance for your assistance. } Bruce } } -------------------------------------------------------------- } ------------- } } }
Tom, I don't know your level of expertise but I'll give you a protocol that I've used for those antibodies on pancreatic cells grwon on coverslips. the protocol is pretty standard with LRW. I use 0.05M tris saline with 1%BSA + 0.05% Tween 20 for all solutions including washes. Place grids section side down on 30 ul drops. Filter all solutions. Note: I use different variations of this protocol but this one is easy to trouble shoot.
1. 15-20 min incubation in 5 to 10 %normal serum of animal used for the secondary antibody. 2. 1- 2 hrs in primary antibody. Try various dilutions, i.e, 1:50 to 1:500. This depends on a number of factors such as background, avidity of the antibody, etc. 1:50 is always a good one to include for one hour. It gives you a feel what changes you may need the next time. In cases where background is too high go overnight with higher dilutions in frig -at- 4C. 3. wash by placing grids on several drops of buffer, i.e. 6 drops 4. 30 -60min in 1:40, 5 or 10nm gold conjugated anti-IgG of the animal that the primary was produced in. 5. wash grids first with 5 drops of buffer followed by 5 drops of distilled water. 6. If you are going to dual label repeat the steps above but now your primary is from a different animal so your first step or blocking step is then different. Also you will used a different size gold particle conjugated secondary. 7. Counter stain with UA and lead. Contrary to what has been said I prefer staining very briefly in UA as a saturated soln in 50% ETOH followed by washing in a descending series of ETOH (50%,25% 10%) to water. I prefer the appearance of the chromatin stained with alcoholic UA. The only problems that I have is over staining if I go much beyond 5 to 10secs. In your case I would try it if you don't feel confident using alcoholic UA used 1% aqueous but stain longer like a minute The staining depends on your preferences. The only problem if you go to long you can't see the gold. Stain with 30 to 60 secs. I have been doing immunolabelling with LRW in everything from seeds to cells to mammary gland without too many problems. You have to be careful at all steps of the way but it works fine. I also stay away from filmed grids if I can and use 300 to 400 mesh nickel grids for many reasons especially when I know structures containing my epitope are common like what you are looking for and the descreased viewable area due to the grids bars is not a factor. LRW in our hands survives well the numerous staining steps. If you have any more questions email me. Good luck.
Hank Adams
Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, TX 77030
-----Original Message----- } From: Tom W Bargar [SMTP:tbargar-at-unmc.edu] Sent: Friday, February 23, 2001 6:17 PM To: Microscopy-at-sparc5.microscopy.com
Anyone have protocols for immunogold labelling of insulin and glucagon in pancreatic islet cells?
Also, many thanks to everyone who responded about the LR White stability problem.
Hallo All! I've noticed a bit of electronic noise starting to creep into our pulse processing electronics, and to eliminate it have turned the gain down a little accordingly. I have some ideas of where it may be coming from but thought I pick the brains of the wealth of experience out there. So, guys n' galls what gives rise to electronic noise in pulse processing electronics. Of course, I could just look up such elimentary stuff on some web page or other, but where's the fun in that? Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
Available with the Center for High Resolution Electron Microscopy at Arizona State University. Position is sponsored by major chemical company and primary focus of the research will be the development and application of environmental electron microscopy to industrially relevant catalyst systems. Areas of particular interest include the study of phase transformations under reaction environments, in situ polymerization, mobility and dynamic microstructural changes. Candidate will have a Ph.D. in material science, material physics, solid-state chemistry or chemical engineering, with extensive experience in catalyst characterization by transmission electron microscopy. Experience in the areas of catalyst synthesis, testing and characterization is preferred. Please submit your resume together and the names of 3 referees to: Dr. Peter A. Crozier, Industrial Associates Program, Center for Solid State Science, Arizona State University, Tempe, AZ 85287-1704, Fax (480) 965-9004, email: crozier-at-asu.edu.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
This thread and a conversation with a colleague here in Dallas recently has gotten me wondering. Has anyone else come across a situation where a formulation for a polymer works fine in one area of the country, but you can't get it to work in another? For instance, years ago when I was working in Minnesota, we used a soft formula for Spurs with fairly good results yet here in Dallas, Tx., I have to use the hardest formula to get a block that is hard enough to section and still some- times the blocks are soft. I also remember that we always had a whole different set of problems to deal with in Minnesota depending on whether it was in the middle of the very dry winter or in the humid spring and summer.
My colleague here in Dallas is trying to use a two part polymer for casting. He has used it successfully some years ago in another part of the country and now he is getting too soft results. I don't know about his protocol, but I have tried adding drierite to the chamber that my blocks are polymerized in, but it has met with limited success.
I also remember a story my old boss told me about his bringing some already cured "epon" blocks that had already been successfully sectioned in Minnesota with him on sabbatical. The blocks were like goo when he tried to section them the second time in Sweden. (This was in the late '60s, early 70's.)
Could the success that you see, Gib, and the lack of success the others are having be related to your location and the "elements"? I have never used a microwave for curing, so maybe that's the difference. Does that shorten the cure time significantly?
Anybody care to comment?
Karen Pawlowski
Gib Ahlstrand wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Microlisters, } } Just a few quick comments on the polymerisation of LR White: } } I've been using microwave oven curing of LR White for over two years now, I } and others who use this resin for gold labeling studies. The blocks section } VERY easily, silver after silver section just roll off the ol' diamond knife } edge. } } Like others have reported, we also coat grids (300# nickel in these cases, } sonicated before use to clean in 10% acetone, 10% HCl, 80% DW, plus two } short rinses in acetone) with Formvar, then coat with evaporated carbon. } THEN use them to pick up sections. They are very stable under the beam and } we have expeienced virtually no stability problems. } } Can't really say if microwave curing aids the stability of the sections or } not. We do that primarily because it saves time, and the resulting blocks } section easily. } } We use the Formvar support film primarily to increase stability of sections } because of the many gold labeling solutions and rinses the sections on grids } have to go through, the carbon coating is primarily for stability under the } TEM beam. } } Gib Ahlstrand
I was analyzing some foam samples and observed small disks (about 25 microns) on the cell walls on one sample. Does anyone out there know what they are? The foam is non-ridged.
I'm Okugawa from Nikon Japan. The adapter is available in China. Could you contact the following woman who is representative in Beijing office.
Product: MXA29005 Contact: Ms. Wang, phone: 010-6515-5637 or 5639
Hisashi Okugawa 1st design department Instrument company in Nikon Corporation Phone: 81-45-853-8568 Fax: 81-45-853-8475 e-mail: okugawa.h-at-nikon.co.jp ----- Original Message ----- } From: "cenpok" {dyy-at-cenpok.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Saturday, February 24, 2001 11:38 AM
Dear Dr.Dong,
This is Okugawa again.
What application do you like to use NIR microscope for? How long wavelength are you interested? Objective effects the image performance. Most of our standard objectives transmit and make good images until 1100nm. Many of them transmits more than 50% and some transmit more than 70%. I may help you technically with your requests. Please contact the e-mail below personally.
Hisashi Okugawa 1st design department Instrument company in Nikon Corporation Phone: 81-45-853-8568 Fax: 81-45-853-8475 e-mail: okugawa.h-at-nikon.co.jp ----- Original Message ----- } From: "cenpok" {dyy-at-cenpok.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Saturday, February 24, 2001 1:17 PM
How would you like to take all of your credit cards, reduce or eliminate the interest, pay 70% less per month, and pay them off 70% sooner?
In addition to the polarized light module, I would suggest getting the DIC (Nomarski) module. This will provide contrast to low contrast etched microstructures such as martensite and acicular structures. We find it very useful for low carbon steels.
Sam Purdy Technical Center National Steel Corp. Trenton, MI spurdy-at-nationalsteel.com
The Department of Materials, University of Oxford, UK, has three vacancies in its Electron Microscope Facility - one senior Engineer and two technicians. The details can be found at http://www.materials.ox.ac.uk/vacancies/vacancieshome.html#SupportStaff.
Closing date for applications is 23rd March 2001.
Please bring this to the attention of anyone who may be interested.
Thanks Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Dear Gib and All, We get calls from all over the country and the increase in technical calls with problems is different for some areas. The time of year and weather conditions can and do play a part in how somethings react and polymerize. It is difficult to tell when or where this will happen and often we recommend drying with dissicant at one point and adding moisture for others. This sounds a little strange and as if the question is being avoided however, you bring up a good point and the changes in the environment should be considered when getting a protocol from a colleague in another area or country. It has caused people to become very angry when they think something is left out or not discussed. in actuality the problem can be the change in environment or even pH of the distilled, DI, or tap water. Histology has these problems often with season changes and the addition of heat or cooling of the laboratory air. Pam Marcum
-----Original Message----- } From: Karen Pawlowski [mailto:Karen.Pawlowski-at-worldnet.att.net] Sent: Saturday, February 24, 2001 1:18 PM To: Gib Ahlstrand Cc: Microscopy-at-sparc5.microscopy.com
Hi Gib and all,
This thread and a conversation with a colleague here in Dallas recently has gotten me wondering. Has anyone else come across a situation where a formulation for a polymer works fine in one area of the country, but you can't get it to work in another? For instance, years ago when I was working in Minnesota, we used a soft formula for Spurs with fairly good results yet here in Dallas, Tx., I have to use the hardest formula to get a block that is hard enough to section and still some- times the blocks are soft. I also remember that we always had a whole different set of problems to deal with in Minnesota depending on whether it was in the middle of the very dry winter or in the humid spring and summer.
My colleague here in Dallas is trying to use a two part polymer for casting. He has used it successfully some years ago in another part of the country and now he is getting too soft results. I don't know about his protocol, but I have tried adding drierite to the chamber that my blocks are polymerized in, but it has met with limited success.
I also remember a story my old boss told me about his bringing some already cured "epon" blocks that had already been successfully sectioned in Minnesota with him on sabbatical. The blocks were like goo when he tried to section them the second time in Sweden. (This was in the late '60s, early 70's.)
Could the success that you see, Gib, and the lack of success the others are having be related to your location and the "elements"? I have never used a microwave for curing, so maybe that's the difference. Does that shorten the cure time significantly?
Anybody care to comment?
Karen Pawlowski
Gib Ahlstrand wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Microlisters, } } Just a few quick comments on the polymerisation of LR White: } } I've been using microwave oven curing of LR White for over two years now, I } and others who use this resin for gold labeling studies. The blocks section } VERY easily, silver after silver section just roll off the ol' diamond knife } edge. } } Like others have reported, we also coat grids (300# nickel in these cases, } sonicated before use to clean in 10% acetone, 10% HCl, 80% DW, plus two } short rinses in acetone) with Formvar, then coat with evaporated carbon. } THEN use them to pick up sections. They are very stable under the beam and } we have expeienced virtually no stability problems. } } Can't really say if microwave curing aids the stability of the sections or } not. We do that primarily because it saves time, and the resulting blocks } section easily. } } We use the Formvar support film primarily to increase stability of sections } because of the many gold labeling solutions and rinses the sections on grids } have to go through, the carbon coating is primarily for stability under the } TEM beam. } } Gib Ahlstrand
I read the literature available on the indicated web site and have a couple of questions.
There seems to be some conflicting information. Under specifications: Image size 3840 x 3072 CCD 1.34 M pixels Document text: 12 million pixels (3046 x 3072) ???????
Is my math wrong or am I missing some pertinent information?
How do you obtain RGB? What does the color array on the chip look like? Or does the camera use filters?
What's the s/n ratio?
Pixel size?
Are there twain drivers available or a Photoshop plugin?
Is any annotaion, image enhancement and/or image analysis software included?
BTW, the Spot RT is $7k also, not twice that price.
William T. Giles Sr. Electron Microscopist Met. Lab. Coordinator Henderson Technical Laboratory TIMET PO Box 2128 Henderson NV 89009 Ph: (702)566-4436 Fax: (702)564-9038 E-mail: Bill.Giles-at-timet.com
} -----Original Message----- } From: Lawrence Kordon [SMTP:nikon-at-jagunet.com] } Sent: Friday, February 23, 2001 1:38 PM } To: Giles, Bill } Cc: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Re: Metallograph and Digital Camera for Light Microscopy } } William, } } Excellent choice on the Nikon Epiphot 200! However, why are you not } considering } the Nikon DXM1200 Digital Camera? It is a whopping 12 Million pixels with } a live } Video Output to a PC and cost under $7000. That is about the half the } price of } the Spot-RT. Also, why would you want to have a cooled CCD anyway? } Metallurgy; } even with Fluorescence and high Mag DIC (Pol) does not require it. Not to } mention this camera can do extremely low light anyway! This Camera is } literally } the hottest product in Microscopy today! We have sold hundreds already } "sight/unseen" and are first shipping them. All the major Image Analysis } companies have or are writing Drivers and it is so popular we have put it } on } other brands microscopes as well! By the way, the US MINT and some NIST } Labs } here is Washington, DC have it. Please check out the specs at.... } http://www.nikonusa.com/usa_product/product.jsp?cat=5&grp=26&productNr=DXM } 1200 } and call you local Nikon Dealer for a Demonstration and Quotation. } } Good Luck, } } Lawrence Kordon } Nikon Instruments, Inc. } Columbia, MD } nikon-at-jagunet.com } } } } "Giles, Bill" wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Good Day Listers, } } } } I am in the wonderful position of purchasing a new inverted metallograph } } with digital image acquisition for our laboratory. } } } } I have pretty much decided on the Nikon Epiphot 200 metallograph with } } brightfield and polarized light modules. Also chosen were the Plan } Achromat } } objectives. Any comments on this choice? } } } } Now the real question. Which digital camera would you buy? I know these } } things have been discussed greatly here in the past and I have followed } them } } with great interest. } } } } One of our primary wants is live image preview(} 15 fps). } } } } We are leaning towards the Spot RT, although the Insight looks to be } equally } } promising. } } } } Any comments for or against either of these would be appreciated. Also, } if } } anyone has another preference I would appreciate hearing your comments. } } } } Thanks } } } } William T. Giles } } Sr. Electron Microscopist } } Met. Lab. Coordinator } } Henderson Technical Laboratory } } TIMET } } PO Box 2128 Henderson NV 89009 } } Ph: (702)566-4436 } } Fax: (702)564-9038 } } E-mail: Bill.Giles-at-timet.com
I've been asked to distribute the following announcement of an available position for a TEM technician at the University of Utah. Anyone who's interested should contact Dr. Kendal Broadie.
Ed King ____________________________
Electron Microscopy Lab Technician
The ideal candidate will have a bachelor degree in a biological science and experience in electron microscopy. Exposusure to neuroscience and/or genetics would be a benefit. The job includes preparing specimens for microscopy, operation of electron microscopes, producing photographic and digital micrographs and analyzing ultrastructural data. Training will be provided. If interested, please submit an application at the University of Utah Human Resources. If you would like more information about job specifics, please contact me (Kendal Broadie) at e-mail: broadie-at-biology.utah.edu
Dr. Kendal S. Broadie (801) 585-9426 (office; 448 Skaggs) Department of Biology (801) 585-9425 (main lab; 450 Skaggs) University of Utah (801) 585-1301 (lab II; 460 Skaggs) 257 South 1400 East (801) 581-4668 (fax) Salt Lake City, Utah 84112-0840 broadie-at-biology.utah.edu
I am looking for micro-crystalline quartz crystals, up to 100 microns in size. Does anybody know of a source? Thank you, Peggy. --
Dear Peggy, I have seen some small quarz crystals while examining river sediment. They're about 1 micron in size, EDS shows lots of Si, and no Na, Al, K, Ca, Fe or Ti--the most common other elements in silicates. I'm also not sure how best to separate them from the other material, although I would think that floating the sediment on an appropriate liquid would work. The Handbook of Chemistry and Physics has a table of Heavy Liquids for Mineral Separation on page E-376 (62nd Edition). Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Two newly funded positions are available to study intracellular protein trafficking using LM and EM immunocytochemistry. Ideally, these positions will be filled by either two postdoctoral fellows or one postdoc and one senior research technician. The projects use similar approaches but are funded by separate sources.
The first project involves using confocal microscopy and electron microscopy, to visualize lipoproteins, lipid synthesizing enzymes, and components of the ubiquitin-proteasome system in hepatic and intestinal cells. Wild-type and heterologous cells will be used to map the location of these proteins in cell compartments and the secretory pathway. Live cell recording using transgenic cells with GFP-labeled constructs are part of this study.
The second project examines the intracellular trafficking and storage of transgenic proteins in maize and other plant seed tissues. The ideal candidate would have experience in plant microscopy and/or light and electron microscopic immunocytochemistry.
Individuals with experience in electron microscopy and prior training in ultramicrotomy are especially encouraged to apply for either position.
Please send a curriculum vitae, cover letter and the names of three references to Dr. Tom Phillips, Division of Biological Sciences, 3 Tucker Hall, University of Missouri, Columbia, MO 65211 7400. Enquires can also be directed to PhillipsT-at-missouri.edu.
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
For your Nikon adapter I would suggest you contact McCrone Microscopes & Accessories from Westmont, IL 60559. They can be contacted on the web at www.mccrone.com or e-mail address info-at-mccrone.com or phone 630-887-7100 or Fax 630-887-7764. If they don't have it they should be able to direct you to who can.
As far as the 1992 Microscopy Analysis of Feedstuffs you can contact the American Oilseed Chemist Society at www.aocs.org. Please contact them and they will be happy to help.
George Falb AOCS/Feed Microscopy Division Chair
----- Original Message ----- } From: "cenpok" {dyy-at-cenpok.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, February 23, 2001 9:38 PM
dear all, does anyone know where the microscope adapter for nikon coolpix 990 could be available?
does anyone know where can I get mannual of microscopial analysis of feedstuffs, 1992 3rd edition, the americian association of feed microscopists?
I'm doing my master in materials science and recently a BSE detector was
installed in the SEM. I'm working with it and I want to know how the detector physically works. The manual just said that the detector is made of semiconducor material. The detector is a GW Electronics system 47 solid state detector. Could some body help me with that description ??
Xinran, I have not had too much experience in vibratoming mouse brains, but I do vibratome rat brains on a regular basis. I have only had problems when the tissue has not been fixed long enough, or when I am cutting 25-30 um sections. Usually turning down the amplitude to a very low setting has worked. Sorry that I couldn't be much more help. Contact me if you have any other specific questions. KElly Randall Henry Ford Hospital kellyr-at-neuro.hfh.edu
On Fri, 23 Feb 2001, Xinran Liu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, colleagues, } } We have a question regarding a trouble shooting of chatter/compression on } brain sections. } } The vibratome we used is VIBRATOME 3000, automated with refrigeration, the } mouse brain tissue was fixed in 4% paraformaldehyde, the thickness of } section cut was 30-50 um. We have being adjusting either speed or amplitude, } however, it doesn't seem to solve the problem. } } We would appreciate anyone who would kindly help us to solve this problem. } } Many thinks. } } Xinran } } *********************************************************** } Xinran Liu, M.D., Ph.D. } Center for Basic Neuroscience } UT Southwestern Medical Center } 6000 Harry Hines Blvd., NA4. 214A } Dallas, TX 75390-9111 } } Phone: (214) 648-1830 } Fax: (214) 648-1801 } E-mail: Xinran.Liu-at-UTsouthwestern.edu } } }
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Pathology lab. listers: are there any new/and/or superior techniques for processing previously formalin fixed tissue for TEM? Thanks, Peggy Harger-Allen |TAB|email:harger-allen-at-indianapolis.va.gov
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id PAA03591 for dist-Microscopy; Mon, 26 Feb 2001 15:55:23 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id PAA03588 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 26 Feb 2001 15:54:53 -0600 (CST) Received: from smtp.rcsis.com (smtp.rcsis.com [208.45.228.18]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id PAA03581 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 26 Feb 2001 15:54:41 -0600 (CST) Received: from ggaugler.gaugler.com (241.dsl108148.rcsis.com [63.108.148.241]) by smtp.rcsis.com (8.9.3/8.9.3) with ESMTP id NAA13323; Mon, 26 Feb 2001 13:49:30 -0800 Message-Id: {5.0.2.1.2.20010226132955.00a8f900-at-pop.calweb.com} X-Sender: gaugler-at-pop.calweb.com (Unverified) X-Mailer: QUALCOMM Windows Eudora Version 5.0.2
The Nikon MX1200 is native 1.2M pixels. It, like current generation digital microscope cameras, do a "mechanical" interpolation to increase final pixel count. One way is to physically move the CCD imager. Each movement results in new image data for that position. This is a bad way to gather more pixels, however. The other way is to keep the CCD fixed but scan the image across it multiple times at different angles.
Zeiss has tried to do mechanical shifting and not done all that good of a job. Looks like Nikon is trying it too. They are up against a basic patent by Pixera. Pixera uses what they call a "Diractor" to scan the image across a fixed CCD.
The Pixera is available as a 1.5M pixel unit or as a 5.8M pixel Diractor unit. Each are also available with Peltier cooling. Like the Nikon, the Pixera uses a dedicated PCI card for PC or Mac. This card, camera and software will do live focus at 12-15 fps on a 733MHz system. It does manual and automatic exposure and also provides a focus indication bar. Also provided is automatic or manual white balance, selectable black balance and multi-size spot exposure metering.
The best way to find out how well a camera performs is to take a high resolution shot at different lighting extremes and do an FFT on the images. Artifacts and garbage will show up if the system produces high pixel count at the expense of image quality.
I use a Pixera Penguin 600CL and like it very much. It is easy to take single shots either as one exposure or averaged or integrated up to 256 exposures. Averaging and Peltier cooling greatly reduces ultimate noise in the image. For simple documentation purposes, this is not necessary. But for images which will undergo further processing, noise and artifacts must be reduced or eliminated. The Pixera has selectable ISO from 50 to 400. I run it at ISO 100. No info on what the Nikon does.
The Pixera also supports Twain interfacing.
gary g.
At 08:57 AM 2/26/01, you wrote:
} Lawrence, } } Thanks for your response. } } I read the literature available on the indicated web site and have a couple } of questions. } } There seems to be some conflicting information. } Under specifications: } Image size 3840 x 3072 } CCD 1.34 M pixels } Document text: 12 million pixels (3046 x 3072) ??????? } } Is my math wrong or am I missing some pertinent information? } } How do you obtain RGB? What does the color array on the chip look like? Or } does the camera use filters? } } What's the s/n ratio? } } Pixel size? } } Are there twain drivers available or a Photoshop plugin? } } Is any annotaion, image enhancement and/or image analysis software included? } } BTW, the Spot RT is $7k also, not twice that price. } } William T. Giles } Sr. Electron Microscopist } Met. Lab. Coordinator } Henderson Technical Laboratory } TIMET } PO Box 2128 Henderson NV 89009 } Ph: (702)566-4436 } Fax: (702)564-9038 } E-mail: Bill.Giles-at-timet.com } } } -----Original Message----- } } From: Lawrence Kordon [SMTP:nikon-at-jagunet.com] } } Sent: Friday, February 23, 2001 1:38 PM } } To: Giles, Bill } } Cc: 'Microscopy-at-MSA.Microscopy.Com' } } Subject: Re: Metallograph and Digital Camera for Light Microscopy } } } } William, } } } } Excellent choice on the Nikon Epiphot 200! However, why are you not } } considering } } the Nikon DXM1200 Digital Camera? It is a whopping 12 Million pixels with } } a live } } Video Output to a PC and cost under $7000. That is about the half the } } price of } } the Spot-RT. Also, why would you want to have a cooled CCD anyway? } } Metallurgy; } } even with Fluorescence and high Mag DIC (Pol) does not require it. Not to } } mention this camera can do extremely low light anyway! This Camera is } } literally } } the hottest product in Microscopy today! We have sold hundreds already } } "sight/unseen" and are first shipping them. All the major Image Analysis } } companies have or are writing Drivers and it is so popular we have put it } } on } } other brands microscopes as well! By the way, the US MINT and some NIST } } Labs } } here is Washington, DC have it. Please check out the specs at.... } } http://www.nikonusa.com/usa_product/product.jsp?cat=5&grp=26&productNr=DXM } } 1200 } } and call you local Nikon Dealer for a Demonstration and Quotation. } } } } Good Luck, } } } } Lawrence Kordon } } Nikon Instruments, Inc. } } Columbia, MD } } nikon-at-jagunet.com } } } } } } } } "Giles, Bill" wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good Day Listers, } } } } } } I am in the wonderful position of purchasing a new inverted metallograph } } } with digital image acquisition for our laboratory. } } } } } } I have pretty much decided on the Nikon Epiphot 200 metallograph with } } } brightfield and polarized light modules. Also chosen were the Plan } } Achromat } } } objectives. Any comments on this choice? } } } } } } Now the real question. Which digital camera would you buy? I know these } } } things have been discussed greatly here in the past and I have followed } } them } } } with great interest. } } } } } } One of our primary wants is live image preview(} 15 fps). } } } } } } We are leaning towards the Spot RT, although the Insight looks to be } } equally } } } promising. } } } } } } Any comments for or against either of these would be appreciated. Also, } } if } } } anyone has another preference I would appreciate hearing your comments. } } } } } } Thanks } } } } } } William T. Giles } } } Sr. Electron Microscopist } } } Met. Lab. Coordinator } } } Henderson Technical Laboratory } } } TIMET } } } PO Box 2128 Henderson NV 89009 } } } Ph: (702)566-4436 } } } Fax: (702)564-9038 } } } E-mail: Bill.Giles-at-timet.com
I am going off my recollection, which can sometimes be a dangerous thing, but it should be a start and may be adequate.
I believe your GW BSE detector is very similar in principle to solar voltaic cells. Incident photons create electron-hole pairs which generate a current/voltage. You will find that such detectors are swamped when the SEM chamber light is turned on or the chamber is opened to room light.
In the other camp are Robinson style BSE detectors. These are better considered as scintillator type detectors. Incident electrons generate a pulse of light which is directed down a light pipe to a photo-multiplier tube. This kind can generate a signal suitable for TV-rate imaging, but may not be very sensitive to low signal levels. The former kind may be more sensitive, but are slower responding and suitable primarily for slow scans.
The most basic of SEM books should describe the difference between backscattered and secondary electrons. The intensity of the BSE signal is directly related to the atomic number of the phase under the beam. Therefore, the BSE signal gives indications of compositional variation even without EDX. I also remember seeing some articles that talked of doing spectroscopy on the BSE signal (i.e., measuring energy of the scattered electrons) and relating that to the material doing the scattering. However, basic BSE systems are not setup for such measurements.
Hope this gets you started.
At 11:05 AM 2/26/2001 -0600, you wrote:
} Hello all, } } I'm doing my master in materials science and recently a BSE detector was } } installed in the SEM. I'm working with it and I want to know how the } detector } physically works. The manual just said that the detector is made of } semiconducor material. The detector is a GW Electronics system 47 solid } state detector. Could some body help me with that description ?? } } } Laura Lopez } CCMC-UNAM } Ensenada, B.C. Mexico
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Laura, The BSE detector is basically a solar cell and is able to generate a current from the energy released by backscattered eleectrons impinging on its surface. It is fairly fragile, so don't run a specimen up into it. Also GW has done a lot of work on this and a regular solar cell won't work because it has too much capacitance. These are very thin. I also seem to remember that they went to a cell that was doped opposite to the doping normally found in solar cells (P instead of N or vice versa). The signal is then run through a preamplifier and on to the main amplifier. For TV rates (if that was set up) I believe they reverse bias the cell to further reduce capacitance and increase gain (sensitivity). Secondary electrons don't have nearly enough energy to cause the cell to react.
Ken Converse owner Quality Images Delta, PA
Laura Lopez wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hello all, } } I'm doing my master in materials science and recently a BSE detector was } } installed in the SEM. I'm working with it and I want to know how the } detector } physically works. The manual just said that the detector is made of } semiconducor material. The detector is a GW Electronics system 47 solid } state detector. Could some body help me with that description ?? } } } Laura Lopez } CCMC-UNAM } Ensenada, B.C. Mexico } } } }
There VERY OLD, however they were manufactured by the Cambridge Instrument Company, now part of LEO. (Cambribge SEMs). They may be able to help you. At one stage the Huxley Mk2 were distributed by LKB, now part of the Leica Instruments Company, they also may be able to help you. Regards JVN
Steve Widing wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello List, } } Does anyone have a contact or supplier for parts for a Huxley } Ultra Microtome MK2? } } Thanks, } } Steve Widing } Temple University
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
-----Original Message----- } From: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Monday, February 26, 2001 4:39 PM To: "HARGER-ALLEN.MARGARET_J-at-INDIANAPOLIS.VA.GOV"-at-sparc5.microscopy.com
Pathology lab. listers: are there any new/and/or superior techniques for processing previously formalin fixed tissue for TEM? Thanks, Peggy Harger-Allen |TAB|email:harger-allen-at-indianapolis.va.gov
I was wondering if anyone might have an extra filter cube holder laying around for an older Nikon Diaphot inverted microscope.
Any help would be greatly appreciated.
Thanks,
Raj
********************************************** Dr. Raj Lartius, CEO NovaScan Technologies Iowa State University Research Park 2501 North Loop Drive Ames, Iowa USA 50010
Email: rlartius-at-novascan.com Voice: 515-795-3164 Fax: 515-795-4414 ********************************************** "Innovative Tools to Explore the Microworld"
Email: risports-at-aol.com Name: Keenan Kelly School: Veterans Memorial HS
Question: I recently bought an American Optical Model 150 microscope used on eBay and would like to add phase contrast to it.
Question 1.) Why can't I find a web site or other company information about American Optical on the web?
Question 2.) Are objectives and condensers interchangeable between microscopes from different manufacturers? If not all, then how do you find out which are?
Email: maegan-at-ualr.edu Name: Marilyn Egan School: University of Arkansas at Little Rock
Question: Are you familure with dispersion staining techniques? I need information regarding microscope set up and grain orientation with respect to the annular and central stops.
Fix in glutaraldehyde, dehydrate through a gradient ethanol and acetone series, critical point dry, glue to a stub and sputter-coat or --attach to a specimen mount for cryo-sem, freeze, etch by raising the temperature to -100C, sputter-coat and image.
I am looking for a nice glue to fix the specimen to the tripod polisher for wedge polishing. The "super glue" stuff does not work well. The specimen falls off too easily.
I would appreciate very much if anyone could send me some good idea.
This week, our CPD was broken. I was planning to CPD some samples that day. The samples are now in acetone. My question : how long can these samples stay in acetone ? The samples are epithelial cells from duodenum grown on glass.
Nikon DXM1200 has 1.3M pixels CCD. With 3 x 3 times "Diractor", it makes 12M pixels images. Because of the color array, 2 pixels should be moved totally. DXM1200 moves two times every two thirds pixels. Pixera moves 1 time every one pixel or three times every half pixels. Please visit the following web to get more information
http://www.microscopyu.com/
Hisashi Okugawa 1st design department Instrument company in Nikon Corporation Phone: 81-45-853-8568 Fax: 81-45-853-8475 e-mail: okugawa.h-at-nikon.co.jp ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Giles, Bill" {William.Giles-at-TIMET.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 27, 2001 6:52 AM
Dear Sam,
DXM1200 is high sensitive enough to get good DIC images or some fluorescence images.
Hisashi Okugawa 1st design department Instrument company in Nikon Corporation Phone: 81-45-853-8568 Fax: 81-45-853-8475 e-mail: okugawa.h-at-nikon.co.jp ----- Original Message ----- } From: "Purdy, Sam" {SPurdy-at-nationalsteel.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, February 26, 2001 11:28 PM
If those cells were for TEM sections, I would be very concerned and re-hydrate them back to 70% alcohol immediately. For SEM lipid extractions matters less but you may well get some shrinkage. Its less urgent, but I would take those cells back to 70%. At that and refrigerated you SHOULD see no real change for some weeks. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, February 27, 2001 7:56 PM, Bart De Pauw [SMTP:Bart.DePauw-at-rug.ac.be] wrote: } } Hello Everyone, } } This week, our CPD was broken. I was planning to CPD some samples that } day. The samples are now in acetone. My question : how long can these } samples stay in acetone ? The samples are epithelial cells from duodenum } grown on glass. } } greetings, } } De Pauw Bart } Belgium }
I have been given mouse eyes in 2% paraformaldehyde/2.5% glutaraldehyde buffered with Millonig's phosphate buffer. I was asked to rate the lens for opacity and then to fix the eyes in tact and section them. Any suggestions would be appreciated. Thanks.
{html} {DIV} Hi all, {/DIV} {DIV} I'm from the university of Massachusetts, Lowell. We are looking for a microtome to replace the old one we have. does anyone have a old microtome they want to sell off or just trying to get rid of? {/DIV} {DIV} Praveena {/DIV} {DIV} Department of Chemical Engineering {/DIV} {DIV} Unversity of Massachusetts, Lowell {/DIV} {DIV} Lowell , MA {/DIV} {DIV} PH: 978-934-3411 {/DIV} {DIV} {/DIV} {br clear=all} {hr} Get Your Private, Free E-mail from MSN Hotmail at {a href="http://www.hotmail.com"} http://www.hotmail.com {/a} . {br} {/p} {/html}
Dear rad0, I have looked at fungus and fungus spores on my SEM and they are very simple to prepare. Just stick them to a stub by sticky tab or glue and then gold coat. They are very robust and they don't need any fixing or dehydration. At 06:06 PM 2/26/01 -0600, you wrote: } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks... } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve } } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
There has been a lot of discussion over the past on this subject. The boys from IBM are the experts. (There's your plug, Ron.) Since nobody has answered this yet, I'll give you my summary. Ron, If I say anything wrong, please correct me.
Don't use any type of super glue gel! It doesn't work, you have to use the thin stuff.
The IBM folks (Ron et al.) recommend Ross super glue, but that is sometimes difficult to locate in other parts of the country.
I have had good success with the Alpha superglue from Electron Microscopy Sciences and with Loctite superglue available almost anywhere. Buy it from a place that has a high turnover rate of it so that it is fresh. The IBM guys recommend keeping the stuff unopened in the refrigerator for any length of time. Once it has been opened, you can't use it for very long (a day or two). I have used the Loctite a little longer because it has a very good sealing top.
I think that pressure on the sample while it cures is important, but the IBM guys just wick the extra stuff away from the sample and don't put a lot of pressure on the sample.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Csaba Cserhati [mailto:cserhati-at-delfin.klte.hu] Sent: Tuesday, February 27, 2001 4:22 AM To: Microscopy-at-sparc5.microscopy.com Subject: Tripod Polisher
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I am looking for a nice glue to fix the specimen to the tripod polisher for wedge polishing. The "super glue" stuff does not work well. The specimen falls off too easily.
I would appreciate very much if anyone could send me some good idea.
At 9:37 AM -0500 2/27/01, stacey andringa wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
*********************** Hi Stacey,
If you must leave the eyes intact, I would suggest REALLY prolonging everything: at least 30 minutes in each alcohol, and take about a day & a half for the infiltration. Use a rotator during all steps. I've had better luck using Epon-Araldite for eyes, although I've also used Spurr's.
If its at all possible, find out if you (or the person you are doing this for) can make a few small holes through the sclera to give better access of solutions to the internal structures. eyes are very complex, with lots of layers of different cell types and different consistencies, so you really need to get your reagents to penetrate thoroughly. The vitreous, in the center, is frequently problematic and one of the main reasons for needing the long steps.
Eyes are not easy, but when you've don a good job, they are beautiful. Stain your 1 micron sections with methylene blue...gorgeous!
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
What are some good demonstration specimens that clearly show phase halo and shading off? I am especially interested in demonstrating shading off.
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718 http://www.pathology.med.unc.edu/path/microscopy/welcome.htm
CONFOCAL MICROSCOPIST POSITION OPEN AT GENZYME CORPORATION (FRAMINGHAM, MA)
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I have looked extensively at a wild strain of Aspergillus many years ago. I grew the fungus on sterilized glass slides with half-strength saubourod-dextrose agar for at t least two days and up to 21 days. I cut out pieces of the agar culture and fixed in 1% osmium fumes fix overnight under cover, then a standard dehydration in ETOH, two changes of amyl acetate and then critical point drying. These were sputter coated several times as these were difficult to view without charging. I never published this, but I dragged the paper out recently after 22 or so years to see if I could do SEM on Candida. Funny you should also ask! Good luck and if you have any other question, you can reach me at Bplowman-at-sfuop.edu.
Barbara L. Plowman Univ. of the Pacific School of Dentistry 2155 Webster Rm 632 San Francisco, CA 94115 ph: 415-929-6692 FAX: 415-929-6654 email: Bplowman-at-sfuop.edu
Can anyone tell me if the required currents for the different brands of LaB6 filaments are significantly different?
The reason that I am asking this is that it appears that on my JEOL 2000FX that I can not fully saturate the LaB6 filament when it is set up correctly, i.e. tip to whenelt distance, centering, biasing. There appears to be a limit at which no more current is available as you turn up the filament current. If I lower the bias, I can saturate the filament, but the emission is too low. I seem to recall a discussion some time ago about whether the filament power on some of the older 2000FX had enough juice to do the job, but I may be mistaken.
I am using a new Denka filament. I have a new titanium wehnelt. It is doing the same as a used and a new Denka filament that I had in using the old SS wehnelt assembly. There are no problems when a tungsten filament is used. My thought is that if another brand of filament requires less current to heat (say about the same as the tungsten filament) that I might be able to use that.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Group, At the upcoming M&M Conference in Long Beach (5/9 August), Microscopy Today will again hold a "Just For Fun Micrograph Contest". Prizes will be $300, $200 and $100 respectively for first second and third prizes. If interested in participating, kindly contact me off line and I will provide complete information. See you in Long Beach, Don Grimes, Microscopy Today
I was wanting to know whether anyone has used methyl salicylate to clear fixed slices of plant tissue, and if so, what in their experience is the best method to use.
a colleague would like to simulate HRES images to compare with those recorded using our JEOL 2000fxII TEM. For this he needs to know the objective lens spherical aberration coefficient (Cs). The polepiece we have is the AHP20L. Does anyone know what the Cs value should be for this instrument?
Another parameter he requires is the beam semi-convergence (in mrad). How is this parameter defined and how can we measure it?
I would appreciate any advice you can offer. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists, } } I am looking for a nice glue to fix the specimen to the tripod polisher for } wedge polishing. The "super glue" stuff does not work well. The specimen falls } off too easily. } } I would appreciate very much if anyone could send me some good idea. } } Best Regards } Csaba Cserhati } Can you fix your specimen with epoxy resin?
I like to glue specimens with epoxy resin because the substance does not harden too fast, and you always have time to find the best location for your sample, so that the latter does not break.
I once read a study which claimed 100% was better that the usual 70%. I cannot remember the authors etc.
Dave
On Tue, 27 Feb 2001 21:39:29 +1000 Jim at ProSciTech {jim-at-proscitech.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } If those cells were for TEM sections, I would be very concerned and re-hydrate } them back to 70% alcohol immediately. For SEM lipid extractions matters less } but you may well get some shrinkage. Its less urgent, but I would take those } cells back to 70%. At that and refrigerated you SHOULD see no real change for } some weeks. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Tuesday, February 27, 2001 7:56 PM, Bart De Pauw } [SMTP:Bart.DePauw-at-rug.ac.be] wrote: } } } } Hello Everyone, } } } } This week, our CPD was broken. I was planning to CPD some samples that } } day. The samples are now in acetone. My question : how long can these } } samples stay in acetone ? The samples are epithelial cells from duodenum } } grown on glass. } } } } greetings, } } } } De Pauw Bart } } Belgium } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Mark A long time ago I needed to clear corn roots with attached parasitic roots to make it possible to trace vascular connections between host and parasite. If I remember correctly the process I used employed xylol instead of salicylate:
Decolorize and soften material in 1% hypochlorite bleach or in 0.5M NaOH. Time (hours to days) and choice of bleach or alkali depends on thickness and colour of the material.
After material is sufficiently translucent, dehydrate in ethanol to 100% and clear by immersing in xylol (take all precautions with this). My recollection is that some materials became glassy clear, to such an extent that it was necessary to stain the vascular bundles with a trace of Safranin O during the 100% ethanol step.
Jan
MARK JEFFREY TALBOT wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } I was wanting to know whether anyone has used methyl salicylate to clear } fixed slices of plant tissue, and if so, what in their experience is the } best method to use. } } Thanks in advance, } } Mark Talbot.
-- Prof Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
I am interested in a SEM that automatically can detect and quantify the number and chemical composition of inclusions present on a polished steel surface.
The SEM should manage to scan an area automatically and detect all the inclusions, which fulfill some user defined requirements e.g. a size over a certain limit or a special chemical composition
Does anybody have experience of such system and what manufactures are there ?
The Society of Electron Microscope Technology (SEMT), is holding a one day meeting in London (England) on 28th March 2001 to include the following topics:
Cryo TEM Low Vacuum/Low Voltage Microscopy GFP Stereology Confocal and 2-Photon Microscopy Digital Imaging in Microscopy
All are welcome and further details are shown on the attachment. As a committee member I can supply more information but it might be quicker to go direct to the secretary David McCarthy, e-mail address also on the attachment,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd ------=_NextPart_000_01C0A174.09670A80 Content-Type: application/octet-stream; name="SEMTMarch28" Content-Transfer-Encoding: base64 Content-Disposition: attachment; filename="SEMTMarch28"
Most EDS manufacturers offer this function as a part of their software. Defining the inclusion positions is a key issue. Most often this is done using BSE imaging by choosing a grayscale range (from histogram) of inclusions while excluding the matrix. It may be an option for some. The most simple method uses beam sweep control by the EDS/imaging system. Some "integrated" EDS/SEM systems may also offer stage control. Haven't looked in detail at the various manufacturer's specs for a while, so hesitate to be more detailed. They, I am sure, would be glad to give you the details!
Woody
} ---------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } } I am interested in a SEM that automatically can detect } and quantify the number and chemical composition of } inclusions present on a polished steel surface. } } The SEM should manage to scan an area automatically } and detect all the inclusions, which fulfill some user defined } requirements e.g. a size over a certain limit or a special } chemical composition } } Does anybody have experience of such system and } what manufactures are there ? } } Best regards } Joakim Fagerlund } } } } }
I agree with Scott (below). I've mangled a number of samples, in various ways, learning the tripod technique but have never had a complete (rare partial de-wets can be repaired) glue failure using the Loctite product. It's available in a "pen" type applicator which seems to have the best bench life of any of the applicators I've used. My tube usually is swiped off the bench long before it goes bad. One of the advantages to the cyanoacrylic cement is it dissolves, in a reasonable time, in acetone. If you were to use a crosslinked epoxy, you'll need to devise a cleaning scheme that will exhaustively remove the epoxy without altering your sample. Good luck.
John
John W. Heckman MSM Department 3505 Engineering Building Michigan State University East Lansing, MI 48824-1226 heckman-at-pilot.msu.edu
517-353-9719 (work) 517-353-9842 (dept. fax)
"Walck, Scott D." wrote:
} } There has been a lot of discussion over the past on this subject. The boys from IBM are the experts. (There's your plug, Ron.) Since nobody has answered this yet, I'll give you my summary. Ron, If I say anything wrong, please correct me. } } Don't use any type of super glue gel! It doesn't work, you have to use the thin stuff. } } The IBM folks (Ron et al.) recommend Ross super glue, but that is sometimes difficult to locate in other parts of the country. } } I have had good success with the Alpha superglue from Electron Microscopy Sciences and with Loctite superglue available almost anywhere. Buy it from a place that has a high turnover rate of it so that it is fresh. The IBM guys recommend keeping the stuff unopened in the refrigerator for any length of time. Once it has been opened, you can't use it for very long (a day or two). I have used the Loctite a little longer because it has a very good sealing top. } } I think that pressure on the sample while it cures is important, but the IBM guys just wick the extra stuff away from the sample and don't put a lot of pressure on the sample. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) }
I responded to the requester off line and said that most any super glue works well. I should have added that "super glue" is defined as a "cyanoacrylate," which should be printed on the bottle. We've never tried a "gel' but if Scott doesn't like it that's good enough for us. The main emphasis of my response was the absolute requirement for a clean surface for the glue to bond to. Lack of cleanliness is the biggest cause of specimens "falling off too easily." The bonding surfaces of the tool and the specimen *must* be free of any contamination films. Inspect the surfaces with reflected light. Water from the tap and impure solvents are to blame.
With regard to the glue. We stopped using Ross awhile back. Currently we have Loctite and ND Industries Super Glue in the lab.
Ron
Don't use any type of super glue gel! It doesn't work, you have to use the thin stuff.
The IBM folks (Ron et al.) recommend Ross super glue, but that is sometimes difficult to locate in other parts of the country.
I have had good success with the Alpha superglue from Electron Microscopy Sciences and with Loctite superglue available almost anywhere. Buy it from a place that has a high turnover rate of it so that it is fresh. The IBM guys recommend keeping the stuff unopened in the refrigerator for any length of time. Once it has been opened, you can't use it for very long (a day or two). I have used the Loctite a little longer because it has a very good sealing top.
I think that pressure on the sample while it cures is important, but the IBM guys just wick the extra stuff away from the sample and don't put a lot of pressure on the sample.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Csaba Cserhati [mailto:cserhati-at-delfin.klte.hu] Sent: Tuesday, February 27, 2001 4:22 AM To: Microscopy-at-sparc5.microscopy.com Subject: Tripod Polisher
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
I am looking for a nice glue to fix the specimen to the tripod polisher for wedge polishing. The "super glue" stuff does not work well. The specimen falls off too easily.
I would appreciate very much if anyone could send me some good idea.
I would like to ask some basic things of those cameras that scan the CCD to get more resolution than the specific CCD really offers. I can understand that this can work with color CCDs that have the color mask (array or mosaic or whatever it is called) on the chip and cannot reach the full resolution of the CCD. However, according to my knowledge the "scanning" cannot make the resolution better than with the same chip used as black-and-white camera (without the color mask). So the scanning is done just to compensate against the loss of resolution caused by the color mask. Or has somebody really invented some new rules. Could somebody please advise me, if I have missed something here.
-----Original Message----- From: Hisashi Okugawa [SMTP:okugawa.h-at-nikon.co.jp] {mailto:[SMTP:okugawa.h-at-nikon.co.jp]} Sent: Tuesday, February 27, 2001 12:18 PM To: Giles, Bill; Gary Gaugler Cc: MSA listserver Subject: Re: Metallograph and Digital Camera for Light Microscopy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html}
Nikon DXM1200 has 1.3M pixels CCD. With 3 x 3 times "Diractor", it makes 12M pixels images. Because of the color array, 2 pixels should be moved totally. DXM1200 moves two times every two thirds pixels. Pixera moves 1 time every one pixel or three times every half pixels. Please visit the following web to get more information
Hisashi Okugawa 1st design department Instrument company in Nikon Corporation Phone: 81-45-853-8568 Fax: 81-45-853-8475 e-mail: okugawa.h-at-nikon.co.jp {mailto:okugawa.h-at-nikon.co.jp} ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com {mailto:gary-at-gaugler.com} } To: "Giles, Bill" {William.Giles-at-TIMET.com {mailto:William.Giles-at-TIMET.com} } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com {mailto:Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, February 27, 2001 6:52 AM Subject: RE: Metallograph and Digital Camera for Light Microscopy
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -----------------------------------------------------------------------. } } } The Nikon MX1200 is native 1.2M pixels. It, like current } generation digital microscope cameras, do a "mechanical" } interpolation to increase final pixel count. One way is } to physically move the CCD imager. Each movement } results in new image data for that position. This is a bad } way to gather more pixels, however. The other way is to } keep the CCD fixed but scan the image across it multiple } times at different angles. } } Zeiss has tried to do mechanical shifting and not done } all that good of a job. Looks like Nikon is trying it too. } They are up against a basic patent by Pixera. Pixera } uses what they call a "Diractor" to scan the image } across a fixed CCD. } } http://www.pixera.com/PixeraCatalog/Penguin/Penguin.htm {http://www.pixera.com/PixeraCatalog/Penguin/Penguin.htm} } } The Pixera is available as a 1.5M pixel unit or as a 5.8M pixel } Diractor unit. Each are also available with Peltier cooling. } Like the Nikon, the Pixera uses a dedicated PCI card for } PC or Mac. This card, camera and software will do } live focus at 12-15 fps on a 733MHz system. It does } manual and automatic exposure and also provides a focus } indication bar. Also provided is automatic or manual } white balance, selectable black balance and multi-size } spot exposure metering. } } The best way to find out how well a camera performs is } to take a high resolution shot at different lighting extremes } and do an FFT on the images. Artifacts and garbage will } show up if the system produces high pixel count at the } expense of image quality. } } I use a Pixera Penguin 600CL and like it very much. It } is easy to take single shots either as one exposure or } averaged or integrated up to 256 exposures. Averaging } and Peltier cooling greatly reduces ultimate noise in the } image. For simple documentation purposes, this is } not necessary. But for images which will undergo } further processing, noise and artifacts must be reduced } or eliminated. The Pixera has selectable ISO from 50 to 400. } I run it at ISO 100. No info on what the Nikon does. } } The Pixera also supports Twain interfacing. } } gary g. } } At 08:57 AM 2/26/01, you wrote: } } } Lawrence, } } } } Thanks for your response. } } } } I read the literature available on the indicated web site and have a couple } } of questions. } } } } There seems to be some conflicting information. } } Under specifications: } } Image size 3840 x 3072 } } CCD 1.34 M pixels } } Document text: 12 million pixels (3046 x 3072) ??????? } } } } Is my math wrong or am I missing some pertinent information? } } } } How do you obtain RGB? What does the color array on the chip look like? Or } } does the camera use filters? } } } } What's the s/n ratio? } } } } Pixel size? } } } } Are there twain drivers available or a Photoshop plugin? } } } } Is any annotaion, image enhancement and/or image analysis software included? } } } } BTW, the Spot RT is $7k also, not twice that price. } } } } William T. Giles } } Sr. Electron Microscopist } } Met. Lab. Coordinator } } Henderson Technical Laboratory } } TIMET } } PO Box 2128 Henderson NV 89009 } } Ph: (702)566-4436 } } Fax: (702)564-9038 } } E-mail: Bill.Giles-at-timet.com {mailto:Bill.Giles-at-timet.com} } } } } } -----Original Message----- } } } From: Lawrence Kordon [SMTP:nikon-at-jagunet.com] {mailto:[SMTP:nikon-at-jagunet.com]} } } } Sent: Friday, February 23, 2001 1:38 PM } } } To: Giles, Bill } } } Cc: 'Microscopy-at-MSA.Microscopy.Com' } } } Subject: Re: Metallograph and Digital Camera for Light Microscopy } } } } } } William, } } } } } } Excellent choice on the Nikon Epiphot 200! However, why are you not } } } considering } } } the Nikon DXM1200 Digital Camera? It is a whopping 12 Million pixels with } } } a live } } } Video Output to a PC and cost under $7000. That is about the half the } } } price of } } } the Spot-RT. Also, why would you want to have a cooled CCD anyway? } } } Metallurgy; } } } even with Fluorescence and high Mag DIC (Pol) does not require it. Not to } } } mention this camera can do extremely low light anyway! This Camera is } } } literally } } } the hottest product in Microscopy today! We have sold hundreds already } } } "sight/unseen" and are first shipping them. All the major Image Analysis } } } companies have or are writing Drivers and it is so popular we have put it } } } on } } } other brands microscopes as well! By the way, the US MINT and some NIST } } } Labs } } } here is Washington, DC have it. Please check out the specs at.... } } }
} } } 1200 } } } and call you local Nikon Dealer for a Demonstration and Quotation. } } } } } } Good Luck, } } } } } } Lawrence Kordon } } } Nikon Instruments, Inc. } } } Columbia, MD } } } nikon-at-jagunet.com {mailto:nikon-at-jagunet.com} } } } } } } } } } } } } "Giles, Bill" wrote: } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com}
} } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } } } } -----------------------------------------------------------------------. } } } } } } } } Good Day Listers, } } } } } } } } I am in the wonderful position of purchasing a new inverted metallograph } } } } with digital image acquisition for our laboratory. } } } } } } } } I have pretty much decided on the Nikon Epiphot 200 metallograph with } } } } brightfield and polarized light modules. Also chosen were the Plan } } } Achromat } } } } objectives. Any comments on this choice? } } } } } } } } Now the real question. Which digital camera would you buy? I know these } } } } things have been discussed greatly here in the past and I have followed } } } them } } } } with great interest. } } } } } } } } One of our primary wants is live image preview(} 15 fps). } } } } } } } } We are leaning towards the Spot RT, although the Insight looks to be } } } equally } } } } promising. } } } } } } } } Any comments for or against either of these would be appreciated. Also, } } } if } } } } anyone has another preference I would appreciate hearing your comments. } } } } } } } } Thanks } } } } } } } } William T. Giles } } } } Sr. Electron Microscopist } } } } Met. Lab. Coordinator } } } } Henderson Technical Laboratory } } } } TIMET } } } } PO Box 2128 Henderson NV 89009 } } } } Ph: (702)566-4436 } } } } Fax: (702)564-9038 } } } } E-mail: Bill.Giles-at-timet.com {mailto:Bill.Giles-at-timet.com} } }
An Email attachment got through the filter this AM.
Although it was a microscopy relevant posting it still contained an attachment. This message about a meeting in London managed to find it's way through a loop-hole in the filter software. I believe that I have plugged that hole now. (I guess I should thank you Terry, since your posting allowed me to find the loop-hole).
As a safety issue let me remind you that you should scan all Email attachments for viruses before you open them as it is possible for viruses to be attached to messages even if the author did not intend it.
As a safety issue, we "try" to reject all postings having attachments and succeed most times. Please remember to send all your Email messages as simple ASCII text embedded in the body of your message. Do not embedd HTML in your messages as some Email programs (MS Outlook in particular) converts this to an attachment in many situations, if this happens then as an attachment the HTML embedded message will then get rejected.
In case your curious, of the 2, 234 messages have triggered the Filter and been rejected in the last 2 years. The vast majority of these were true junk mail.
Remember if you post a message and it gets caught and tagged by the filter as suspect mail, READ THE REJECTION MESSAGE and then follow the directions so that I can sort out the problem related to your posting . Most of the rejections are true junk mail, but a few of you will get caught by accident, this is the price we all have to pay due to the junk mailers.
I apologize for this message. I have not been receiving messages from the list since the second week in February and I am trying to establish why. Again my apologies for cluttering your email boxes.
____________________________________________ Massimo Sassaroli Dept. of Physiology and Biophysics, Box 1218 Mount Sinai School of Medicine One Gustave L. Levy Place New York, NY 10029 Tel:(212) 241-9512 FAX:(212) 860-3369 email: massimo-at-inka.mssm.edu
On the convergence semi-angle, you can measure this in the diffraction pattern IF you are working close to crossover (i.e. beam at crossover on the specimen). Then, the diffraction pattern will consist of disks, and the semi-angle for convergence is the radius of the disks, which you can convert to angle in radians by scaling with respect to the other reflections in the pattern (when you index, you get the Bragg angle if the lattice parameters are known). This angle will scale with the condenser aperture radius. (For more detail, see O'Keefe and Sanders, Acta Cryst A31 (1975) p. 307.)
It is likely that on a modern TEM you are not working very close to beam crossover on the sample. You may safely assume that the convergence at crossover is an upper limit. However, when you decondense the beam you are producing a situation where incident angle varies as a function of position on the sample, and locally the convergence may be much smaller than the total convergence over the entire illuminated area. In the back focal plane, each disk now contains an image of the sample, which goes together with the previous statement.
If you refocus the diffraction pattern so that each spot is a sharp filament image you will see that decondensing demagnfies the filament, so it is (locally) decreasing the convergence angle significantly. The convergence angle now scales not with the condenser aperture but with the spot size. To measure it you may refocus the diffraction pattern to make each spot an in-focus filament image, then measure the semi-angle of the filament images.
I hope this is not more confusing here than helpful!
Regards,
Wharton Sinkler UOP LLC Des Plaines, IL
} -----Original Message----- } From: Mark Blackford [SMTP:mgb-at-ansto.gov.au] } Sent: Tuesday, February 27, 2001 9:43 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: JEOL 2000fxII aberration coefficients } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } a colleague would like to simulate HRES images to compare with those } recorded using our JEOL 2000fxII TEM. For this he needs to know the } objective lens spherical aberration coefficient (Cs). The polepiece } we have is the AHP20L. Does anyone know what the Cs value should be } for this instrument? } } Another parameter he requires is the beam semi-convergence (in mrad). } How is this parameter defined and how can we measure it? } } I would appreciate any advice you can offer. Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily } represent the official views of ANSTO from which this message was } conveyed.
I am interested in doing quantitative stereo microscopy using a TEM and am looking for software that may be helpful in such an endeavor. I anticipate the primary application will be examining the spatial arrangement of linear defects. I have not been able to find a software designed directly for quantitative stereo TEM. I have come across one commercial software that can be used for SEM surface topography that may be applicable. It seems like there must be other software packages out there used for applications such as surface mapping, electron tomography, or even astronomy that could be useful. Also, as I am new to stereo microscopy, any words of wisdom would be appreciated.
I would like to ask some basic things of those cameras that scan the CCD to get more resolution than the specific CCD really offers. I can understand that this can work with color CCDs that have the color mask (array or mosaic or whatever it is called) on the chip and cannot reach the full resolution of the CCD. However, according to my knowledge the "scanning" cannot make the resolution better than with the same chip used as black-and-white camera (without the color mask). So the scanning is done just to compensate against the loss of resolution caused by the color mask. Or has somebody really invented some new rules. Could somebody please advise me, if I have missed something here.
Dear Ari, You are, indeed, missing something. I expect that there is a less obscure referrence, but the one that I know of is a paper by Atsushi Imiya in Vol. 101 (1997) of Advances in Imaging and Electron Physics. The paper, Formal Polynomials for Image Processing, goes from p. 99-142, and includes a section from p. 125-135 on subpixel resolution and pyramid transforms. The author's address is Department of Information and Computer Sciences, Faculty of Engineering, Chiba University, Yayoi-cho, Chiba 260, Japan. If you can't find the volume, you can write for a reprint. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Could anyone please recommend me a database with manufacturers / suppliers for a polarizing optical microscope with reflectance mode and Nomarski attachment? Any experience of a recent purchase would also be welcome.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I can't help you with your Cs value, but the incident-beam convergence semi-angle can be measured from a diffraction pattern taken with the beam adjusted as for imaging. With luck you should see a disk at each diffraction spot [1,2], and the radius of the disk is the semiangle. If you use a specimen with a known d-spacing in a known orientation, you will be able to scale the diffraction pattern in angle -- remember that 2.theta = n.lambda/d and get the semiangle in milliradian. Then you need to decide if the disks have uniforn illumination across them, or if the intensity is greater in the center. If you think that the intensity distribution is more like a 2-D Gaussian, use a HREM-simulation program that uses a Gaussian model for the convergence. If you think the disks are uniform, then use a HREM-simulation program that uses a "top-hat" model for the convergence. If the disks are uniform, but you are stuck with a HREM-simulation program that uses a Gaussian model for the convergence, make sure that the std.dev. of the Gaussian (its sigma) is set to 0.77 times the disk radius [2]. [1] "n-beam lattice images, VI. Degradation of image resolution by a combination of incident-beam divergence and spherical aberration", M.A. O'Keefe and J.V. Sanders, Acta Cryst. A31 (1975) 307-310. [2] "Using convergence and spread-of-focus parameters to model spatial and temporal coherence in HRTEM image simulations", Jan-Olle Malm and Michael A. O'Keefe, in 51st Ann Proc. MSA, Cincinnatti, Ohio (1993) 974-975. Cheers, Mike O'Keefe
Mark Blackford wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi All, } } a colleague would like to simulate HRES images to compare with those } recorded using our JEOL 2000fxII TEM. For this he needs to know the } objective lens spherical aberration coefficient (Cs). The polepiece } we have is the AHP20L. Does anyone know what the Cs value should be } for this instrument? } } Another parameter he requires is the beam semi-convergence (in mrad). } How is this parameter defined and how can we measure it? } } I would appreciate any advice you can offer. Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily } represent the official views of ANSTO from which this message was } conveyed.
http://www.amc.anl.gov/Docs/NonANL/ComSites.html#Catag } scan down the list to the light/optical/confocal listings.
howwever, this listing is NOT differentiated by the specific criteria you asked. It only represents vendors that have submitted information and is therefore not necessairly comprehensive.
Nestor
} } } } } } } } Could anyone please recommend me a database with manufacturers / suppliers } } for a polarizing optical microscope with reflectance mode and Nomarski } } attachment? Any experience of a recent purchase would also be welcome. } } } } +------------------------------------------------------------------------+ } } | Robert H.Olley Phone: | } } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } } | University of Reading {University internal extension 7867 | } } | Whiteknights Fax +44 (0) 118 9750203 | } } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } } | England URL: http://www.reading.ac.uk/~spsolley | } } +------------------------------------------------------------------------+
=========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
I'd like to get a closer look on coatings of silica-fibers using SEM. Has anybody ever tried to polish cross- and/or longitudinal sections?
Thanks for any information! Werner _______________________________________________________________________________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastrucute Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve } ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
You've addressed my concerns exactly. Then again, you're not trying to sell me a camera.
I tend to be wary of 12 megapixel output from a 1.34 megapixel chip, even if "my" math is wrong.
I too am interested in how the interpolation is done in these cameras.
It appears that some manufactures are using controlled optics to shift the image on the CCD, whilst others are shifting the CCD. How that interpolation then happens would be most informative. Here again, real info is difficult to come by on the "sales" oriented sites.Pixera seems to offer more info than others.
It would seem that by using the Bayer mask, which uses 2 green tinted pixels for 1 each of blue and red, that the resolution of the chip would decrease fourfold.
Here again, i'm just an inquiring mind that would like to know how the technology that we, the buying public, are being asked to believe.
William T. Giles Sr. Electron Microscopist Met. Lab. Coordinator Henderson Technical Laboratory TIMET PO Box 2128 Henderson NV 89009 Ph: (702)566-4436 Fax: (702)564-9038 E-mail: Bill.Giles-at-timet.com
} -----Original Message----- } From: Kuusisto, Ari [SMTP:Ari.Kuusisto-at-perkinelmer.com] } Sent: Wednesday, February 28, 2001 6:56 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Nikon DXM1200 and "mechanical" interpolation to increase the } fina l pixel count } Importance: High } } Hi all, } } I would like to ask some basic things of those cameras that scan the CCD } to } get more resolution than the specific CCD really offers. I can understand } that this can work with color CCDs that have the color mask (array or } mosaic } or whatever it is called) on the chip and cannot reach the full resolution } of the CCD. However, according to my knowledge the "scanning" cannot make } the resolution better than with the same chip used as black-and-white } camera } (without the color mask). So the scanning is done just to compensate } against } the loss of resolution caused by the color mask. Or has somebody really } invented some new rules. Could somebody please advise me, if I have missed } something here. }
I'm preparing a talk which is to include a timeline giving dates and names for some relevant TEM developments. I'm trying to find answers to two trivia questions with great importance to microscopy:
1) Who was the first to knowingly image a dislocation in a TEM, when was it done and where was it published? On this, I have found references as far back as an article by Heidenreich in J. Appl. Phys, 1949, but don't have this journal going back that far.
2) Who developed the solid state detector for energy analysis of x-rays (EDS), when and where published?
I have found some references to books on TEM history by Marton (1968) and Hawkes (1985) but don't have these on hand.
Thanks,
Wharton
*********************** Wharton Sinkler UOP LLC Des Plaines, IL
----- Original Message ----- } From: Sinkler, Wharton {WSinkler-at-uop.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 01, 2001 3:45 AM
This is a forensic microscopy question. I would like a stain(s) that differentiates between skin/protein particles and cellulose/starch particles. I would like the stain to be evident in the compound microscope under normal incident lighting. The particles are filtered from a solution of IPA and captured on a .2um polycarbonate filter. The information I have come across would suggest Ninhydrin for the staining of peptides in skin, but I have not found any info on preferentially staining cellulose/paper products.
1mm? For that I'll use a ruler. One of our stage micrometers is 0.1mm long and has 0.002mm divisions. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, March 01, 2001 2:53 AM, Seijo, Edward R. [SMTP:SeijoER-at-moffitt.usf.edu] wrote: } } } Looking for a stage micrometer that will allow me to calibrate down to the } micrometer level. Everything I have found thus far is 1mm. } } Thanks
That would be new to me. I would like to see that study. It also would be counter-intuitive since conc alcohol and acetone are known to extract lipids, whereas 50 to 70% hardly does. And since membranes are mostly lipids . . . Also during dehydration the greatest specimen shrinkage occurs above 80% alcohol. So it makes sense to have more and smaller steps at the upper end, but that is mostly due to physical effects. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, February 28, 2001 8:01 PM, Patton, David [SMTP:David.Patton-at-uwe.ac.uk] wrote: } } Re what % ethanol to use for storage. } } I once read a study which claimed 100% was better that the } usual 70%. I cannot remember the authors etc. } } Dave } } } On Tue, 27 Feb 2001 21:39:29 +1000 Jim at ProSciTech } {jim-at-proscitech.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } If those cells were for TEM sections, I would be very concerned and re- } } hydrate } } them back to 70% alcohol immediately. For SEM lipid extractions matters less } } } } but you may well get some shrinkage. Its less urgent, but I would take those } } } } cells back to 70%. At that and refrigerated you SHOULD see no real change } } for } } some weeks. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ABN: 99 724 136 560 www.proscitech.com } } } } On Tuesday, February 27, 2001 7:56 PM, Bart De Pauw } } [SMTP:Bart.DePauw-at-rug.ac.be] wrote: } } } } } } Hello Everyone, } } } } } } This week, our CPD was broken. I was planning to CPD some samples that } } } day. The samples are now in acetone. My question : how long can these } } } samples stay in acetone ? The samples are epithelial cells from duodenum } } } grown on glass. } } } } } } greetings, } } } } } } De Pauw Bart } } } Belgium } } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" }
THIRD ANNOUNCEMENT: UBC LIVE-CELL COURSE: Please register by March 1!!
(Note: I know that today is the deadline but there were still some vacancies last night and we will accept applications as long as this remains true. In addition, there is a Waiting List to replace those who find they cannot come for medical or financial reasons. JP)
Hello all,
We have two additions to the faculty for the 2000 UBC 3D Live-cell Microscopy Course:
* Alan Hibbs from BioCon in Melbourne (who was really on the list to start with but I somehow missed on the previous announcements. Sorry Alan!)
* Mark Cannell from U. Auckland NZ. Well known for developments in multiphoton microscopy but also for deconvolving 3D data from confocal and multiphoton microcopes. Mark will replace Jason Swedlow from U. Dundee who had schedule conflicts (Thanks for your help the last 3 years Jason!)
Other faculty include:
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Ted Inoué Universal Imaging, PA o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Michael Weis Agriculture Canada
Basic info is below but you can get the entire brochure, including links to faculty home pages, at
Organized by Prof. James Pawley, (University of Wisconsin-Madison)
in association with Dr. Elaine Humphrey UBC BioSciences Microscopy Facility: University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 1, 2001 Deposit due April 15, 2001 Registration 5:00 - 7:00 PM Sunday, June 17, 2001 First Lecture 7:30 PM Sunday, June 17, 2001 Live-cell Course ends, noon Thursday, June 28, 2001 3D Image Processing Wksp Sat. June 30 - Monday, July 2
APPLICATIONS DUE BY MARCH 1, 2001
More information at : http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
or
REGARDING COURSE ORGANIZATION:
Contact: Prof. James B. Pawley, JBPAWLEY-at-FACSTAFF.WISC.EDU
REGARDING APPLICATIONS
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, an intensive eleven-day residential course concentrating on all aspects of 3D Microscopy of Living Cells will be held at the University of British Columbia, in June of 2001. The course includes 4 days on 2D techniques, 6 days of 3D techniques and a summary presentation. It covers basic microscopy to the highest level confocal microscopy. (A half-day Pre-course is offered for any who may need to brush up on basic optics!).
Topics include: o Quantitative 2D light microscopy o How to keep your cells alive o 3D imaging in confocal microscopy o Widefield/deconvolution techniques o Two-photon excitation microscopy o Fluorescent and backscattered light signals o Dye design, characteristics and use o Pixelation: The Nyquist Criterion o Lasers and laser tweezers o Objectives and aberrations o Scanning-systems: AODs and mirrors o Optimal pinhole size/photon efficiency o Detectors: operation and performance o Video-rate confocal imaging o Measuring ion concentrations o Display and measurement of 3D data
Morning lecture/demonstrations lead to hands-on laboratory exercises each afternoon that will utilize most of the commercial instruments currently available for 3D microscopic imaging. Students will work in groups of 3 or 4 throughout the discussion and laboratory sessions, and may complete a live-cell 3D study on their own specimens. In the first five years, over 130 students from 23 countries have attended. Last year, 11 separate, 3D microscopical workstations were available for student use under the supervision of a 17-member international faculty. We expect to have even more workstations in 2001. Including manufacturers representatives, the teacher/ student ratio will be almost 2:1.
INTERNATIONAL FACULTY
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o mark Cannell Univ. of Auckland o Elaine Humphrey Univ. of British Columbia o Alan Hibbs BioCon, Melbourne o Stephan Hell Max Planck, Goettingen o Elaine Humphrey University of British Columbia o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Glen MacDonald University of Washington o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Michael Weis Agriculture Canada
TUITION
Course tuition is $2,150 US and includes lunches. On receipt of 50% deposit, students will receive preliminary group assignments and the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes the textbook, a binder of handouts, and tickets for the Opening Reception, the Manufacturer's Reception and the Beach Party. Accommodations and meals other than lunch are not included in the tuition fee. The Pre-course is $100 US.
APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrollment is limited to about 24 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts on request to read before the course begins and are encouraged to take the Pre-course on the afternoon of June 18.
Application forms, and other course information from this and past years, can be downloaded from the WWW site at:
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4 Phone: 1-604.822-3354 FAX: 1-604.265-5315 Email: ech-at-unixg.ubc.ca.
Application deadlines:
Application forms are due by March 1, 2000! Deadlines are early to facilitate setting up groups. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2000 to reserve your position. In general, deposit refunds are only be possible if your position can be filled from the waiting list. The remainder of the fees is due at registration.
DATES
Applications must be received by March 1, 2001 Deposit due April 15 2001 Registration 5:00 - 7:00 PM Sunday, June 17, 2001 First Lecture 7:30 PM Sunday, June 17, 2001 Live-cell Course ends, noon Thursday, June 28, 2001
TEACHING PHILOSOPHY
As befits teaching in an area at the boundary of "what is now known," lecturers have been chosen based on their expertise as scientists working in the field rather than because they all agree. They are encouraged more to be provocative than to be prosaic. Students should expect discussion in areas where differences of opinion exist.
Prior to the course, students will be organized into groups and encouraged to communicate by email/phone, about the "Living-cell" group projects that they will pursue during the course and that will be presented to the class on the last day of the course. It has been found that group interactions make best use of students' prior experience and can be very effective in teaching the practical skills covered in a hands-on course of this type.
ARRANGEMENTS FOR LIVE SAMPLES
Students must contact Dr. Elaine Humphrey to make necessary arrangements for the transport and maintenance of cell lines etc. needed for their projects. Organisms linked in any way with human disease are not permitted because of safety considerations. Transport and customs arrangements for living specimens are entirely the responsibility of the student. Students also attending the 3D Image Processing Course, may be able to analyze, process and display some of the 3D data collected from their specimens
The workshop will cover 3D image processing for measurement and display. Enrollment is limited to those attending the 3D Microscopy course. Tuition : $850 US (lunches and snacks incl.)
WHO SHOULD ATTEND?
The course is designed for biologists working with multidimensional and possibly multicolor microscopical data sets. Getting the data is only half of the battle. Image data in 3, 4 or even 5 dimensions may be difficult to store let alone analyze or display. This course is to help students understand the hardware and software aspects of this problem and give them the techniques they need to make the best use of their data.
The course is designed for biologists who need to make measurements on 3D microscopical data sets and then display the results in an effective manner. The course will be taught on SGI, Macintosh and IBM-compatible computers. A wide variety of software designed for the 3D microscopy market will be described, demonstrated and available for use.
Workshop Organizers
o Andreas Kriete Giessen University, DE o Felix Margadant University of Sydney, Au o Ping Chin Cheng State U. of New York, Buffalo o Elaine Humphrey University of British Columbia o Glen MacDonald University of Washington
PLAN OF INSTRUCTION Classes will meet from 8:30-12:00 and 1:00-6:00 with lecture-demonstrations followed immediately by hands-on laboratory sessions using a variety of workstations. Students will "learn-by-doing" with two to a machine. Lab handouts will describe some specific exercises to be performed on "canned" data sets. Facilities and supervision will be available until 11:00 PM, for students to work on their own data. -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
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} I have found some references to books on TEM history by Marton (1968) } and Hawkes (1985) but don't have these on hand.
I have a book that was published at the time of the International Congress on EM in Kyoto, Japan, in 1986, entitled "History of Electron Microscopes 1986" edited by Hiroshi Fujita. This gives a lot of information on the history of EM, mainly from a Japanese perspective. I haven't seen whether or not this has the answers to your specific questions but if there is someone near you who has a copy of this book you will find much interesting information in it. Please let me know if you cannot get hold of a copy.
Robin H Cross (Mr) Director : Electron Microscopy Unit Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/index.htm
** remember ICEM-15 in Durban in September 2002 **
I am not a forensic scientist so my suggestions are general - therefore I am uncertain how the methods I give could be made robust enough for use in court. I assume that IPA is isopropyl alcohol?
Starch should be positive with the periodic acid Schiff method (PAS) and may be 'confirmed' as such by using an amylase control. Starch should also show 'Maltese Cross' birefringence under polarised light. The cellulose should also be PAS positive, resist amylase and show 'linear' birefringence.
The protein is a little more difficult to confirm absolutely but if these are skin cells they will contain cytokeratins - thus giving the possibility of using immunocytochemistry with a pancytokeratin antibody.
Good luck!
PS Why is an IR/Raman engineer doing forensic work on paper/cells?
At 16:07 2001-02-28 -0800, HAMMOND,LOMA (HP-Corvallis,ex1) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
Congo red will stain cellulose red, and can also be viewed by fluorescence using a green (RITC-type) excitation filter. Cellulose and starch are also birefringent, which you could demonstrate by using crossed polarizers. A 1/4 wave or 1-wave plate may make this easier to see. The fluorescence of Cellulose stained with congo red is also polarized because the CR molecules line up on the oriented cellulose microfibrils. As has already been mentioned starch and cellulose stain with periodate-Schiff. If you have access to a fluorescence microsocope you might also try a fluorescence variant of PAS, by replacing schiff reagent with Lucifer Yellow CH. View with FITC-type excitation.
Iodine/potassium iodide will stain starch blue/black.
Chris
----- Original Message ----- } From: "HAMMOND,LOMA (HP-Corvallis,ex1)" {loma_hammond-at-hp.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 01, 2001 12:07 AM
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Fungal people, Although vapor fixation works sometimes and immersion fixation also works sometimes, often fungi samples are too delicate to handle with routine techniques. Just immersion in an aqueous solution can damage the structures. Repeated washings also can wash away delicate conidia. CPD causes shrinkage which may not be a major concern with some samples but can be a real problem with delicate ones like fungal hyphae. Really the very best way to deal with many fungi samples is cryo SEM. True you may have to coat a bit longer than normal, etc. but the structures, when instantly frozen, are most likely to be similar to the living state. Unfortunately many people do not have access to Cryo -SEM equipment so must settle for the next best approach....fixation and drying by one of the methods already mentioned.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
Microlisters,
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastrucute Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve } ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
As others have pointed out, phase halo is easily demonstrated. Shading-off is a different matter, at least for me. Both artifacts are the result of incomplete segregation of the direct and diffracted light to the conjugate and complementary areas respectively of the phase plate. Shading-off is described in "Advanced Light Microscopy" vol. 2 pg. 15-20 by Maksymilian Pluta. Shading-off appears as a brighter central area becoming darker toward the edge in positive phase contrast, and just the reverse for negative phase contrast. Pluta has illustrations of this on pages 18 and 19, but he doesn't say what the object is. It looks like a crystal of some kind.
My point in bringing this up is to solicit others ideas on how best to teach students (as well as ourselves) to interpret the phase contrast image. The phase image has these artifacts superimposed on it. The phase halo can mislead a novice but actually help an expert. Shading-off is more subtle in a complex specimen than it is in Pluta's example, and I have never seriously considered how it affects the phase image. I wonder if there is a biological sample that clearly shows both artifacts? Any thoughts and suggestions on this?
Bob
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718 http://www.pathology.med.unc.edu/path/microscopy/welcome.htm
Email: kuznetsv-at-geo.komisc.ru Name: Chuprov Georgy School: The Institute of Geology State: Republic of Komi
Question: Please, tell me. Where I can find a computer program for plot of stereografic projections of optical axes (were tested by optical microscop)?
I have one of those very basic "thought" questions on a procedure that I've always taken for granted, but am now not so sure about. I had been trained from Day One to always focus a TEM with the beam at crossover, then to spread the beam until the exposure intensity is reached. The idea seems to be that small focusing errors at crossover will be corrected as the beam is spread and "depth of focus" is increased somehow.
The reason I'm asking is that in recent years, almost no one else I've run across focuses in this way, preferring instead to focus with the beam already spread to the point at which the photograph will be taken (at least as long as the image is bright enough to see for focusing). Out of curiosity, I ran tests using both methods and have noted no clear differences.
Does anyone have any thoughts on this? Any ideas or theories about why one method would be superior to the other?
Sincerely, Curious in Columbia
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Dr. Ahlstrand's technique is excellent for imaging hyphal structures and some spore bodies while the stick on method of Dr. Mary Mager works well for certain individual spores. However, imaging of fruiting body structures (conidiophores, etc.) are often best prepared by using Osmium vapor fixation followed simply by air-drying (AD). One can also add minimal solvent exchange before AD, but some structural collapse will be observed in any event. "One size does not fit all" for fungi, sometimes even within one species.
We have a new ESEM now and hopefully combinations of low vacuum technology and various wet modes will improve imaging for our mycologists.
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Microlisters,
While the simpler and less time consuming suggestions already mentioned in this thread may work in some cases, I found out a long time ago that the only way to deal with the "fungal jungle", eg. fungi cultured on agar plates, is to hit 'em with the ol' OTO method, preceded by glutaraldehye fixation. Quick outline is as follows:
Glut. fixation, your favorite buffer Buffer rinses Osmium tetroxide, 2% Water rinses Thiocarbohydrazide, sat. soln.,filtered water rinses Osmium tetroxide, 2% Water rinses dehydration series, EtOH, or acetone CPD metal coating? Maybe, maybe not. Try without first.
This eliminates the charging, and fixation is quite good.
Classic reference for this method:
"Ligand-mediated osmium binding:Its application in coating biological specimens for scanning electron microscopy". Kelley, Robert O., A.F. Dekker and John G. Bluemink. J. Ultrastructure Research 45:254-258 (1973).
See also:
"Non-coating techniques to render biological specimens conductive/1980 update". Judith A. Murphy. Scanning Electron Microscopy/1980 Vol.I, p. 209-220.
You might try exposing your non-aquatic sample (bread molds, mushrooms, etc. ) to vapors of Osmium tetroxide for several hours ( a few drops of 4% OsO4 on the lid of a very small container with your sample inside or the lid over the sample directly), then air dry, mount, coat and view. As with any sample, the drying artifacts vary with the fungus. Some things look great.
good luck
Steve -----------------------------------------------------------------
Dear rad0, I have looked at fungus and fungus spores on my SEM and they are very simple to prepare. Just stick them to a stub by sticky tab or glue and then gold coat. They are very robust and they don't need any fixing or dehydration.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} ----------------------------------------------------------------- } Hello, } } Forgive me, I haven't tried to find this on my own yet... } } But have any of you used an electron microscope to get a } close-up of a fungus? Can this be done? } } Thanks...
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
You might guess from the lack of response that quantitative stereoscopy using TEM is not that popular a pastime. I was involved some 20 years ago and can offer some comments which were relevant then.
There is a big difference between TEM and SEM stereo analysis. In SEM images you have a lower surface that can be seen easily, in TEM images you have a view through both surfaces and the specimen, the surfaces are very hard to distinguish. It is relatively easy to calculate the height from a fixed surface as in a SEM micrograph. Calculating the height from an 'invisible' surface, as in a TEM micrograph, is very difficult. It's OK to think of something like a dislocation running through the specimen and then assume that it ends at either surface but very few examples go through from surface to surface and thus are considerably more difficult.
Height measurements are made from stereo pairs by measuring the parallax between the two tilted images. Provided the tilt axis is known and the magnifications corrected, for any change in focus current, then the height difference between two points can be calculated. This is a long way from reconstructing a 3D image.
It is relatively easy to view stereo images using a stereo viewer but not everyone can do it. You need two eyes of equal strength and a certain type of brain. From memory the mapmakers said 50% of the population could see stereo properly and 10% could make accurate measurements.
There were programs that would allow 2 stereo micrographs to be placed side by side on a graphics tablet and then the same points marked on both micrographs. The height differences between these points were then calculated using the first pair of points as a reference height. I don't know if these are still available for modern computers (they ran on an Apple 11+).
Stereo viewers are available that will project a 'floating' point of light in the specimen that you can position onto a feature and then read out the height from a micrometer (it will need to be scaled to get the real height). We modified one of these viewers to record and calculate these heights (and X,Y coordinates) automatically), but it it still a lot of work to produce even a simple 3D image.
I am not aware of any image recognition program that will analyse your two images and produce a 3D image from them. (Well if that doesn't get details sent in nothing will!)
If you want some references then in the mid 70s Hudson and Makin descibed the how to select the tilt angle taking into account the film thickness and the final magnification. Much of the work at that time was being done by Alan Boyde. I may be more use with references when I get my office back - contact me off list in a few days if you want 20-25 year old refences.
Good luck, Ron
On Wed, 28 Feb 2001 10:03:56 -0700 Rodney McCabe {rmccabe-at-lanl.gov} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues: } } I am interested in doing quantitative stereo microscopy using a TEM and am } looking for software that may be helpful in such an endeavor. I anticipate } the primary application will be examining the spatial arrangement of linear } defects. I have not been able to find a software designed directly for } quantitative stereo TEM. I have come across one commercial software that } can be used for SEM surface topography that may be applicable. It seems } like there must be other software packages out there used for applications } such as surface mapping, electron tomography, or even astronomy that could } be useful. Also, as I am new to stereo microscopy, any words of wisdom } would be appreciated. } } Thanks, } } Rod } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
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Pete Augustus Caswell Technology
Hi Listers,
I'm preparing a talk which is to include a timeline giving dates and names for some relevant TEM developments. I'm trying to find answers to two trivia questions with great importance to microscopy:
1) Who was the first to knowingly image a dislocation in a TEM, when was it done and where was it published? On this, I have found references as far back as an article by Heidenreich in J. Appl. Phys, 1949, but don't have this journal going back that far.
2) Who developed the solid state detector for energy analysis of x-rays (EDS), when and where published?
I have found some references to books on TEM history by Marton (1968) and Hawkes (1985) but don't have these on hand.
Thanks,
Wharton
*********************** Wharton Sinkler UOP LLC Des Plaines, IL
For fragile conidiophores and their associated conidial chains, it is still possible to use critical point drying after vapor fixation with OsO4.
A fairly simple dehydration technique that avoids the solution changes -- and turbulence -- that often lead to loss of the conidia from aerial conidiophores (and the air drying that often does the same), is described at Can. J. Microbiol. 29:653-658 (1983).
It's an adaptation of a slick idea published in Naturwissenshaften 17:402-403, for TEM, in 1962(!).
I always "heard" to focus the image with the beam in an overfocus condition (past crossover) for improved beam coherence. I find it difficult to focus at crossover anyway. Also, I don't think you can accurately assess focus or astigmatism with the beam at intensities suitable for most photography. Depends on the magnification and a host of other things, of course, such as your exposure time. I don't think there's any justification to focus at exactly the same intensity at which you will photograph. I too await further enlightenment from the list....
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } I have one of those very basic "thought" questions on a procedure that I've } always taken for granted, but am now not so sure about. I had been trained } from Day One to always focus a TEM with the beam at crossover, then to } spread the beam until the exposure intensity is reached. The idea seems to } be that small focusing errors at crossover will be corrected as the beam is } spread and "depth of focus" is increased somehow. } } The reason I'm asking is that in recent years, almost no one else I've run } across focuses in this way, preferring instead to focus with the beam } already spread to the point at which the photograph will be taken (at least } as long as the image is bright enough to see for focusing). Out of } curiosity, I ran tests using both methods and have noted no clear } differences. } } Does anyone have any thoughts on this? Any ideas or theories about why one } method would be superior to the other? } } Sincerely, } Curious in Columbia } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } }
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC) Materials Characterization Facility (MCF)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for up to two positions as Assistant in Research to support the Materials Characterization Facility (MCF). Each individual must have at least a Bachelor degree from an accredited institution in an appropriate area of study related to surface science, materials characterization, ion beam analysis, or vacuum science, and preferably three years of experience in any of the above mentioned areas. A graduate degree in the aforementioned areas is preferred. The MCF houses an RBS, a heavy ion beam system, an Auger, an XPS, two FIBs and four SIMS instruments. Opportunities also exist with collaborative interaction with Lucent Technologies, Orlando, which houses an Auger, four FIB systems including a FIB/SIMS and two dedicated SIMS tools.
The person will be responsible for establishing, maintaining and operating ion beam and/or surface science laboratory infrastructure. Depending on the experience, the person should be able to instruct students, faculty, and staff on the use of equipment and in the interpretation of data, conduct independent research and develop surface analysis techniques. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. Salary will be commensurate with experience. The applications will be reviewed beginning March 19, 2001 and will continue to be reviewed until the position is filled.
Applicants should send a vitae and a list of three references to Dr. Lucille Giannuzzi, Director MCF, 12443 Research Parkway, Suite 305, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
I have focused at a higher beam intensity than I use for exposing the micrograph for many years, and, in fact continue to do so on our Philips EM201. When we got a new Philips TwinLens CM12S in December of 1988, we started to have problems with slightly out of focus micrographs across the board - all experienced biological microscopists - about 10-12 of us - had the same problem. Philips, to their credit, tried repeatedly to find and solve the problem. They sent high level engineers, applications experts, attached a high mag TV camera, changed all of the high tension cabling, insulator, wehnelt, column liner, etc. over the course of nearly a year. Towards the end of 1989, in desperation and because the biological TEM applications experts were all busy, a materials science applications engineer came to the lab and made beautiful, perfectly focused micrographs. We could not duplicate his performance. Finally, he asked to watch one of our most experienced and well published microscopists work. When the Professor increased beam intensity to focus and stigmate, the applications engineer went ballistic! he maintained that the focus MUST be done at the intensity at which the micrograph is made. We did argue, but also agreed to test the hypothesis. He was, of course, correct (IN THIS PARTICULAR CASE). As we later found out, changing the C2 lens current also has a subtle effect on the objective lens current, changing focus enough to cause problems. Our service engineer was later able to demonstrate the effect (although it doesn't show up when monitoring the lens current page) and this strange phenomenon was written off to a design compromise which, apparently, physicists and materials scientists are used to and expect, but which biologists and cell biologists had never heard of. Something to do with short focal length objective lenses, low contrast which makes focusing more difficult, and compromises required to allow STEM and TEM to co-exist in the same column and be switched back and forth with a minimum of re-alignment. Anyone else have this experience?? William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
I spread the beam as much as possible in order to be able to see the changes during the focusing (usually the intensity is less than necessary to make a photograph, so, I have to condense the beam a little bit for photographing). I am using "three-chick" technique: rotating focus-control knob in both directions to find the position when you will clearly see the transition: "owerfocus-infocus-underfocus" on the background's granules. This technique works for me from x40K and up. At the lower magnification, I am using wobbler to focus (there is a "blind spot" at x30-40K). The "three-chick" technique is a good test how your instrument is aligned. The number of "clicks" depends from current setup and model of coarse. I simply could not focus correctly if the beam is condensed too much.
Sergey
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Well, I got my answer----right from one of the people who initially trained me back in the '80's (and, in fact, is responsible for me being in the field in the first place----that'll teach him).
Here is John Bozzola's reply to my post on focusing TEM's at beam crossover versus a defocused beam: "(T)his is a holdover from (our Hitachi) HU11A when illumination was a problem at the higher mags and when no focus wobbler was available for low mag work. On the other hand, it still DOES work for individuals who have diminishing eyesight and need the extra illumination and minimal depth of field. Unfortunately, it greatly increases the possibility of specimen damage and should be used only on hardy specimens.
"The bottom line is that if you have normal (or acute) vision, you won't need to do this in modern TEMs. Of course, I sometimes still "check" my focus by doing this..... old habit, I guess."
Now this is very interesting to me, because the first TEM I ever used was the Hitachi HU11AB. I was trained to focus carefully at crossover, then defocus until the proper exposure intensity was reached. Somehow over the years, I ended up with the notion that there was a property of electron "optics" that made this the preferred method of focusing because it minimized focusing errors by increasing the depth of focus. I really don't recall where this notion came from, but I trained dozens of students and others in this way of focusing. Now I wonder how many others were trained by them to do the same thing and will be just as stubborn about it as I have been.
Probably not much harm done, really, but this reminds me of one of my favorite stories about a woman cooking a ham for a family reunion picnic. Her daughter watched her mother cut off the end of the ham before putting it in a pan in the oven, and she asked why she did that. Her mother responded that that's how she was taught by HER mother, but she didn't really know why. She became curious, and at the reunion she asked her mother why she had always cut the ends off hams before cooking them. The response was that she had also been taught that way by her mother. Both curious now, they hunted up Great-grandmother at the picnic and asked her why she had cooked hams with the ends cut off. Great-grandmother answered, "I only had one pan and it was too short for the ham."
Lesson learned. Again.
Cheers, Randy
Randy Tindall Electron Microscopy Core University of Missouri
In terms of contrast transfer theory (i.e. when linear imaging allows approximating the imaging process as convolution with a point spread function), the lens transfer functon doesn't depend on convergence. So for example you shouldn't effect the defocus of the image by slight changes of the illumination convergence.
However, the convergence imposes a damping on the total contrast transfer, which gets more severe when convergence is greater. For details on this 'spatial coherence envelope' see any text on HREM. Because of this, the image quality will improve if you record with a less condensed beam (better coherence).
With a field emission gun, you won't be able to image at crossover, because the source will almost certainly be too small - these guns give the most coherent illumination available.
For a thermionic gun, the limiting factor will be: if the convergence is too great at crossover, you won't see much because everything will be blurred out by the coherence limitation of the incident irradiation. If this is the case, you should either change condenser apertures to decrease convergence (then you can focus, and/or stigmate at crossover), or spread the beam to improve coherence.
As far as I can see, there is no reason why one should or should not adjust or even record images at crossover (except that if the filament is a bit undersaturated, you will get uneven illumination). Getting the best possible coherence is important, but it doesn't really matter how one achieves this optically. Decreasing intensity is therefore good, to the extent that sample drift doesn't become too much a limiting factor.
Overfocusing the illumination rather than underfocusing is better, I would say (based on experience). I'm not sure why - perhaps some of the energy spread gets filtered out more effectively - at any rate for the same size beam one has more intensity underfocused than overfocused, so something is being cut out by overfocusing.
Wharton
} -----Original Message----- } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } Sent: Thursday, March 01, 2001 2:30 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: Focusing } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I always "heard" to focus the image with the beam in an overfocus } condition } (past crossover) for improved beam coherence. I find it difficult to } focus at } crossover anyway. Also, I don't think you can accurately assess focus or } astigmatism with the beam at intensities suitable for most photography. } Depends on the magnification and a host of other things, of course, such } as } your exposure time. I don't think there's any justification to focus at } exactly the same intensity at which you will photograph. I too await } further } enlightenment from the list.... } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } [SMTP:TindallR-at-missouri.edu] wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have one of those very basic "thought" questions on a procedure that } I've } } always taken for granted, but am now not so sure about. I had been } trained } } from Day One to always focus a TEM with the beam at crossover, then to } } spread the beam until the exposure intensity is reached. The idea seems } to } } be that small focusing errors at crossover will be corrected as the beam } is } } spread and "depth of focus" is increased somehow. } } } } The reason I'm asking is that in recent years, almost no one else I've } run } } across focuses in this way, preferring instead to focus with the beam } } already spread to the point at which the photograph will be taken (at } least } } as long as the image is bright enough to see for focusing). Out of } } curiosity, I ran tests using both methods and have noted no clear } } differences. } } } } Does anyone have any thoughts on this? Any ideas or theories about why } one } } method would be superior to the other? } } } } Sincerely, } } Curious in Columbia } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } }
We're analysing Cu wire here for purposes better known to ourselves. We are noticing, but not able to image interesting variations in the concentration of impurities, which suggests to us, either that the original material was inhomogeneous, or that drawing the wire has caused some kind of exsolution process. Have any of you encountered smilar things in metals? Refs? Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
To add to Wharton's explanation the reason overfocus is better than underfocus is that he has a strong objective lens (as most are these days). Many years ago underfocus would have been better.
What we are trying to achieve is parallel illumination not convergent (or divergent). Modern TEMs have strong objective lenses and use the prefield (field above the specimen) as part of their probe forming optics thus if there is a crossover above the specimen (overfocus)the prefield converges the diverging rays from this to form a parallel probe. In underfocus conditions the crossover is below the specimen and the probe on the specimen is made more convergent by the objective lens prefield.
If you have a Lorentz (low field for magnetic work) pole piece or an old instrument (pre 1970ish) the objective lens is weak and does not have this prefield effect so underfocus is more parallel.
Regards, Ron
On Thu, 1 Mar 2001 15:30:22 -0600 "Sinkler, Wharton" {WSinkler-at-uop.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } In terms of contrast transfer theory (i.e. when linear imaging allows } approximating the imaging process as convolution with a point spread } function), the lens transfer functon doesn't depend on convergence. So for } example you shouldn't effect the defocus of the image by slight changes of } the illumination convergence. } } However, the convergence imposes a damping on the total contrast transfer, } which gets more severe when convergence is greater. For details on this } 'spatial coherence envelope' see any text on HREM. Because of this, the } image quality will improve if you record with a less condensed beam (better } coherence). } } With a field emission gun, you won't be able to image at crossover, because } the source will almost certainly be too small - these guns give the most } coherent illumination available. } } For a thermionic gun, the limiting factor will be: if the convergence is too } great at crossover, you won't see much because everything will be blurred } out by the coherence limitation of the incident irradiation. If this is the } case, you should either change condenser apertures to decrease convergence } (then you can focus, and/or stigmate at crossover), or spread the beam to } improve coherence. } } As far as I can see, there is no reason why one should or should not adjust } or even record images at crossover (except that if the filament is a bit } undersaturated, you will get uneven illumination). Getting the best } possible coherence is important, but it doesn't really matter how one } achieves this optically. Decreasing intensity is therefore good, to the } extent that sample drift doesn't become too much a limiting factor. } } Overfocusing the illumination rather than underfocusing is better, I would } say (based on experience). I'm not sure why - perhaps some of the energy } spread gets filtered out more effectively - at any rate for the same size } beam one has more intensity underfocused than overfocused, so something is } being cut out by overfocusing. } } Wharton } } } } -----Original Message----- } } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } } Sent: Thursday, March 01, 2001 2:30 PM } } To: 'Microscopy-at-MSA.Microscopy.com' } } Subject: RE: Focusing } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I always "heard" to focus the image with the beam in an overfocus } } condition } } (past crossover) for improved beam coherence. I find it difficult to } } focus at } } crossover anyway. Also, I don't think you can accurately assess focus or } } astigmatism with the beam at intensities suitable for most photography. } } Depends on the magnification and a host of other things, of course, such } } as } } your exposure time. I don't think there's any justification to focus at } } exactly the same intensity at which you will photograph. I too await } } further } } enlightenment from the list.... } } } } Matt } } } } Matthew J. Lynn } } Center for Advanced Microscopy } } University of Miami } } (305)284-4736 } } mlynn-at-miami.edu } } } } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } } [SMTP:TindallR-at-missouri.edu] wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi all, } } } } } } I have one of those very basic "thought" questions on a procedure that } } I've } } } always taken for granted, but am now not so sure about. I had been } } trained } } } from Day One to always focus a TEM with the beam at crossover, then to } } } spread the beam until the exposure intensity is reached. The idea seems } } to } } } be that small focusing errors at crossover will be corrected as the beam } } is } } } spread and "depth of focus" is increased somehow. } } } } } } The reason I'm asking is that in recent years, almost no one else I've } } run } } } across focuses in this way, preferring instead to focus with the beam } } } already spread to the point at which the photograph will be taken (at } } least } } } as long as the image is bright enough to see for focusing). Out of } } } curiosity, I ran tests using both methods and have noted no clear } } } differences. } } } } } } Does anyone have any thoughts on this? Any ideas or theories about why } } one } } } method would be superior to the other? } } } } } } Sincerely, } } } Curious in Columbia } } } } } } Randy Tindall } } } EM Specialist } } } Electron Microscopy Core Facility } } } W122 Veterinary Medicine } } } University of Missouri } } } Columbia, MO 65211 } } } Tel: (573) 882-8304 } } } Fax: (573) 884-5414 } } } Email: tindallr-at-missouri.edu } } } Web: http://www.biotech.missouri.edu/emc/ } } } } } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
we are looking for a working Nanoscope III (III or IIIa) controller together with the PC boards (Digital Instruments, Santa Barbara, USA). If somebody is going to clean out its lab please contact us directly by phone, fax or e.mail
With best regards,
Dr. Dmitry Cherny, PhD, Dr.Sc.
MPI for Biophysical Chemistry, Dept. of Molecular Biology am Fassberg 11, D-37077 Gottingen, Germany tel: +49(0) 551 201 1383; fax: +49(0) 551 201 1467 e.mail: dtcherny-at-mpc186.mpibpc.gwdg.de
does anyone have any suggestions on how to assess hydronephrosis with paraffin sections of kidney using a quantitative technique. equipment we currently use for other tissues include microscope, spot camera, digitizing tablet, camera lucida, SigmaScan, and image processing software plugins for adobe photoshop,
Nonuniform distribution of impurities in metal wires is not uncommon. There may be a core-to-surface variation, which may be caused by contamination (or de-contamination) of the surface layer during drawing or annealing . Variations in impurity concentration may also relate to original inhomogeneity in the billet prior to drawing. Segregation of impurities to dislocation-rich deformed areas is also a possibility.
What are the impurity elements that you are seeing? How are the concentration variations distributed spatially? (Radial gradients, irregular patches smaller than wire diameter or larger than wire diameter, wire bends vs. straight segments, or what?)
-------Roy ==================================== Roy Arrowood, Associate Professor Metallurgical and Materials Engineering UTEP, El Paso, TX 79968-0520 (915)747-6934 arrowood-at-utep.edu
} From a practical standpoint, wouldn't it be best to focus using the same conditions that are later used for taking the images, unless there are intensity or other issues that prevent that?
A few years back, when I was doing hi-res TEM, we would carefully set the conditions on the TV screen, then turn off the room fan, move away from the column, hold our breath and take a picture.
However, this may have been peculiar to hi-res materials work, where even tiny changes in focus can have dramatic effects.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Thursday, March 01, 2001 2:30 PM To: 'mlynn-at-miami.edu'; 'Microscopy-at-MSA.Microscopy.com'
In terms of contrast transfer theory (i.e. when linear imaging allows approximating the imaging process as convolution with a point spread function), the lens transfer functon doesn't depend on convergence. So for example you shouldn't effect the defocus of the image by slight changes of the illumination convergence.
However, the convergence imposes a damping on the total contrast transfer, which gets more severe when convergence is greater. For details on this 'spatial coherence envelope' see any text on HREM. Because of this, the image quality will improve if you record with a less condensed beam (better coherence).
With a field emission gun, you won't be able to image at crossover, because the source will almost certainly be too small - these guns give the most coherent illumination available.
For a thermionic gun, the limiting factor will be: if the convergence is too great at crossover, you won't see much because everything will be blurred out by the coherence limitation of the incident irradiation. If this is the case, you should either change condenser apertures to decrease convergence (then you can focus, and/or stigmate at crossover), or spread the beam to improve coherence.
As far as I can see, there is no reason why one should or should not adjust or even record images at crossover (except that if the filament is a bit undersaturated, you will get uneven illumination). Getting the best possible coherence is important, but it doesn't really matter how one achieves this optically. Decreasing intensity is therefore good, to the extent that sample drift doesn't become too much a limiting factor.
Overfocusing the illumination rather than underfocusing is better, I would say (based on experience). I'm not sure why - perhaps some of the energy spread gets filtered out more effectively - at any rate for the same size beam one has more intensity underfocused than overfocused, so something is being cut out by overfocusing.
Wharton
} -----Original Message----- } From: Matthew Lynn [SMTP:mlynn-at-miami.edu] } Sent: Thursday, March 01, 2001 2:30 PM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: Focusing } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I always "heard" to focus the image with the beam in an overfocus } condition } (past crossover) for improved beam coherence. I find it difficult to } focus at } crossover anyway. Also, I don't think you can accurately assess focus or } astigmatism with the beam at intensities suitable for most photography. } Depends on the magnification and a host of other things, of course, such } as } your exposure time. I don't think there's any justification to focus at } exactly the same intensity at which you will photograph. I too await } further } enlightenment from the list.... } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Thursday, March 01, 2001 10:28 AM, Tindall, Randy D. } [SMTP:TindallR-at-missouri.edu] wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi all, } } } } I have one of those very basic "thought" questions on a procedure that } I've } } always taken for granted, but am now not so sure about. I had been } trained } } from Day One to always focus a TEM with the beam at crossover, then to } } spread the beam until the exposure intensity is reached. The idea seems } to } } be that small focusing errors at crossover will be corrected as the beam } is } } spread and "depth of focus" is increased somehow. } } } } The reason I'm asking is that in recent years, almost no one else I've } run } } across focuses in this way, preferring instead to focus with the beam } } already spread to the point at which the photograph will be taken (at } least } } as long as the image is bright enough to see for focusing). Out of } } curiosity, I ran tests using both methods and have noted no clear } } differences. } } } } Does anyone have any thoughts on this? Any ideas or theories about why } one } } method would be superior to the other? } } } } Sincerely, } } Curious in Columbia } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } }
*************************************************** PROGAM: FOCUS ON MICROSCOPY 2001 ***************************************************
Dear colleagues,
The program of the conference "Focus on Microscopy 2001", April 1 - 4, 2001, Amsterdam, has been finalised and can be found on our Web-site
www.focusonmicroscopy.org
This conference is the 14th in a successful series of interdisciplinary meetings on 3D acquisition and 3D image processing and forms an effective meeting point for both developers and users of 3D-microscopy. The more than 120 contributions this year range from new developments in Coherent Anti-Stokes Raman Microscopy (CARS) and multi-photon excitation to the application of such techniques in biology, medicine and material sciences. This year "3D Microscopy of living cells and tissues" will receive special attention, which reflects the growing number of researchers that use GFP-techniques and life-cell imaging for their experiments.
A substantial instrument exposition including the main manufacturers in the field will accompany the conference.
Please, visit our web site http://www.focusonmicroscopy.org for further details concerning the program and the conference.
Looking forward to seeing you in 1st of April in Amsterdam, on behalf of the program committee,
We are working with embbeded cells in LR White with and without osmium. (Polimerization was acomplished at 60ºC). Our purpose is to detect gangliosides (GD3). Do you have any suggestion?
I also was told early on in my career that one should focus at the magnification the picture was to be taken at, though I find it difficult to do this in practice because the illumination is often too low. I usually compromise and focus with as little illumination as I can get away with.
I wonder whether the reason for the change in focus is not due to changes in the illumination, but changes in the specimen position. At different beam densities the degree of specimen charging may vary, thereby changing the position of the specimen in the lens field. This might explain why some microscopes are less sensitive to the specimen current density than others. Certainly the amount of specimen charging seems to be affected by the objective aperture, for example.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Peggy: I don't know if you received any responses off-line, and I hope that Ronald Austin was able to provide you with a protocol for his microwave procedure (actually, if you have one available Ronald, I'd like a copy, please). Your question is most appropriate because that is what I am (frantically) in the midst of doing. Altho' "frantic" applies only to the deadline.... Sans a microwave procedure, I am doing the old time-tested methods. The only thing I make sure of is the buffer used to transfer the tissue from formalin. I add 6% sucrose to the buffers and carry that through until I get into osmium. Loss of membranes occurs in the formalin, and if you add the 6% sucrose it seems to help retain membranes in the final EM sample.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On 26 Feb 2001 15:06:20 -0500 (EST), "HARGER-ALLEN.MARGARET_J-at-INDIANAPOLIS.VA.GOV"-at-sparc5.microscopy.com wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Pathology lab. listers: are there any new/and/or superior techniques | for processing previously formalin fixed tissue for TEM? | Thanks, Peggy Harger-Allen | |TAB|email:harger-allen-at-indianapolis.va.gov |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
The pellets are from a hydrobiid snail found in highly mineralized warm desert springs and Posos in Mexico. I am interested primarily in the bacteria found on and in them but would also like to be able to inventory the entire contents of the pellets to help get a better picture of what part they play in their isolated ecosystem. I've done some light and SEM on samples but need to get inside to determine if the bacteria that break them down are carried internally or are picked up environmentally from the water. For sample preparation and sectioning I need to consider other contents like diatoms and stromatolite pieces and the high concentration of mineral solids in the samples. To start I plan to fix with glut/OsO4, dehydrate with acetone, infiltrate with propylene oxide and go into embed 812. After sections are imaged for non organic I planned on staining with lead citrate and comparing. Being new to TEM and not having a good idea of the precise osmolarity of the sample has left me puzzling over buffer selection (sodium cacodylate vs. phosphate), concentration, and times. We have an excellent EM technician here at NAU but it seems more my responsibility to make these determinations so any suggestions or references you could offer would be greatly appreciated. Thanks Pete Polsgrove Northern Arizona University pjp6-at-dana.nau.edu
we have just bought a Nikon CoolPix 990. It is an outstanding camera. One thing that we are disapointed it is a focusing problem when taking picture in Microscope. For any zoom setting and even in a manual mode, the micrograph borders always get out of focus, and in some situation with a Chromatic aberration. Does anyone have any idea how to eliminate this problem? The camera came with all necessary accessories for mounting in a microscope, e.g., c-mount, and MDC relay lens and remote cable.
Best regards,
Leonardo -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
The CoolPix 990 is indeed a nice camera. Unfortunately, it is not ideal for microscopy. It will work in this type of application but the results are mixed and the effort can be great.
If you zoom out to infinity, you should get best results. The key problem I have found is obtaining consistent exposure values. Preview with the puck looks good but the captured image is over exposed. Trying the best of five sometimes helps. But i have relegated the 990 to travel and snapshots. I don't think that it is ready from prime time microscopy. YMMV.
gary g.
http://photoweb.net
At 01:35 PM 3/2/01, you wrote:
} Dear All, } } we have just bought a Nikon CoolPix 990. It is an outstanding camera. One } thing that we are disapointed it is a focusing problem when taking picture } in Microscope. For any zoom setting and even in a manual mode, the } micrograph borders always get out of focus, and in some situation with a } Chromatic aberration. } Does anyone have any idea how to eliminate this problem? The camera came } with all necessary accessories for mounting in a microscope, e.g., } c-mount, and MDC relay lens and remote cable. } } } Best regards, } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
I am trying to distinguish between neurons and glia in cortical tissue that has been embedded in epoxy resin (LX112/NMA/DDSA/DMP-30). Does anyone know how to do this on semithin sections? Toluidine-Blue staining imparts some subtle differences in the appearance of chromatin in the nuclei of neurons vs. glia, but i am looking for a staining procedure that provides a more obvious color difference (without going all the way to immunohistochemistry).
It is my understanding also that KMnO4 will react with NMA (even if i disolve the resin out??), so those procedures are out, unless I can find a substitute for KMnO4.
let me make two observations and see if you agree:
1) The Coolpix (and any other camera like it) is originally designed for snapshots. That means, that the emphasis is on "nice" pictures under normal (outside or flash) conditions. This does not necessary mean, that one can take good scientific images. This may be the reason that you had problems with exposure time. The camera may take a weighted measurement to calculate exposure time, for example only from the middle. If you take a snapshot outside, the most important feature is likely to be found somewhere around the center of the image. So it's important to get that part right. That might not apply to photography on a microscope.
2) The quality of the picture is affected by the weakest link in the chain. My suspicion is, that the lens on the camera is far inferior to anything you put on a microscope. For example, we spend a lot of time (and money) to develop and make our own lenses for our TEM cameras. I don't see the point of spending thousands of Dollars on good microscope lenses and then have all that negated by an inferior lens on the camera.
One word to Nikon: I am not trying to knock this camera, it is probably a very good camera (I don't know it myself). My remarks are general and apply to other camera manufacturers (Kodak, Canon, Fuji, etc.). This was just a thread which mentioned the Nikon camera. Plus, all these ramblings are my personal opinion.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, March 02, 2001 10:20 PM To: Leonardo Lagoeiro Cc: MSA listserver
The CoolPix 990 is indeed a nice camera. Unfortunately, it is not ideal for microscopy. It will work in this type of application but the results are mixed and the effort can be great.
If you zoom out to infinity, you should get best results. The key problem I have found is obtaining consistent exposure values. Preview with the puck looks good but the captured image is over exposed. Trying the best of five sometimes helps. But i have relegated the 990 to travel and snapshots. I don't think that it is ready from prime time microscopy. YMMV.
gary g.
http://photoweb.net
At 01:35 PM 3/2/01, you wrote:
} Dear All, } } we have just bought a Nikon CoolPix 990. It is an outstanding camera. One } thing that we are disapointed it is a focusing problem when taking picture } in Microscope. For any zoom setting and even in a manual mode, the } micrograph borders always get out of focus, and in some situation with a } Chromatic aberration. } Does anyone have any idea how to eliminate this problem? The camera came } with all necessary accessories for mounting in a microscope, e.g., } c-mount, and MDC relay lens and remote cable. } } } Best regards, } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
Does anyone have a GaAs cathodoluminescence attachment in a SEM? This means a photomultiplier optimized to 8500A, an S1 photomultiplier. I would like to look at light emitted from a GaAs diode under power. Or does anyone know of a lab that does have GaAs cathodoluminescence as a SEM attachment.
I would like to thank those colleagues who kindly provided their advice and shared their experience with us regarding the problems of vibratome brain sectioning that I posted on the list server a while ago.
We now use double-edge feather blades that we purchased from Ted Pella, the quality of sections are much improved, also sometimes it seems helpful at low amplitude rather than high one.
Xinran Liu, M.D., Ph.D.
Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4. 214A Dallas, TX 75390-9111
Arron: Try using Araldrite 502 for your resin. It does not contain NMA. This should eliminate your concern with using KMnO4. The mixture I use is a 1-1 Araldrite 502 and DDSA, mix well then add your DMP-30 and mix again. Don't worry about the air bubbles.
Ron Austin (Research Associate) Dept of Pathology LSU Medical Ct. Shreveport, LA rla-at-mindspring.com
-----Original Message----- } From: "PostMaster {rla-at-mindspring.com} "-at-sparc5.microscopy.com [mailto:"PostMaster {rla-at-mindspring.com} "-at-sparc5.microscopy.com] Sent: Saturday, March 03, 2001 2:43 PM To: rla-at-mindspring.com
-----Original Message----- } From: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Saturday, March 03, 2001 2:41 PM To: Microscopy Society of America
Email: pjp6-at-dana.nau.edu Name: Pete Polsgrove School: Northern Arizona University
Question: I'm looking for an protocol for imaging fecal pellets. The pellets are from a hydrobiid snail found in highly mineralized warm desert springs and Posos in Mexico. I am interested primarily in the bacteria found on and in them but would also like to be able to inventory the entire contents of the pellets to help get a better picture of what part they play in their isolated ecosystem. I've done some light and SEM on samples but need to get inside to determine if the bacteria that break them down are carried internally or are picked up environmentally from the water. For sample preparation and sectioning I need to consider other contents like diatoms and stromatolite pieces and the high concentration of mineral solids in the samples. To start I plan to fix with glut/OsO4, dehydrate with acetone, infiltrate with propylene oxide and go into embed 812. After sections are imaged for non organic I planned on staining with lead citrate and comparing. Being new to TEM and not having a good idea of the precise osmolarity of the sample has left me puzzling over buffer selection (sodium cacodylate vs. phosphate), concentration, and times. We have an excellent EM technician here at NAU but it seems more my responsibility to make these determinations so any suggestions or references you could offer would be greatly appreciated. Thanks Pete Polsgrove
1) The camera is of course intended for general purpose photography. Snapshots, indoors and outdoors. It has a miniature flash which is good for about ten feet distance. For microscopy, of course the flash is turned off.
2)There are some macro zoom situations where the camera does a nice job. But for compound transmitted and reflected work, I have not found it to be satisfactory. Optem has made a high quality interface for the CoolPix which interfaces to Zeiss, Olympus and many other 'scopes. I have a common optical tube with C-mount to CoolPix for Axioskop and Olympus. I cannot fault this adapter system at all. But as you point out, the native CoolPix optics are not intended for discriminatory microscopy work. At about $700 for the Optem adapter, the CoolPix package with remote release and AC adapter is not cheap.
I recently took 765 pix with the CoolPix on a trip to Australia. It did a super job. I used two 128MB AVL CF modules. Some images were in highest JPEG resolution and some were in XGA fine. All print on an Epson Stylus Photo 2000P with outstanding results. And, of course, they go to the Web without any problem.
The Pixera Penguin 600CL is quite a different camera. It is highly suitable for microscopy, but also will do excellent macro work with an inexpensive C-mount zoom lens. Optem also makes the adapter for this camera to the Zeiss and Olympus microscopes. Same high quality and fine results. I have done multiple slices and stitched them together for a final TIFF file size of 110MB.
The Pixera is non-interpolated 5.8M pixels using their Diractor prism device. At 2776x2074 pixels, it generates a 17.5MB TIFF file. In 16-bit mode, the size is twice that. As I mentioned in a prior post, their software supports multiple capture averaging and integration. It also does variable sized spot metering and average metering. The CL version has Peltier cooling and exhibits exceptionally low thermal noise. Four frame averaging reduces noise even further.
The Pixera is 10-bits per color while the Nikon MX1200 is 8-bits per color. The word on the street is that the next release of the MX1200 will be 10-bits per color. Whether this is worth anything to a particular user is their own decision.
Being a professional Nikon shooter for many years, I am no longer an intransigent fan. In fact, I have dumped nearly all of my Nikon SLR equipment in favor of Contax. This of course has nothing to do with microscopy. But the point is that in my extended experience, Nikon is struggling to develop new products, which by specifications, leapfrog their competition--while in practice, they do not.
Try before buy is a good operative in this respect. I really don't care who makes what, as long as it is good stuff. Sorting through all the marketing hype is a chore. Sometimes even working with the equipment is a chore. But the actual results of using products is much more telling than sales pitches or product brochures.
gary g.
http://photoweb.net
At 06:54 AM 3/3/01, you wrote:
} Gary, } } let me make two observations and see if you agree: } } 1) The Coolpix (and any other camera like it) is originally designed for } snapshots. That means, that the emphasis is on "nice" pictures under normal } (outside or flash) conditions. This does not necessary mean, that one can } take good scientific images. This may be the reason that you had problems } with exposure time. The camera may take a weighted measurement to calculate } exposure time, for example only from the middle. If you take a snapshot } outside, the most important feature is likely to be found somewhere around } the center of the image. So it's important to get that part right. That } might not apply to photography on a microscope. } } 2) The quality of the picture is affected by the weakest link in the chain. } My suspicion is, that the lens on the camera is far inferior to anything you } put on a microscope. For example, we spend a lot of time (and money) to } develop and make our own lenses for our TEM cameras. I don't see the point } of spending thousands of Dollars on good microscope lenses and then have all } that negated by an inferior lens on the camera. } } One word to Nikon: I am not trying to knock this camera, it is probably a } very good camera (I don't know it myself). My remarks are general and apply } to other camera manufacturers (Kodak, Canon, Fuji, etc.). This was just a } thread which mentioned the Nikon camera. Plus, all these ramblings are my } personal opinion. } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Friday, March 02, 2001 10:20 PM } To: Leonardo Lagoeiro } Cc: MSA listserver } Subject: Re: Nikon CoolPix 990 } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } } Subject: } } } } SEM views of gold paint } } } } Content: } } } } Does anyone have any views of modern gold flake (real gold, not brass } } flake) used in making paintable substitutes for gold leaf? } } } } John Twilley } } Conservation Scientist }
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The candidate will be working in a diverse industrial R&D-environment with several links to university labs. He/she will be involved in a multidisciplinary team working on several nanostructured applications. Main focus of the work will be on microscopical and crystallographic (diffraction) techniques. In order to meet the technical requirements, the candidate should have a PhD in materials science, experience with TEM- and diffraction-techniques, and have a background in crystallography of inorganic materials. He/she should be a good teamplayer to function in a multi-disciplinary team, and be fluent in English and/or Dutch.
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For selection procedures, eligibility criteria and so on, please see homepage Marie Curie Industry Host Fellowships: http://www.cordis.lu/improving. Please note in particular that candidate fellows must be nationals of an EU Member or Associated State, or have resided in the EU for at least five years immdiately prior to their selection by Agfa.
I don't want to wake up a recent discussion (I was away in the last weeks), but I take advantage of this subject to askh a question about an other aspect of this subject. Sometimes you have so old material and maintenance services are laughing when they see what your are working with and/or on the other hand (with new materials also) it's faster and cheaper to do cleaning and (basic) maintenance yourself (and I like to do it myself !).
My question : What kind of grease/oil, where, on a LM ?
Manufacterer and maintenance services don't like to answer such a question. They say its complicate, there are much type of greases etc etc... So, does someone know something about that.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I was quite surprised to see the problems that are being discussed in relation to TEM focus, particularly as I was with Hitachi in the HU11 days, lets keep it simple and try to help?
When setting the image focus you should be aware of a number of problems that may complicate matters.
1. To try to focus at condenser cross over is absolutely the wrong thing to do! i) At crossover there is the poorest illumination coherence, this will lead to soft images, not out of focus but not optimised either. ii) At crossover there is a space charge effect (too many electrons in a small area and they repel each other) which may cause a false focus. (Just look at any image at crossover and then overfocus the condenser - you can see more detail!) iii) To focus at crossover and then change to a lower intensity is asking for trouble as the change in heating effect is likely to cause specimen drift and flexing which may change the contrast of material science specimens.
2. You MUST focus at the photographic magnification. Focus is the matching of the focal lengths of the objective and diffraction lenses. If you focus at a higher level and there is a diffraction lens change when dropping the magnification, you will find a matching objective correction will be required or your image will be out of focus.
3. What is best FOCUS, not always true focus? To a materials scientist it will almost certainly be true focus, no Fresnel fringes, as determined by the focus wobbler. They will set their intensity in advance as an intensity change may cause the material to flex and change in contrast. To the biological scientist it will be a degree of underfocus determined by the specimen's organelle density, its thickness, the kV, the objective aperture size and most important, the magnification; this is optimum under focus, the defocus where the (white) Fresnel fringes enhance the high contrast areas!
When taking a photograph the ideal conditions (as used in all the manufacturer's demo laboratories that I have worked with, Hitachi, JEOL and ISI) require the photograph to be taken at the same intensity level as when the focus is set. WHY? Well, once an intensity is set the operator may focus and during the focus procedure they should be concentrating sufficient to spot any image drift; it is most important to see the image under the conditions that you are going to photograph. So what will the excuses be?
1. "It is too dim to focus" - then i) increase the emission current, many people run at too low a current and make the task of optimising the instrument settings much more difficult ii) increase the kV which would increase the intensity too ii) re calibrate the photographic system to allow a focus intensity that is suitable for most operators
2. "The negatives are to dark" - i) the denser the negative the higher the contrast, so may be you could increase the kV, have a more stable specimen and an even brighter image?
3. Difficult one this - "the pathologists like a very bright image" - yes I know I guess it comes from looking down light microscopes all day? i) try 1 or 2 above - "yes but the pathologists like a high screen contrast" - yes I know but you should explain that no one publishes the screen, it it the photographic record that is the most important item(?) and we can get stacks of contrast from modern printing/publishing systems.
Regarding the reported overlap in currents between condenser and objective it is true, however the result is usually more of an image shift than a focal change in my observations, but yet another reason to focus and stigmate at the photographic intensity.
So to conclude I would
1. Always focus and stigmate at the photographic illumination level, if a biologist determine and plot out the optimum under focus for your material over your normal magnification range. 2. Always use the second condenser overfocus from crossover for high coherence and combine this with a 2 micron spot size for biology. 3. Set up the instrument's photographic procedure to suit as many operators as possible so that they too may focus at the photographic level. 4. Increase the emission current to give a higher brightness making point 2 far easier to use 5. Always use a higher kV (if 60, wow please go to 80, if 80 try 100) and work to obtain the best results on a photograph not trying to optimise for the screen. 6. To focus at low magnifications try removing the objective aperture and focus without a binocular, or a wobbler, looking for the very strong minimum contrast effect.
Well I hope this helps, its standard information on our courses, guess we do too much SEM in the States!
Regards
Steve Chapman Senior Consultant Protrain For consultancy and training by professionals World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
} } I have several questions for an AFM microscopist; } } 1. Can AFM really measure surface modulus?
Probably - but there are a number of factors which may have to be taken into account. These will include the nature of sample, any additional interactions between the sample and the scanning probe tip, the nature of the tip and SPM cantilever and so on.
} } 2. If yes, what is the mode of operation or measurement? } In terms of imaging modes (i.e. those where you are obtaining an image along with surface property information) there are two that may provide some surface modulus information.
i) Phase imaging (see for e.g.http://www.di.com/AppNotes/Phase/PhaseMain.html) is capable of providing a surface map based on the viscoelastic properties of the surface. Therefore the image will probably be a convolution between both the surface modulus and other surface properties (for example elasticity and whether the sample has any interactions with the tip). In this mode the probe is tapped into the surface and the lag between the voltage driving the tip oscillation and the actual tip oscillation provides this information. (It is particularly difficult, if not impossible to quantise the results obtained).
ii) Pulsed force should be able to deconvolute between sample stiffness and sample adhesion. http://www.thermomicro.com/tech/modes/pfm.htm should provide more information about this mode. However whether true values could be measured would again be a matter of some discussion.
Further mode of scanning probe microscopes are the spectroscopic ones in which the probe tip is move towards and away from the sample and the tip response measured. In AFM this mode is force-distance (see for e.g. http://www.thermomicro.com/tech/modes/fvsd.htm). This should again provide some information regarding the surface modulus. And with some tip calibration may provide something akin to a quantitative measurement.
With all of these mode the cantilever type will have an effect on the amount of information available. If the cantilever is less stiff than the surface of interest your measurement will generally be of the Young's modulus of the cantilever.
Nanoindenters may provide a more accurate measurement. http://freespace.virgin.net/micro.materials/ is an example. There are some companies that offer methods of converting commercial SPMs towards a nanoindenter.
} 3. Is it a quantitative measurement? Is it an absolute measurement } or a relative measurement?
My cynical opinion would be that most measurements available from a more qualitative than quantitative, though many would disagree. For example in this case, the area of tip-sample contact is unlikely to be able to be measured accurately but will have a major effect on your measurements, You may however be able to obtain some numbers that can be converted to an absolute measurement after careful consideration.
} } 4. Can the results be correlated to the (bulk) modulus measured by } rheometer?
This would, I presume, depend upon the material.
I hope this is of some use.
Giles
---------------------------------------------------------------------------- --------------------------- Dr. Giles Sanders Zeneca / SmithKline Beecham Centre for Analytical Sciences Chemistry Department Imperial College of Science, Technology and Medicine London SW7 2AY
I have had a request from a customer to allow remote access to our SEM in operation ( Hitachi S3500N ). The software they are proposing to use is "Timbuktoo" (?). They wish to set up three-way access across the internet along with a web-cam. I would appreciate advice from anyone who has tried this already. It would be useful to know what software has been used elsewhere and what problems have arisen. There may be better software available to carry out this function, but I have not approached this problem until now and am a bit in the dark. All advice, comments, etc. will be much appreciated.
Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
Dear Colleagues We have a CM200 (S)TEM equipped with EDAX IIs, 4pi digital imaging system and Gatan GIF. We are considering buying an integrated data acquision system for this microscope. ES Vision System from Emispec Systems, Inc. is under consideration . Comments from users of this ES Vision System would be highly appreciated. Of particular interest is operation of the Gatan Image Filter; would the autofilter functions still be available? Are there any other such systems out there? TIA Anita
To the person who is polymerizing the L.R. White at 60 degrees C:
Turn down the oven. If you want to do some sort of gold labeling on your sections try polymerizing the blocks at 45 degrees C for 2 to four days. Be sure to do this in some sort of BEEM capsule, as the stuff will not get hard if it is exposed to air. This type of plastic is not very stable in the electron beam, pick up your sections on carbon coated gold grids. Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
First off, I suppose that Nestor or the folks at ORNL will have much more to say about this since they have essentially developed and refined telemicroscopy. However, I have some experience with Timbuktu that I will relate.
We have been using Tibuktu for a number of years since I first heard of it several years ago at an MSA workshop on telemicroscopy. It is designed for remote observation or control of computers in general and has been borrowed for the microscopy application. There are several other products also available for the same work. Norton has PC anywhere. ATT has developed VNC which has been mentioned here before and is free.
Timbuktu (and the others) simply requires a connection to the Internet. Whatever happens on the screen is potentially available to the world once a client logs in with a password. This can slow down the host computer as it also has to transmit a copy of the screen.
I don't know about PC Anywhere, but Timbuktu is much faster on the client side than is WinVNC, even on a LAN. There can be a lot of data to transmit and it flat out takes a while to do so. Well-written code can do a lot to speed that up. You should also consider the connection between your PC and your client. If they have only a modem connection, refresh rates will be painfully slow.
I couldn't guarantee how well Timbuktu (or any other program) would work on your scope. I seem to recall that there were some windows whose contents were not available for Timbuktu to share. It may be that the SEM control programs were doing non-standard operations that circumvented the normal Windows system. Presently, I am able to view video windows whether they be from Oxford's AutoBeam, Quartz's PCI or Dazzle's video preview. But don't expect refresh rates measured in frames per second, at least not with a remote control/viewer program. Still, it could be a good alternative compared to more expensive solutions or to traveling to the site.
BTW, we also picked Timbuktu since it worked with either Macs or Windows machines.
I cannot tell you much about web cams. I know some systems are available that are stand-alone and practically plug and play. I don't know that I would want to burden my microscopy or EDS computer with the extra task. Others may have more to say.
There, that ought to be worth a couple of cents.
Warren
At 03:57 PM 3/5/2001 +0000, you wrote: } Hi, } } I have had a request from a customer to allow remote access to our SEM in } operation ( Hitachi S3500N ). The software they are proposing to use is } "Timbuktoo" (?). They wish to set up three-way access across the internet } along with a web-cam. } I would appreciate advice from anyone who has tried this already. It would } be useful to know what software has been used elsewhere and what problems } have arisen. There may be better software available to carry out this } function, but I have not approached this problem until now and am a bit in } the dark. } All advice, comments, etc. will be much appreciated. } } Thanks. } } Best wishes, } } Colin
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Timbuktu is a remote control software package made by Netopia.
http://www.netopia.com/software/
There is a free trial version which can be downloaded.
Whether it will work for you, I don't know. One of the key issues is real-time image transfer for mag and focusing. The interconnect between the SEM and the remote user is a major factor in response time. Slow analog modems are not going to be all that responsive. If running on a LAN or intranet, that would make a big difference.
A similar product is PC-Anywhere.
gary g.
At 07:57 AM 3/5/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You can do it very cheaply (i.e. free if you have Windows machines) using NetMeeting. Our LEO 1530 was easily set up to do this. You can do "over the shoulder" microscopy where the remote location observes. You need fast bandwidth for control and the instrument must be fully digitally controlled. We have tried it out with remote locations within our company and it works, but do not use it very much (i.e. never).
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
-----Original Message----- From: Colin Reid [mailto:creid-at-truxa1.tcd.ie] Sent: Monday, March 05, 2001 10:57 AM To: MSA.Microscopy.Com (E-mail) Subject: Remote Access to SEM
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html --------------------------------------------------------------- --------.
Hi,
I have had a request from a customer to allow remote access to our SEM in operation ( Hitachi S3500N ). The software they are proposing to use is "Timbuktoo" (?). They wish to set up three-way access across the internet along with a web-cam. I would appreciate advice from anyone who has tried this already. It would be useful to know what software has been used elsewhere and what problems have arisen. There may be better software available to carry out this function, but I have not approached this problem until now and am a bit in the dark. All advice, comments, etc. will be much appreciated.
Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
to make a guess, you should can give some more information. What impurity elements did you find/expect ? Can you make any suggestion about the size of any kind of inhomogeneity - conzentration, precipitation or what ever ?
S.Guder
} We're analysing Cu wire here for purposes better known to ourselves. We } are noticing, but not able to image interesting variations in the } concentration of impurities, which suggests to us, either that the } original material was inhomogeneous, or that drawing the wire has caused } some kind of exsolution process. Have any of you encountered smilar } things in metals? Refs? } Malc. } } -- } Dr MP Roberts Phone: [+27](0)46 603 8313 } Dept of Geology Fax: [+27](0)46 622 9715 } Rhodes University Cell: 083 4060 262 (usually off) } 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za } SOUTH AFRICA
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr.-Ing. Susanne Guder Technical University Munich Department of Materials in Mechanical Engineering D - 85747 Garching Phone: + 49-(0)89-289-15308/15338 Fax: + 49-(0)89-289-15301 Email: guder-at-wm.mw.tum.de ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I own a zeiss 63X water immersion lens. If the coverslip thickness is 0, does that mean that this is meant for direct viewing of specimens in water? I purchased it with that in mind (I work in marine environments, felt it would be convenient to observe microorganisms and microanimals directly). Using a coverslip doesn't seem to work well, if at all. For some specimens it is nice, but any focusing movement will push specimens out of the way. Nice for zooxanthellae in a small sea anemone, for example, and was pretty good for a thick broth of unicellular green algae.
I am trying to get a feel for this objective. WOuld appreciate any information possible.
Alan Davis adavis-at-saipan.com
On Fri, 13 Oct 2000 08:51:07 -0400 Gary Radice {gradice-at-richmond.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I've had some experience with water immersion lenses. Their main } advantage is higher numerical aperture than dry lenses at } equivalent } magnification, so better theoretical resolution. They don't offer } quite as good resolution as oil immersion lenses, but they tend to } have longer working distances, I believe, and don't have the } messy } clean-up of oil immersion lenses. So, yes, they do have some } distinct } advantages. } } However, as others have pointed out, you may already be able to } see } everything you need to see with your current lenses. And nothing } beats a test-drive. } -- } Gary P. Radice gradice-at-richmond.edu } Associate Professor of Biology 804 289 8107 (voice) } University of Richmond 804 289 8233 (FAX) } Richmond VA 23173 http://www.science.richmond.edu/~radice
-- adavis-at-saipan.com 1-670-235-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, NMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
Has anyone priced the XL30 high vac FESEM and the EFESEM or variations which use LaB6? What is the general price range for these, and which Hitachi models would closely compete?
A normal (upright) epifluorescence microscope excites fluorescence in a specimen by passing the excitation light to the top of the specimen via the objective lens, which is used to focus the light into an intense spot exactly at the point on the specimen which is to be viewed. Fluoresced light from the region of interest is collected by the objective and is viewed or photographed normally after filtration to remove any excitation light and unwanted fluorescence wavelengths. The excitation illumination is commonly provided by a high-pressure mercury vapour lamp, which conveniently provides very high intensity illumination in UV, blue and green wavelengths. Other high-intensity sources are now also used, including xenon lamps, lasers (e.g. in confocal microscopes, which are a highly-derived type of epifluorescence microscope) and even tungsten-halogen lamps. The separation of excitation and fluorescence wavelengths is usually accomplished using a dichroic beam splitter which reflects green, for example, and transmits orange/red. Further fine-tuning of the excitation wavelengths may be done using dichroic filters. Many microscope manufacturers provide filter systems as beam-splitter cube assemblies with all the filters and dichroic reflectors required for a particular fluorochrome installed in one pre-aligned package for easy exchange. The fluoresced light may be further filtered for viewing or photography by "barrier" filters either of dichroic type, or of coloured glass or sometimes gelatin (e.g. Kodak Wratten).
Other than that, an epifluorescence microscope can be a normal compound microscope, but because efficiency of illumination and collection of light is important, and is limited by the objective lens, fluorescence microscopes usually employ high quality objectives with very high numerical aperture. If UV is used, it may be necessary to select ojectives with very low UV absorption.
Hope this helps Chris Date sent: Mon, 5 Mar 2001 17:25:50 -0600 To: Microscopy-at-sparc5.microscopy.com } From: ileyozerlat-at-yahoo.com ()
The FEI XL30SFEG costs in the region of £250k in UK The closest Hitachi competitor is the S4700. The FEI is thermal field emission, the Hitachi is Cold cathode field emission. There is no Hitachi ESEM, but they do a variable pressure tungsten filament SEM.
Philips / FEI service in UK is excellent. I have no first hand knowledge of the Hitachi service operation, but believe it is also first rate. Hitachi quote much lower prices for their service contracts than FEI. They tell me that their engineers generally have little to do because the instruments are so reliable! I don't know whether this is a true reflection of the cost of ownership, however.
Date sent: Tue, 06 Mar 2001 02:03:30 -0800 To: MSA listserver {Microscopy-at-sparc5.microscopy.com} } From: Gary Gaugler {gary-at-gaugler.com}
Hello,
Our Siemens Elmisjop 102 is presently out of order. We are looking for several parts of a Siemens Elmiskop 102 which must be replaced.
Parts List C73000 - E3174 - C1 - 1
Part N°335 - Objective current control fine / superfine - cat n° C72315 - A29 - A4
Part N°333 - Objective current control coarse I level - cat n° C72315 - A31 - B1
Part N°334 - Objective current control medium II level - cat n° C72315 - A31 - B2
Part N°455 - P.C. board V9 "Objective controller" LR - cat n° C72302 - A41 - A2
Many thanks in advance
Best regards
Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis - Coordinateur Informatique de Division Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 mailto:philippe.drouillon-at-solvay.com
We have an Emispec Vision system on our CM200FEG TEM-STEM to acquire spectrum lines and spectrum images from our Oxford EDS and Gatan GIF. We tune the GIF using the autofilter functions, and setup the GIF with ImageFilterControl, but let ESVision read out the GIF multiscan camera during acquisition. All in all, we are quite pleased with the system. If you would like to discuss this offline, please contact me directly.
} Hi, } } I am trying to distinguish between neurons and glia in } cortical tissue that has been embedded in epoxy resin } (LX112/NMA/DDSA/DMP-30). Does anyone know how to do } this on semithin sections? Toluidine-Blue staining } imparts some subtle differences in the appearance of } chromatin in the nuclei of neurons vs. glia, but i am } looking for a staining procedure that provides a more } obvious color difference (without going all the way to } immunohistochemistry).
I think you are going to have to rely on morphology alone. Get a copy of Peters, Palay and Webster, "Fine Structure of the Nervous System". You should be able to extrapolate from the low mag EMs to LM. You could probably do an immuno for GFAP (most but not all astrocytes) but that still leaves oligos and microglia. I don't know of a good immuno for these cells on plastic sections. Plus, there will be smaller neurons to contend with. Sorry!
} It is my understanding also that KMnO4 will react with } NMA (even if i disolve the resin out??), so those } procedures are out, unless I can find a substitute for } KMnO4.
And the KMnO4 is for ??
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
There is an official open position now at Genentech (the one I alluded to in an earlier post, but HR is not too speedy over there) . The posting is below and can also be found on their website: http:// www.genentech.com {http://www.genentech.com/} .
For immediate consideration please send a c.v. to Peter Schow at pschow-at-gene.com {mailto:pschow-at-gene.com} . He is not on the list, so please email him directly. I have already forwarded previous resumes I received to him, so no need to resend those.
Position: Research Associate- Cytometry Lab Requisition #: 01-0003250
Description Research Associate position available in the Research Cytometry Lab. The lab support core facility, collaborative and basic research functions. Primary responsibilities include operation and maintenance of flow cytometers and/or imaging instrumentation and may cover all aspects of data acquisition, analysis, interpretation and presentation in the contexts of basic and applied research. Lab supports Coulter and BDIS flow cytometers, Leica (confocal) and Nikon (digital fluorescence) microscopes and both Mac and PC computers. The person will work in a dynamic and interactive environment requiring a strong teamwork orientation, good communication skills, flexibility and the ability to interact with a diversity of research personnel.
Requirements Position requires a BS/MS and extensive experience with flow and/or image cytometry. Good working knowledge of cell and molecular biology necessary. Industrial experience is desirable.
Holly Aaron Head, Berkeley Imaging Center University of California, Berkeley Dept. of Molecular and Cell Biology 447 LSA Berkeley, CA 94720-2751 hollya-at-socrates.berkeley.edu
Northwestern University is drafting a complaint against a vendor of microscopy products, AMT. Has anyone else had experience with lawsuits against vendors?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
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Nestor Your Friendly Neighborhood SysOp
=========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
I am considering this, but have read about issues of contrast perception and/or low light vision. It occurred to me that microscopists, particularly EM types, depend on their eyes in low-light situations more than any other profession I can think of, with the possible exception of cat burglars.
Has anyone out there had LASIK or similar procedure, and has it been positive or negative for your work? Did you notice reduced contrast sensitivity?
Thanks for any personal experiences; since it's not a direct microscopy query, please send replies to me and (if there's interest) I'll summarize for the list.
Rick Mott (myopic, -7 or so, with some astigmatism)
We have a ten year old cryo-stage and transfer device for sale.
It is a BioRad E7400 comprising: Pump valve controller Sputtering module Evaporation module Edwards E2M5 rotary pump x2 Valve block Transfer/coating unit with dewar Stage cooling unit with dewar Specimen insertion rod ie all you need to go cryo-SEMing.
The rod and flanges suit a JEOL 6400 SEM. The unit was functioning and in good condition prior to replacement. Any offers?
Eric Hines Microscopy Centre CSIRO Entomology Canberra.
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
It should be possible to do this much faster and without any sample preparation with X ray reflectometry.
See web site :
http://www-cxro.lbl.gov/optical_constants/
The is there the possibility to simulate some mesures.
Optical methodes could also be able to do this, by interferences in the TiO2 film.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
we are interested in purchasing a good software for desktop- analysis (on a PC) of X-ray EDS spectra, previously acquired on an analytical TEM and transferred to other computer.
In the book of Williams and Carter is mentioned only one such program, i.e. the DTSA, from NIST. Does anyone of you know a web address where I could get more informations about this software ? I would be delighted if there will be any possibility to check it before ordering, but where or how? Besides, does anyone know about a better program of this kind ? We would much better appreciate a free one, but we are ready to purchase even a commercial one.
Thank you for your attention.
Corneliu Sarbu, PhD Metallyrgy and Applied Materials Science Dept. Catholic University of Leuven Belgium
---------- } From: "CAROL.A.WALDER" {eencaw-at-elec-eng.leeds.ac.uk} To: mtlrmdb-at-leeds.ac.uk
Hi Listers,
I have a LKB Bromma 11800 Pyramitome whose belt has come loos and it needs to be replaced. I can't find the instruction manual or the slip of paper that my predecessor wrote the service person's name on.
Does anybody out there have a source for the instruction manual? I called Leica but they told me this machine was absolete when they acquired the LKB line.
I'll fix this thing myself if anyone can send me a copy of the manual (I'll glaly pay for the cost of copying and shipping).
If anyone knows of a repair person who still works on these ancient beasties, please let me know. It would be good to have someone who could give them tune-ups every now & then.
My screws are loose and I've got the hammer, blow torch sosme bubble gum and bailing wire....now all I need is the manual.
Pretty please??
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
An excellent write-up about the process is located at http://www.manufacturing.net/magazine/dn/archives/current/feature1.html
Cheers Peter Tarquinio peter-at-evex.com
Evex Analytical Microanalysis and Digital Imaging 857 State Road Princeton, NJ 08540 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com ----- Original Message ----- } From: "Rick Mott" {rickmott-at-alumni.princeton.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 06, 2001 11:20 PM
} In the book of Williams and Carter is mentioned only one such } program, i.e. the DTSA, from NIST. Does anyone of you know } a web address where I could get more informations about this } software ? I would be delighted if there will be any possibility to } check it before ordering, but where or how? } Besides, does anyone know about a better program of this kind ? } We would much better appreciate a free one, but we are ready to } purchase even a commercial one. } } Thank you for your attention. } } Corneliu Sarbu, PhD } Metallyrgy and Applied Materials Science Dept. } Catholic University of Leuven } Belgium
Corneliu,
DTSA is no longer sold as a Standard Reference Data product. NIST has abrogated its patent on DTSA so both the executable and source code are available from the following URL free of charge:
If you find it does not meet your needs, please drop me an email. Our group welcomes suggestions for new features or improvements.
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
Dear all, I send you the following message written by a serious student. I hope that someone can offer a possibility to her. Thanh you very much. All the best, Elena Belluso
My name is Elisa Fornero and I doing my Degree thesis, at Turin University - Italy- (Dept. of Mineralogical and Petrologic Sciences), on "INORGANIC FIBERS IN LUNG TISSUES: MINERALOGICAL CHARACTERIZATION, COMPARISION WITH NATURAL ASBESTOS AND CALCULATION OF DIFFERENT FIBROUS MINERALOGICAL SPECIES" (the samples of tissues derived from people not professionally exsposed in order to value the environmental pollution of fibrous minerals). During my thesis I extensively worked with SEM and EDS and I learned to prepare the samples from lungs tissues filtrated. I would like to have some international experience taking part, about for two months between April and June 2001, to a stage in a foreigner University. I would like found a laboratory where develop my knowledge in this field. If there is anyone interested to host me, I can send my curriculum vitae, Thank you Elisa Fornero
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
"I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain."
You can find information about DTSA at the NIST web site
http://www.cstl.nist.gov/div837/837.02/dtsa.html
There is only a Macintosh version of this software, so it may not be of use for you if you are looking for software for a PC. NIST has officially withdrawn DTSA as a Standard Reference Data product and has abrogated its patent. NIST is now releasing the software free from charge, however no longer officially supports the software except for internal development as an internal research tool for NIST. The NIST web site I directed you to has links to download the software application and the Pascal source code.
Tyrone Daulton
} Dear fellows microscopists,
} ... } we are interested in purchasing a good software for desktop- } analysis (on a PC) of X-ray EDS spectra, previously acquired } on an analytical TEM and transferred to other computer.
} In the book of Williams and Carter is mentioned only one such } program, i.e. the DTSA, from NIST. Does anyone of you know } a web address where I could get more informations about this } software ? ...
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Experienced Histotechnologist to work in Neuropathology Research Laboratory of Department of Pathology
The person we seek will be responsible for maintaining a neuropathology research laboratory in the Department of Pathology at SUNY Downstate Medical Center. Knowledge of immunohistochemistry, special stains, and histopathology is required. Previous experience in handling central nervous system tissue preferred. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in a laboratory with hands-on experience in tissue processing, histochemistry, immunohistochemistry, and operation of a light microscope. Experience with technical aspects of neuropathology and performing special stains including silver (Bielschowsky) and myelin stains.
Two years experience working on immunohistochemistry of CNS tissue specimens with knowledge of immunoreagents and experimental protocols.
BachelorUs degree in science desirable.
Laboratory skills including communications, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, operation of computers and office equipment.
Advanced computer skills (word processing and database management) essential.
Desirable Experience
Confocal microscopy desirable.
Salary commensurate with experience
Responsibilities
Purchasing supplies and equipment, budget reports, laboratory maintenance and brain banking.
Will operate all microscopic, photographic and computer equipment, and keep accurate records of all laboratory experiments and procedures. Light and fluorescent microscopy; tissue processing for paraffin embedding, sectioning and slide stainer for immunohistochemical procedures; computer imaging (PhotoShop); general photography; and library and web searches
Familiarity with computer software for keeping records, report preparation and table/figure construction using Microsoft Office software (Word, Excel, PowerPoint) as well as Endnote and Adobe PhotoShop.
Send resume to:
Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
------------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu ------------------------------------------------------------------------
Experienced Confocal Microscopy/Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for organization of a new facility that includes a confocal microscope and an electron microscope. The position includes overall management of the microscopy facility, user training, and user supervision. Requirements for the position include experience with light and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Experience with confocal and digital imaging techniques, microinjection, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization.
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
Excellent interpersonal and organizational skills are essential.
BachelorUs degree in science desirable.
Desirable Experience
Expertise in training in the operation of confocal microscope systems is a distinct advantage.
Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are critical. Significant facility with computers is desired.
Responsibilities
Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility.
Perform routine transmission EM, including tissue processing, ultramicrotomy, and examination; do preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing.
Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am reposting this for a colleague. If interested, please reply directly to
Ms. Linda McGuire lmcguire-at-downstate.edu
----------------------------------------------------------------------- Experienced Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for the overall operation of the EM laboratory in the Department of Pathology at SUNY Downstate Medical Center. This individual will oversee all aspects of specimen accession and processing, operation of the microscope, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
BachelorUs degree in science desirable.
Laboratory management skills including effective written/verbal communication skills to interact with a diverse group, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, and operation of computers and office equipment.
Desirable Experience
Previous experience in confocal microscopy highly desirable.
Previous experience in performing immunocytochemical staining and advanced computer skills usage (e.g. image analysis) is also desirable.
Salary commensurate with experience
Responsibilities
Maintain electron microscope in operating condition. Process clinical and research tissues for "thick" and "thin" sectioning. Darkroom management of photographic printing.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by: Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
{fontfamily} {param} Times {/param} {bigger} SCANNING ELECTRON MICROSCOPY (SEM) AND ENERGY DISPERSIVE
X-RAY SPECTROSCOPY (EDS)
WITH APPLICATIONS AT VARIOUS PRESSURES AND ATMOSPHERES
AN EDUCATIONAL SEMINAR-FRIDAY, APRIL 27, 2001
MOTOROLA GALVIN CENTER-SCHAUMBERG, IL
8:30 AM - 5:00 PM
{underline} Instructors {/underline} : Vern Robertson, JEOL-USA, and Nestor Zaluzec, Argonne National Laboratory.
{underline} Time {/underline} : Check-in at Galvin Center 8:30-9:00 am. Seminar 9:00-12:30 and 1:30-5:00.
{underline} Preregistration Required {/underline} : Deadline April 20, 2001; no walk-ins.
{underline} Cost {/underline} : Members of ASM-International and of the Midwest Microscopy and Microanalysis Society-$30; Non-members-$50; Students: $10. Registration includes lunch and refreshments at breaks. Check or money order to accompany preregistration form (below).
{underline} Exhibits {/underline} : Representatives from the following industrial co-sponsors will be present to discuss your applications and their products: EDAX, FEI, Hitachi, JEOL-USA, LEO, Noran, Oxford, and PGT.
{underline} Directions to Seminar Site {/underline} : Please see the other side.
{underline} Preregistration {/underline} : Please complete the following form. Mail it with your check or money order made out to "Chicago Regional Chapter of ASM-I" to Ms. Sheila Jungman, MSD 212, Argonne National Laboratory, Argonne, IL 60439 {bold} . {/bold} Deadline for receipt of the form is April 20, 2001. If you require a receipt for the preregistration cost, please so indicate below for pick-up at the Seminar Check-In desk. Questions? Phone 630-252-4157 or e-mail allen-at-aaem.amc.anl.gov.
{bold} SEM/EDS Educational Seminar-Friday, April 27, 2001
{/bold} Name (please print clearly)________________________________________________________
We are having problems with preparing brittle TEM sample of (single crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any suggestions beyond a kid-gloves approach?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Mary-Jacque, Several years ago at a used book library sale, I found a copy of Forensic Geology-Earth Sciences and Criminal Investigation by Raymond C. Murray and John C.F. Tedrow. It dates from 1975 and looks interesting. I hope to read more of it. Another title which came from a special session at the MAS-MSA meeting in 1985 is titled Electron Microscopy in Forensic, occupational and environmental health sciences by Samarendra Basu and James R. Millette. I still lament that the Microbeam Analysis Society dropped Sherlock Holmes' hat and pipe from their logo.
Lawry, I don't know about magnesium orthovanadate, but sapphire will work using the small angle cleavage technique. I have done cross sections of GaN on sapphire when I taught some students at Univ. of Illinois how to do it. John McCaffrey has done YBCO on cerium oxide on sapphire. If you are only interested in the bulk, that is even easier. If the magnesium orthovanadate is as brittle as you say, then SACT will probably work on that also. If you want, I can send you an image of the GaN/sapphire.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: L. D. Marks [mailto:ldm-at-risc4.numis.nwu.edu] } Sent: Wednesday, March 07, 2001 1:22 PM } To: Microscopy List } Subject: Brittle samples } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } We are having problems with preparing brittle TEM sample of (single } crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any } suggestions beyond a kid-gloves approach? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } }
A quick clarification before I get deluged with responses about how to do cross-sections; we are not trying to do this. All (?) we want is a "conventional" 3mm disc sample of sapphire or magnesium orthovanadate. No glue, no mounting on a Cu ring, no nothing.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
I think a good solution may be Tripod Polishing. We have an application note on preparing samples of sapphire with a GaN film which may be useful. Let me know if you would like a copy of the note and I can email it to you.
Best regards-
David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
} RE: Brittle samples } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are having problems with preparing brittle TEM sample of (single } crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any } suggestions beyond a kid-gloves approach? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } -------------------------------------------------------
Has anybody out there had any experiences with making your own transmitted electron detector for SEM? I'm interested in sharing ideas, references, etc. I've made one for myself and am fairly happy with its performance, but am wondering if there is more that I can do.
Thanks in advance,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have two questions for you immunolabeling experts.
First, a client has some precious and irreplaceable histological sections of human hippocampus that have been labeled with ABC - DAB. Will it be possible for me to de-paraffinize the sections, expose them to osmium vapor and re-embed them in resin on the slide and pop them off and resection them and see the DAB precipitate? Anyone have a protocol that has worked?
Second, these researchers plan to use Vibratome sections of mouse brain and immunolabel them for light microscopy using an ABC kit. I have done on-ultrathin-section colloidal gold immunolabeling for TEM. However, for this new project we would like to try pre-embedding labeling of 50-60 micrometer Vibratome sections, then embed and resection them for TEM. Knowing what works for light microscopy, we'd like to use streptavidin/colloidal gold or whatever for TEM visualization. My question is, of course, does anyone have a favorite protocol they would be willing to share? How do I keep the sections from curling? If we stick them on a glass slide to embed in resin and (hopefully) pop off later for resectioning, when and how do I get the to adhere?
Mahalo!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The URL I gave this morning for DTSA was correct, but not memorable. A few hours ago an alias was created to make it easier to navigate to the NIST Microanalysis Software web page. DTSA can now be accessed from:
http://www.nist.gov/dtsa
This page also contains links to Lispix, Dave Bright's excellent image processing application (now available for Win9x/WinNT/Win2k) and the NIST Monte Carlo codes. Lispix can be accessed directly at
http://www.nist.gov/lispix
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
Try epoxying a thin metal washer on one side of the sample, and dimple from the washer side.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: L. D. Marks Sent: Wednesday, March 7, 2001 10:21 AM To: Microscopy List Subject: Brittle samples
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
We are having problems with preparing brittle TEM sample of (single crystal) sapphire and magnesium orthovanadate via dimpling & PIPS. Any suggestions beyond a kid-gloves approach?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
The purpose of gluing a metal ring to brittle samples before dimpling is to support the sample. The sample is then waxed to the dimple grinding support stub and dimpled through the washer. We use this method routinely with brittle samples and not just cross sections. It's a good idea to carefullly clean the sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot washers usually work well for us, but we sometimes use other washer materials to avoid confusing Mo in the sample with Mo deposition artifacts.
Larry
---------- From: L. D. Marks Sent: Wednesday, March 7, 2001 11:38 AM To: Microscopy List Subject: Brittle samples - NOT CROSS-SECTION
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A quick clarification before I get deluged with responses about how to do cross-sections; we are not trying to do this. All (?) we want is a "conventional" 3mm disc sample of sapphire or magnesium orthovanadate. No glue, no mounting on a Cu ring, no nothing.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
the metallurgy or materials science group in your university should be able to tell you all about grain size measurements.
Best regards
Sam Purdy Technical Center National Steel Corp. Trenton Mi USA spurdy-at-nationalsteel.com
} ---------- } From: James Clarke } Sent: Wednesday, March 7, 2001 11:45 AM } To: NIH-IMAGE-at-LIST.NIH.GOV } Subject: countig grain size in archaeological iron samples } } IMAGERS! } Hope some one can point me in the right direction. I need to be } able to measure the grain size of metal samples which are of a } heterogenous nature. I am using a PC and Scion image seems to } have some of the things I need but some advice would be nice. } } --------------------------------- } James Clarke } J.Clarke1-at-bradford.ac.uk }
Fellow Microtomists: I am seeking information regarding 2 Sorvall MT2B ultramicrotomes that still are in use and in good working condition for both semi-thin sections and ultrathin sections. However, the faster speed of the return stroke of the cutting cycle no longer engages, even though the duration and position control belts are in tact for the slow-speed (cutting phase) of the cycle. Is this a relatively simple repair, and is there a company in the Cincinnati, Ohio, area that provides such repair service? [The serial numbers of these microtomes are 7701673 and 7800584. Do the first 2 digits reflect the year of manufacture (1977 and 1978 respectively)?] What would be a fair price for microtomes of this ventage? Thanks in advance for your assistance. EL Cardell
} } } Email: de-at-payload.com } Name: De McKeown } } Organization: Payload Systems } } Education: Graduate College } } Location: Cambridge, MA, USA } } Question: I need to determine the number of TV lines my microscope is } giving me. I have a test target but it only goes to 266 l/mm and I have } better resolution than that. I can't find a 'better' target and am stuck } trying to get an answer. How can I do this or where can I go for a new } target? } } I also have to find a contrast, but haven't got enough information yet to } ask a good question. } } Thank you!! }
There is a rash of virus mail out on the net again, a number of which are using "suggestive names" in the subject line. As a precaution some people trash messages which appear out of context.
While Humorous Titles are good for the soul (especially mine) and bring a smile especially when one gets the implied joke, let me remind you to at least add in the Subject Line with some standard notation so that individuals can more easily judge source of the mail. After all, we know that my Email filtering system is not perfect and suspect mail leaks through every so often.
Some Keywords include: TEM, SEM, EDS, .Confocal, LM, SamPrep, etc... check the FAQ site if you need idea's, but you can easily see what I mean.
For example: the posting on "I've got the screws loose" could have be preceeded or prepended by an appropriate Keyword like Microtome, even if the word Microtome was in () or abbreviated as uTome.
Unfortunately getting Mo (or anything else) on the sample destroys it as far as I am concerned. So, we have to rule out anything like this unless we can safely remove the metal with an acid (and not dissolve the oxide in the process).
On Wed, 7 Mar 2001, Thomas, Larry (PNNL) wrote:
} Laurie, } } The purpose of gluing a metal ring to brittle samples before dimpling is } to support the sample. The sample is then waxed to the dimple grinding support } stub and dimpled through the washer. We use this method routinely with brittle } samples and not just cross sections. It's a good idea to carefullly clean the } sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot } washers usually work well for us, but we sometimes use other washer materials to } avoid confusing Mo in the sample with Mo deposition artifacts. } } Larry } } ---------- } From: L. D. Marks } Sent: Wednesday, March 7, 2001 11:38 AM } To: Microscopy List } Subject: Brittle samples - NOT CROSS-SECTION } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A quick clarification before I get deluged with responses about } how to do cross-sections; we are not trying to do this. All (?) } we want is a "conventional" 3mm disc sample of sapphire or } magnesium orthovanadate. No glue, no mounting on a Cu ring, } no nothing. } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html } } } }
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
A researcher has asked me to localize gluconase (a protein) in plant roots. They provided me with antibody (sera) and preimmune, which they have used in westerns. The problem is the preimmune is labelling the cell wall; the labeling is quite impressive and very specific. Now they tell me (alas I didn't ask before) that yes they have background on the westerns, but the preimmune never labels the 32 Kd protein they are interested in. What is the best approach to localize this 32 Kd protein? Should we cross absorb everything else out, or would it be better to affinity purify for the desired antibody. And why does this rabbit have such good antibodies to cell wall?
Thank you, Michelle
Michelle Peiffer ************************************************************* Electron Microscope Facility for the Life Sciences Penn State University Biotechnology Institute 001 South Frear Lab University Park PA 16802
The key to thinning brittle hard samples is to start with a doubly polished thin section 30 micrometers thick. Cut out 3 mm diameter discs with an ultrasonic probe. Glue these to a glass slide with crystalbond, a thermal plastic cement that dissolves in acetone. Mechanically thin and polish to less than 10 micrometers or thinner. Ion thin until satisfied.
Start thinning a thin sample. Don't break it. One good sample is all you need.
I have made many ion thinned samples of brittle materials, mainly shocked minerals, for over thirty years. Also this method is not for those in a hurry.
Gordon Nord USGS Emeritus Scientist
"L. D. Marks" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Unfortunately getting Mo (or anything else) on the sample destroys it } as far as I am concerned. So, we have to rule out anything like this } unless we can safely remove the metal with an acid (and not dissolve } the oxide in the process). } } On Wed, 7 Mar 2001, Thomas, Larry (PNNL) wrote: } } } Laurie, } } } } The purpose of gluing a metal ring to brittle samples before dimpling is } } to support the sample. The sample is then waxed to the dimple grinding support } } stub and dimpled through the washer. We use this method routinely with brittle } } samples and not just cross sections. It's a good idea to carefullly clean the } } sample and washer so the epoxy bonds well. One- or two-mil Mo hole or slot } } washers usually work well for us, but we sometimes use other washer materials to } } avoid confusing Mo in the sample with Mo deposition artifacts. } } } } Larry } } } } ---------- } } From: L. D. Marks } } Sent: Wednesday, March 7, 2001 11:38 AM } } To: Microscopy List } } Subject: Brittle samples - NOT CROSS-SECTION } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } A quick clarification before I get deluged with responses about } } how to do cross-sections; we are not trying to do this. All (?) } } we want is a "conventional" 3mm disc sample of sapphire or } } magnesium orthovanadate. No glue, no mounting on a Cu ring, } } no nothing. } } } } ------------------------------------------------------- } } Laurence Marks } } Department of Materials Science and Engineering } } Northwestern University } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } } ------------------------------------------------------- } } } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } } http://xraysweb.lbl.gov/esg/phasing/index.html } } } } } } } } } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu } ------------------------------------------------------- } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
Nord Consultants 20594 Cornstalk Terrace Ashburn VA 20147
Below is the result of your feedback form. It was submitted by (linda_knoll-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 7, 2001 at 18:43:01 ---------------------------------------------------------------------------
Email: linda_knoll-at-msn.com Name: Linda Knoll
Organization: St.Joseph'sAcademy
Education: Graduate College
Location: St. Louis, MO USA
Question: How do the cells of humans and apes differ when looked at under a microscope?
Below is the result of your feedback form. It was submitted by (georgas1-at-home.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, March 7, 2001 at 20:29:09 ---------------------------------------------------------------------------
Email: georgas1-at-home.com Name: Adam Georgas
Organization: UMS-Wright Preparatory School
Education: 9-12th Grade High School
Location: Mobile, Alabama, USA
Question: Dear Microscopist: I am a 10th grade student working on my science fair project. My project deals with a computer program I wrote using fractal geometry to diagnose chronic lymphocytic leukemia. I obtained 25 pictures of normal cells taken from a Phillips electron microscope and 25 pictures of leukemic cells taken with the same microscope. I obtained these photographs from my mentor, a physician/professor at the University of South Alabama. My concern with my project is that the 25 healthy cells came from only one patient, and the 25 leukemic cells came from another patient. So, I have 50 cell pictures, 25 healthy, 25 leukemic, from only 2 patients. All of the research I have read use 25 different patients with only one cell from each patient. I am wondering if my study is valid? I posed this question to a local pathologist. She told me that since the pictures were taken from an electorn microscope, it was actually better to have more cells from just 2 patients. Is this true? If it is true, what is the rationale
Here is some parameters of 2000FX with AHP2OL polepiece: --------------------------------------------- Point resolution 0.31nm Lattice resolution 0.14nm OL focal length 4.1mm Cs 3.4mm Cc 3.1mm ---------------------------------------------
As to the question of beam semi-convergence angle, Prof. O'Keef has given a perfect note. I learnt a lot there as well.
Yours, Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
Fax: +49-6131-3923768 Tel: +49-6131-3923148(O) E-mail: syli-at-mail.uni-mainz.de; syli16-at-hotmail.com URL: http://syli.homepage.com/ (This URL contains my resume, SCI journal informations, JobList, TEMAlert, as well as other useful informations related to Transmission Electron Microscopy) ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Paula: Get in touch with Helmut Patzig at MOC, Spring Valley, NY. His email is Mocleica-at-aol.com. If anybody has the know-how and parts, I am pretty sure he does. Sorry I don't know of anyone closer to DC, but I have dealt with Helmut over the years with obsolete Reichert and LKB equipment.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Wed, 07 Mar 2001 08:31:14 -0500, Paula Sicurello wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi Listers, | | I have a LKB Bromma 11800 Pyramitome whose belt has come loos and it needs to be replaced. I can't find the instruction manual or the slip of paper that my predecessor wrote the service person's name on. | | Does anybody out there have a source for the instruction manual? I called Leica but they told me this machine was absolete when they acquired the LKB line. | | I'll fix this thing myself if anyone can send me a copy of the manual (I'll glaly pay for the cost of copying and shipping). | | If anyone knows of a repair person who still works on these ancient beasties, please let me know. It would be good to have someone who could give them tune-ups every now & then. | | My screws are loose and I've got the hammer, blow torch sosme bubble gum and bailing wire....now all I need is the manual. | | | Pretty please?? | | Paula :-) | | Paula Sicurello | George Washington Univ. Medical Center | Dept. of Pathology, Ross Hall rm 505 | Electron Microscope Lab | 2300 Eye St. | Washington, DC 20037 | 202-994-2930 phone | 202-994-2518 fax | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
The answer depends on the details of the "TV" system and microscope used. At this point I don't even know if it is an optical or electron microscope, etc... It should be noted that actual microscope resolution and the displayed resolution via TV have little in common.
NTSC (US) TV is 525 lines vertical (total, and the equevelant of 280 to over 640 horizontal (bandwidth dependent). Note that not all 525 lines are used for the image.
PAL, a standard used in many other areas of the world is slightly higher vertical resolution, but not much.
There are other "non-brodcast standard" resolutions used by custom instrumentation.
Woody
} -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } } } } } } Email: de-at-payload.com } } Name: De McKeown } } } } Organization: Payload Systems } } } } Education: Graduate College } } } } Location: Cambridge, MA, USA } } } } Question: I need to determine the number of TV lines my } microscope is } } giving me. I have a test target but it only goes to 266 } l/mm and I have } } better resolution than that. I can't find a 'better' } target and am stuck } } trying to get an answer. How can I do this or where can I } go for a new } } target? } } } } I also have to find a contrast, but haven't got enough } information yet to } } ask a good question. } } } } Thank you!! } } } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tina, Check the following reference for your answer to your second question:
Liposits, Zs., D. Sherman, C. Phelix and W.K. Paull. (1986). A combined light and electron microscopic immunocytochemical method for the simultaneous localization of multiple tissue antigens. Histochemistry 85:95-106.
We very successfully used vibraotomed sections of brain to do light and EM ICC. Becuase of the penetration problems for the gold conjugates available at that time, we used PAP-DAB for visualization on the light level followed by silver intensification of the material for EM visualization. The study involved serial sectioning of the material to locate synapses so flat embedding of the sections was manditory. The method is fully discribed in the paper but feel free to contact me if you have questions.
I would also try using the ultra small gold colloids presently on the market followed by silver intensification as an alternative to the PAP-DAB technique.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
I have two questions for you immunolabeling experts.
First, a client has some precious and irreplaceable histological sections of human hippocampus that have been labeled with ABC - DAB. Will it be possible for me to de-paraffinize the sections, expose them to osmium vapor and re-embed them in resin on the slide and pop them off and resection them and see the DAB precipitate? Anyone have a protocol that has worked?
Second, these researchers plan to use Vibratome sections of mouse brain and immunolabel them for light microscopy using an ABC kit. I have done on-ultrathin-section colloidal gold immunolabeling for TEM. However, for this new project we would like to try pre-embedding labeling of 50-60 micrometer Vibratome sections, then embed and resection them for TEM. Knowing what works for light microscopy, we'd like to use streptavidin/colloidal gold or whatever for TEM visualization. My question is, of course, does anyone have a favorite protocol they would be willing to share? How do I keep the sections from curling? If we stick them on a glass slide to embed in resin and (hopefully) pop off later for resectioning, when and how do I get the to adhere?
Mahalo!
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Michelle, The answer to the last question is easy. Remember Bugs Bunny munching his carrot? Rabbits eat plants so it is not too surprising that a preimmune serum recognizes a plant antigen, if not several.
What we have done in situations like this is to incubate the serum with the protein of interest (in this case, the 32kD glucanase) and then stain with that. This removes the specific glucanse staining. It is a kind of guilt by subtraction. It is not as nice as doing affinity purification, but it is a lot faster, and conserves the serum.
Hope this helps, Tobias Baskin
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listservers, we have been recently working on structural and compositional characterization of InGaAs/GaAs quantum dots grown by MOVPE, achieving interesting results. We have tried to use CERIUS2 to obtain simulations of cross sectional HREM images of these dots, but we experienced several problems. Is there anyone who has spent time on similar matters. We would like to discuss and compare our experiences, to understand if our problems are due to a bad construction of the supercell or to limitations of the software. Thanks Massimo
Thanks for all the responses to my original query. At the suggestion of Bart Cannon, you can check out my detector design at:
http://www.mta.ca/~jehrman/ted.htm
I perhaps should have clarified originally that this is a very simple design, and does not require any additional electronics, light pipes, PMT, etc. My machinist made it for me in an afternoon. I took inspiration from a single sentence in Goldstein, et. al (SEM and X-ray Microanalysis, 2nd edition, p. 267):
"A simple, inexpensive detector can be made from a high-atomic-number scattering surface placed below the specimen and tilted so that the transmitted electrons are scattered toward the conventional E-T detector in the specimen chamber"
} From my experience with one commercially available "real" TED a number of years ago, the homemade detector seems to perform as well (or better)
than the real thing.
No reference was given in the text, so I'm rather curious who came up with the idea originally. Are there any co-authors of the text lurking nearby who could shed any light?
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Question: Dear Microscopist: I am a 10th grade student working on my science fair project. My project deals with a computer program I wrote using fractal geometry to diagnose chronic lymphocytic leukemia. I obtained 25 pictures of normal cells taken from a Phillips electron microscope and 25 pictures of leukemic cells taken with the same microscope. I obtained these photographs from my mentor, a physician/professor at the University of South Alabama. My concern with my project is that the 25 healthy cells came from only one patient, and the 25 leukemic cells came from another patient. So, I have 50 cell pictures, 25 healthy, 25 leukemic, from only 2 patients. All of the research I have read use 25 different patients with only one cell from each patient. I am wondering if my study is valid? I posed this question to a local pathologist. She told me that since the pictures were taken from an electorn microscope, it was actually better to have more cells from just 2 patients. Is this true? If it is true, what is the rationale
Dear Adam, First, there is nothing about data from an electron microscope that makes sample selection any different from that used with other forms of data. The idea is that the population in your sample--in this case the normal and leukemic cells--should be representative of the larger population--all human normal lymphocytes and all human leukemic lymphocytes. If all normal lymphocytes in any one person have identical morphology, then any one cell will represent the population; if lymphocytes vary within an individual, then you would need a selection having the same distribution of types as exist in the whole person. This is usually obtained by selecting a small sample volume "at random". You might imagine how this could go wrong; e.g., if certain types were not present in arterial blood to the same extent as in venous blood. The next consideration is whether either normal lymphocytes or leukemic lymphocytes differ from person to person. I can see no reason that this would not be the case--especially if different forms of leukemia result from different transformation processes; e.g., different mutations or different extents of gene expression. In order to be sure that your sample is relevant, you would need to know whether there were different types of normal and leukemic cells both within and among individuals. This is an area where I am completely ignorant, but there is probably an extensive literature on this. There could also be differences arising from the preparation of the cells for EM, such as variation in fixation or staining, which can give apparent differences from nomonally identical cells. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi, I just started to bring up an ISI-SR50A. Followed the procedure to replace the filament, saturate beam current and aligment correctly but I could not get a decent image on the CRT even at low mag, 200x. Is there a way to check the detector? I varied the applied voltage from 5KV to 20KV and WD from 7mm to 40mm without any success. Any suggestions are welcome. -Vu.
__________________________________________________ Do You Yahoo!? Get email at your own domain with Yahoo! Mail. http://personal.mail.yahoo.com/
Here are two questions that I am pondering at the moment for thin film analysis in the TEM.
If you take the integrated intensities of the center portions of the peaks (zero crossing to zero crossing) of a top hat filtered spectrum to use for quantification, will you get the same results as you would if you do a background subtraction and take the integrated intensities from the peaks themselves?
For the integrated intensities of a top hat filtered spectrum, is it more appropriate to include the two negative lobes of the filtered peak added to the center portion?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
There are two freeware software packages that can accomplish this effort.
An IBM inspired software called OpenDX (for Unix), which is quite advanced. Start here (I have just installed this software but not worked with it yet):
http://www.opendx.org/
and through scion image, which is a PC version of NIH image. Start at the link below
http://rsb.info.nih.gov/nih-image/
You must install the ImageJ plugin, which you can obtain from:
http://www.isi.uu.nl/people/michael/vr.htm
This site also has other links for related information.
I am pursuing similar work with FIB/ Auger on particles and I would love to know about your progress. I believe this area has interesting and useful applications and I have found only very limited publications in this area. I would appreciate any additional information you might have.
Good Luck !
Regards, Ed
************************************************************ Edward Principe, Ph.D. Member of Technical Staff Defect & Thin Film Characterization Laboratory Applied Materials 408-986-3882
"Richard M Langford" {richard.langford-at-materials.oxford.ac.uk} on 03/08/2001 01:51:45 AM
To: microscopy-at-sparc5.microscopy.com cc: (bcc: Edward Principe/APPLIED MATERIALS)
Dear All
Can anyone suggest suitable software for 3-D reconstruction of grains and cracks from a sequential set of focused ion beam cross-sections.
Regards
Richard -------------------------------------------------------------- Richard M Langford
Department of Materials, University of Oxford Parks Road, Oxford, OX1 3PH, UK
If you only need to do reconstruction, download our free VoxBlast Light software at www.vaytek.com. Go to VoxBlast 3D software, then VoxBlast Light 3.0.
If you need any help or want more functionality, feel free to contact us directly.
Best regards,
Steve Niemela Sales sniemela-at-vaytek.com Ph: 641-472-2227 Fax: 641-472-8131
VayTek, Inc. 305 West Lowe Avenue Fairfield IA 52556 www.vaytek.com ----- Original Message ----- } From: "Richard M Langford" {richard.langford-at-materials.oxford.ac.uk} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 08, 2001 3:51 AM
A colleague is looking for a published reference that describes cholesterol crystals using transmission electron microscopy. We have searched the web but found only light microscopy of the crystals. Any help would be appreciated.
Thank you,
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
Facility Scientist (Academic Assistant III) Flow Cytometry and Confocal Imaging Facility, Biotechnology Center
The Biotechnology Center and the Department of Molecular and Cell Biology at the University of Connecticut invite applications for the position of Facility Scientist for the Flow Cytometry and Confocal Imaging Facility. This Facility houses a Becton Dickson FACSCalibur Flow Cytometer/cell sorter, a Leica SP2 laser scanning confocal microscope and several other microscope and image processing workstations. The facility scientist will be responsible for working with University faculty and students to develop research projects that utilize these instruments and related technologies. The facility scientist will also organize periodic training sessions and workshops to familiarize new users with the instruments. This individual will also be encouraged to develop collaborative research projects with University faculty and with local biotechnology businesses. We seek an individual who is excited by the challenges of an interdisciplinary research group and who enjoys interacting with people in an active scientific environment. The preferred candidate will have a Ph.D. in the biological sciences and familiarity with one or both of the core technologies. This is a full-time, annually renewable position. Salary will be commensurate with experience and education. Submit curriculum vitae, a statement of experience and interests, and the names, addresses, telephone numbers and e-mail addresses of at least three references to: Biotechnology Center, Confocal Search Committee, University of Connecticut, 184 Auditorium Rd., Unit 3149, Storrs, CT 06269-3149. Applications may also be faxed to (860) 486-5005 or sent electronically to biotctr1-at-uconnvm.uconn.edu. Screening of applications will begin immediately and continue until the position is filled.
We encourage applications from under-represented groups including minorities, women and people with disabilities. (Search #01A372) --
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
The following posting is for a colleague who is not on the listserver.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
We recently experienced a fire using a 5% perchloric acid - methanol electrolyte in a commercial jet electropolisher, and request information on similar experiences.
During operation of the electropolishing unit at room temperature and moderate pump speed, when turning up the voltage towards 30 V a loud pop was heard and the reservoir was found to be in flames. The fire was quickly extinguished using a dry fire extinguisher, but the reservoir and cable shielding melted.
We would appreciate any input on similar experiences using electropolishers with this or similar electrolytes. Any explanations for this behavior that you can provide would also be appreciated.
David S. Gelles Structural Materials Research Pacific Northwest National Laboratory Richland, WA 99353 Tel: (509) 376-3141 Fax: (509) 376-0418 E-mail: ds_gelles-at-pnl.gov
There are two ways that the top-hat filter has been used for quantifying EDS spectra:
(1) One can simply apply the filter to the spectrum as you suggest -- what results looks much like the second derivative of the original spectrum and each peak is represented by a central positive lobe with a negative lobe on either side. The slowly varying (continuum) components of the spectrum are suppressed. With care, one can relate the area of the positive lobe to the area of the original peak, however, there are some issues which make this simple concept a less than satisfactory quantitative method: (1) the area of the positive lobe is proportional to the area of the original peak, but the proportionality is a function of both peak area and filter shape. Thus the proportionality varies with the peak energy since the peak width varies with energy. (2) overlapping and adjacent peaks will interfere with each other, and this interference is actually increased by the broadening effect of the filter. Oddly enough, I have seen a number of familiar reference works which seem to give me credit for inventing the "top hat filter" used in this way -- I didn't. I saw it used this way for nuclear gamma ray spectra in the early '70s and after trying it myself, realized that it really wasn't satisfactory. But this experience did lead to the "filter-fit" method which follows.
(2) The"right" way for using the top-hat filter for continuum suppression is to use it in conjunction with linear-least squares fitting. One simply filters the unknown and each of the reference spectra to be fitted to it and then applies a conventional linear fit between them. When one fits the filtered references to the filtered unknown, the fact that the spectra have been distorted by the filter is immaterial (since the filter operator is linear), the overlaps are accurately unfolded, and the continuum basically drops out of the problem. Properly implemented, this is a very good quantitiative technique and has been employed productively for almost 30 years. If interested, the technique is published in NBS 604 (P273) or interested parties could contact me for references to several other papers describing it.
Fred Schamber
"Walck, Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Here are two questions that I am pondering at the moment for thin film analysis in the TEM. } } If you take the integrated intensities of the center portions of the peaks (zero crossing to zero crossing) of a top hat filtered spectrum to use for quantification, will you get the same results as you would if you do a background subtraction and take the integrated intensities from the peaks themselves? } } For the integrated intensities of a top hat filtered spectrum, is it more appropriate to include the two negative lobes of the filtered peak added to the center portion? } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
I've been gold immunolabelling whole retina pieces - 300um thick sheets - with great success (LM and EM) for years. I react the tissue in small vials, floating around loose. Briefly, you need a fixative like PLP or paraformaldehyde - any glut in the fix and antibody access to the tissue is decreased and you just get labelling on the cut edges. Use saponin in all solutions for penetration enhancement. Use an Fab2 anti Ig conjugated to 1 nm gold. Silver enhance then gold tone. Pop tissue in a silicon mould for embedding. If this sounds useful I can send you the full method.
As for rescuing paraffin sections, I did some a long time ago. From memory the resulting tissue looked like squashed newspaper - a mushy black mess with a hint of structure. I can't imagine even being able to distinguish the DAB from the rest, let alone see what was labelling. However, maybe methods have improved.
Cheers,
Diana --
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
In my opinion what happened was that a spark between the cathode and specimen, which often occurs at those voltages during electropolishing, ignited the electrolyte. I have used other electrolytes at voltages greater than 30V and I have also head a crackling noise which indicated to me that was sparking. Fortunately the electrolyte was not as ignitable as a 5% perchloric acid - methanol electrolyte. Nevertheless, I kept the voltage below that just in case for the electrolyte was still based on organic solvents.
You are going to be bombarded by horror stories and cautions about electrolytes containing perchloric acid. And they are right. I will not say that I have used electrolytes containing up to 30 vol.% in ethanol without problems because that would be tempting fate and I have enough Sicilian blood in me to make me a bit superstitious.
I have some recommendations.
1. As far as I know, perchloric acid electrolytes only require a voltage of 15V max. It is rare that I have used voltages in excess of 25V for any electrolyte and that was with an electrolyte of methanol, Butoxy-ethanol, magnesium perchlorate and lithium chloride (LiCl). This was the electrolyte where above 30V I was getting sparking. I have used water based electrolytes above 30V sometimes, but ignition is not an issue there.
2. Whenever making up or using an electrolyte containing perchloric acid, cool it down. When making it up, this prevents ignition due to heat of dissolution of the concentrate acid as it mixes. This may be paranoia. Cooling during electropolishing is necessary because it also reduces the risk if ignition and it improves the electropolishing process. I have found that most organic based electrolytes, especially those based on ethanol and methanol require cooling to {20°C in order to achieve the correct electropolishing conditions. So the necessity of cooling is two fold.
I hope this is useful.
"Thomas, Larry (PNNL)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The following posting is for a colleague who is not on the listserver. } } Larry Thomas } Pacific Northwest National Laboratory } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } We recently experienced a fire using a 5% perchloric acid - methanol electrolyte } in a commercial jet electropolisher, and request information on similar } experiences. } } During operation of the electropolishing unit at room temperature and moderate } pump speed, when turning up the voltage towards 30 V a loud pop was heard and } the reservoir was found to be in flames. The fire was quickly extinguished } using a dry fire extinguisher, but the reservoir and cable shielding melted. } } We would appreciate any input on similar experiences using electropolishers with } this or similar electrolytes. Any explanations for this behavior that you can } provide would also be appreciated. } } David S. Gelles } Structural Materials Research } Pacific Northwest National Laboratory } Richland, WA 99353 } Tel: (509) 376-3141 } Fax: (509) 376-0418 } E-mail: ds_gelles-at-pnl.gov
-- Dr. Steven Celotto Department of Engineering Materials Science & Engineering University of Liverpool Brownlow Hill Liverpool L69 3BX UNITED KINGDOM
I don´t know about the ISI50A but, assuming that it has the typical Everhard-Thornley design, isn´t it just possible that the bias voltage is inverted in your detector?. If the bias voltage is set negative, it´ll be rejecting all the secondary electrons, and you would only get a generally noisy image composed of the primary electrons backscattered in the very direction of the detector´s position. Quite nice for topographic information about flat specimens, but far worse quality than in "normal" SE images. Pardon me if I raised a too obvious explanation, but it occurred to me sometimes to get puzzled with a bad functioning of my SE detector just because another user of the microscope had been playing with the knobs.
Cheers
Diego
Diego Alvarez Instituto Nacional del Carbón INCAR-CSIC Spain
Thanks to all who responded to may plea for help in finding either a manual or a service person for the wee beastie.
Slowly but surely I will tighten the screws.....
Thanks again!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Full details of Trends in Nanotechnology (TNT) 2001, Segovia, Spain, September 2001, are now available. Based on the same format as last year, the conference aims to bring together researchers from all disciplines relating to nanotechnology, in a setting with ample time for interaction. A new feature this year will be the availability of grants to enable graduate students to attend and present posters.
Full details can be found at www.cmp-cientifica.com/tnt2001
Regards
Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
The NIST's web link to DTSA can be found at: http://www.cstl.nist.gov/div837/837.02/MicroscopySoftware.html
This program is free and available for the Macintosh only.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 100 Bureau Drive, Stop 8371 Gaithersburg, MD 20899-8371 http://www.nist.gov/cstl/div837/837.02/
TO be more specific, any advice on using formvar coated slot grids for serial sections. They seem to crinkle, or get blobby, right on the vessel I want to look at.
Ellen: Another question: are you using bare formvar or have you coated with carbon? Bare formvar will pucker, but the addition of a little carbon does wonders for stabilization and strength of the films. My experience is using 0.5% formvar, stabilized with about 10nm of evaporated carbon. These films aren't too thick, allowing good image resolution at 80kV. BTW: if the formvar films are puckered, they will show it prior to picking up sections. (Another vagrant neuron just fired.) One way to deal with the puckered formvar is to use chloroform or carbon tet. Simply pass the grid over the mouth of an open bottle, or, as a last resort, put the grid into the neck of the bottle (NOT into the fluid--just the vapors). Please be careful with this. (Exposure to these chemicals.) I may have the beginnings of interstitial pulmonary fibrosis--a not uncommon result of exposure to chemicals.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Sun, 11 Mar 2001 14:31:50 -0500, Ellen S. Morgan wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | TO be more specific, any advice on using formvar coated slot grids for | serial sections. They seem to crinkle, or get blobby, right on the vessel | I want to look at. |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Does anyone know of any buildings/laboratories recently constructed that house instruments (not necessarily EM's) requiring very low ( {0.5 mG) 60 Hz stray magnetic fields? I am interested in contacting someone (scientist, building manager, architect, etc.) who might know the details of how that building was constructed.
Thank you for your time and help,
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
At 5:24 PM +0100 3/9/01, Edmund Gierlik wrote: } Email: gierlik-at-delta.sggw.waw.pl } Name: Edmund Gierlik } Organization: Electron Microscopy Lab. } Education: professor } Location: Warsaw, Poland } } Question: Can anybody show the source of nice presentation of ESEM } application in a phase transition investigation (water - ice, liquid - solid)?
Trisha Rice at the ESEM applications lab might be able to provide this. She can be reached at either trice-at-feico.com or trice-at-electroscan.com (I'm not sure if this last address is still valid).
Dear Randy, I have just a few points to add to Steve's excellent advice.
1. "It is too dim to focus" - then i) increase the emission current, many people run at too low a current and make the task of optimising the instrument settings much more difficult ii) increase the kV which would increase the intensity too ii) re calibrate the photographic system to allow a focus intensity that is suitable for most operators
I often use LoDose film, which is more than an order of magnitude more sensitive than SO163, so I can barely see the beam on the screen. Fortunately, we have an intensified CCD, which is so sensitive that I can still focus. In fact, using the minimum-contrast method works very well with this system, since the contrast can be electronically enhanced.
6. To focus at low magnifications try removing the objective aperture and focus without a binocular, or a wobbler, looking for the very strong minimum contrast effect.
Furthermore, I discovered that inserting the diffraction aperture (with the objective aperture out) allows this method to be used at up to 63kx (and possibly higher). Since minimum contrast can easily be found within one second, this is well suited to radiolabile specimens.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
This is a different procedure from the Formvar-coated grids that you are using but it's one of my favorite journal articles/techniques - J. C. Rowley and D.T. Moran, A Simple Procedure for Mounting Wrinkle-Free Sections on Formvar-Coated Slot Grids. Ultramicrotomy 1 (1975) 151-155. They use Formvar-coated aluminum supporting racks. We use this method for slot grids all the time rather than Formvar-coated grids. good luck, Beth } TO be more specific, any advice on using formvar coated slot grids for } serial sections. They seem to crinkle, or get blobby, right on the vessel } I want to look at.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I do quite a bit of serial sectioning and would like to offer you the following advice:
1) Cut your sections as small as possible, they can be as much as 1mm wide, but try to make them something like 0.1mm high. This will allow you to get you as many as 20 sections on a 2mm slotted grid.
2) Get a bunch of locking tweezers and from the time you pick up the grid until you put it in the microscope, do not put the grid down onto filter paper (as you would for a normal grid). So, after you pick up the ribbon of sections, leave the grid in the tweezers to dry, and then when you post stain the grids, leave them in the tweezers to dry. If you put them down onto filter paper the formvar may rupture.
3) Maintain a positive attitude, and if things aren't going your way, stop and come back to it another day (the blocks will always be there for you, but if you get burned out you won't be there for the blocks).
Best of Luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
NATIONAL INSTITUTES OF HEALTH Bethesda, Maryland Division of Bioengineering & Physical Science Supramolecular Structure & Function Lab.
Biologist GS9/GS11/GS12 trained in life sciences and/or physical sciences at BS or MS level with solid technical experience in thin-section transmission electron microscopy. Experience in advanced techniques in analytical electron microscopy and structural biology (cryosectioning, high-pressure freezing, and macromolecular preparation methods) is desirable, although on-the-job training is possible for an experienced microscopist. PROOF OF U.S. CITIZENSHIP IS REQUIRED for this appointment.
For further information please contact:
Dr. Richard Leapman Division of Bioengineering & Physical Science Bldg. 13, Rm. 3N17 National Institutes of Health Bethesda, MD 20892 Tel: (301) 496-2599 FAX: (301) 496-6608 E-mail: leapman-at-helix.nih.gov
Applications (with curriculum vitae or federal employment form SF-171) should include the reference number ORS-01-0044, and should be post marked by April 9, 2001 and sent to:
Attention: Mr. Harold Atkins National Institutes of Health ORS Human Resources Office 31 Center Drive Msc 2157 Bldg 31, Room 4b41 Bethesda MD 20892-2157
Many thanks to everyone who responded to my question about focusing. Although my actual question was answered almost immediately, the ensuing replies were very enlightening. In EM work, it seems, even the "simplest" things have dimensions that aren't always obvious.
My initial question arose from the fact that I was curious about something I had always done as a matter of course because I had been taught to do it that way. That is, I have spent years focusing TEM's with the beam at the crossover point because I believed that I'd been taught to do so because of some property of electron "optics" that made it the preferred method to ensure best focus. Turns out, though, that the reason was something else entirely, involving an earlier generation of microscopes that simply had a beam that was too dim for focusing at the intensity needed for recording an image.
It's always educational to question old assumptions. Embarrassing, too, sometimes....
Thanks again, listers.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Technically, re-embedding paraffin sections for EM is perfectly feasible. The way I have done it is to separate the sections from slides after re-hydration and before further fixation with glutaraldehyde and osmium. Since both glutaraldehyde and osmium treatment cause sections to shrink and be brittle, sections might get further torn if they are still attached to the slides. Another step you should be careful about is to keep sections flat when applying glutaraldehyde and osmium. This ensures sections not to curl or wrinkle later. You can float your sections in 0.1 M phosphate buffer in a Petri dish, and let sections lie flat on the bottom. Then you gently remove buffer and add fixative with a glass pipet without disturbing the sections. Do not shake sections for a while after adding fixative. To flat-embed the sections, you can sandwich sections that have been fully infiltrated with resin between two sheets of Aclar film, then place this sandwich between two glass slides and apply some weight on top while polymerizing the resin. As someone already pointed out, even though you can re-embed paraffin sections for EM, ultrastructural will not be adequate anymore. This is particularly a problem for examining brain tissue in which membrane outline is essential for identifying different neuronal elements. However, if the tissue is precious, I would say it is still worth trying. We work with post-mortem brain tissue often. Sometimes the brains were over twenty-four hours post-mortem, fixed with only formalin and stored in cryoprotectant in a freezer for a long time. I am often surprised how much information we can still obtain from such tissue at EM level. I would be happy to help you to evaluate your micrographs if you like.
Good luck, Tina, let me know if you need more detailed answers.
Hong
============== Hong Yi Supervisor Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
---------- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: TEM - immunolabeling } Date: Wed, Mar 7, 2001, 2:55 PM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have two questions for you immunolabeling experts. } } First, a client has some precious and irreplaceable histological sections } of human hippocampus that have been labeled with ABC - DAB. Will it be } possible for me to de-paraffinize the sections, expose them to osmium } vapor and re-embed them in resin on the slide and pop them off and } resection them and see the DAB precipitate? Anyone have a protocol that } has worked? } } Second, these researchers plan to use Vibratome sections of mouse brain } and immunolabel them for light microscopy using an ABC kit. I have done } on-ultrathin-section colloidal gold immunolabeling for TEM. However, for } this new project we would like to try pre-embedding labeling of 50-60 } micrometer Vibratome sections, then embed and resection them for } TEM. Knowing what works for light microscopy, we'd like to use } streptavidin/colloidal gold or whatever for TEM visualization. My question } is, of course, does anyone have a favorite protocol they would be willing } to share? How do I keep the sections from curling? If we stick them on a } glass slide to embed in resin and (hopefully) pop off later for } resectioning, when and how do I get the to adhere? } } Mahalo! } } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
Regarding your question about pre-embedding immunolabeling of vibratomed brain sections, the type of labeling to use depends on the questions the researchers are trying to answer. Immunoperoxidase gives greater labeling depth, but limited spatial resolution. If they simply want to know what neuronal elements are labeled, then immunoperoxidase is fine. Otherwise, imunogold should be considered because it provides much more precise localization.
The methods for pre-embedding immunogold labeling using vibratomed brain sections have been very well documented. One of the first publications was by Chan, Aoki and Pickel (1) at Cornell University in 1990, shortly after ultrasmall gold conjugates were introduced. Many brain researchers have adapted their protocol. The characteristics of this protocol are the use of acrolein in the fixative for perfusion, using little or no detergent treatment for permeabilization and using a 1 nm colloidal gold conjugated IgG secondary probe in conjunction with a light insensitive, low viscosity, but fast reacting silver enhancement reagent. I have seen many beautiful results using this protocol. However, the particle size tends to be big, and irregular in shape. The cause of this is mainly in the silver enhancement reagent. In 1999, a new EM grade silver enhancement reagent was introduced to the market. Because of the improved enhancement homogeneity in particle size and shape, this silver enhancement reagent allowed us to conduct for the first time the pre-embedding double immunogold labeling in brain tissue using only ultrasmall gold conjugates (2).
To avoid a lengthy posting, I will send you the protocol, along with images off-line.
1. Chan J. Aoki C. Pickel VM. (1990) Optimization of differential immunogold-silver and peroxidase labeling with maintenance of ultrastructure in brain sections before plastic embedding. Journal of Neuroscience Methods. 33(2-3):113-27,
2. Hong Yi, Jan L.M. Leunissen, Ge-Ming Shi, Claire-Anne Gutekunst, and Steven M. Hersch (2001) A Novel Procedure for Pre-embedding Double ImmunogoldSilver Labeling at the Ultrastructural Level J. Histochem. Cytochem. 2001 49: 279-284.
Hong
============== Hong Yi Supervisor Emory University School of Medicine Neurology Microscopy Core Laboratory Rm 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
---------- } From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: TEM - immunolabeling } Date: Wed, Mar 7, 2001, 2:55 PM }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have two questions for you immunolabeling experts. } } First, a client has some precious and irreplaceable histological sections } of human hippocampus that have been labeled with ABC - DAB. Will it be } possible for me to de-paraffinize the sections, expose them to osmium } vapor and re-embed them in resin on the slide and pop them off and } resection them and see the DAB precipitate? Anyone have a protocol that } has worked? } } Second, these researchers plan to use Vibratome sections of mouse brain } and immunolabel them for light microscopy using an ABC kit. I have done } on-ultrathin-section colloidal gold immunolabeling for TEM. However, for } this new project we would like to try pre-embedding labeling of 50-60 } micrometer Vibratome sections, then embed and resection them for } TEM. Knowing what works for light microscopy, we'd like to use } streptavidin/colloidal gold or whatever for TEM visualization. My question } is, of course, does anyone have a favorite protocol they would be willing } to share? How do I keep the sections from curling? If we stick them on a } glass slide to embed in resin and (hopefully) pop off later for } resectioning, when and how do I get the to adhere? } } Mahalo! } } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
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A user has encountered an interesting problem . When cells are scraped and pelleted and embedded directly in epoxy they appear OK under the scope. When they are enrobed in agarose and then embedded the appearance becomes variable, although most cells appear very dense with contrast between organelles very low. This variability in appearance can be on the same grid square. Any ideas? Thanks Bruce Bruce Cutler Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id NAA02400 for dist-Microscopy; Mon, 12 Mar 2001 13:48:03 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id NAA02392 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 12 Mar 2001 13:47:33 -0600 (CST) Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu [134.193.71.1]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id NAA02384 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 12 Mar 2001 13:47:22 -0600 (CST) Received: by email.exchange.umkc.edu with Internet Mail Service (5.5.2653.19) id {FCF6B0PG} ; Mon, 12 Mar 2001 13:45:30 -0600 Message-ID: {95A711A70065D111B58C00609451555C01CA7611-at-UMKC-MAIL02} "'Microscopy-at-sparc5.microscopy.com '" {Microscopy-at-sparc5.microscopy.com}
May be it's not really a phase transition, but you can find pictures of dissolution and crystallization of table salt at our web page: http://www.umkc.edu/dentistry/microscopy/Esemimg.htm
Vladimir
-----Original Message----- } From: Edmund Gierlik To: Microscopy-at-sparc5.microscopy.com Sent: 3/9/01 10:24 AM
Email: gierlik-at-delta.sggw.waw.pl Name: Edmund Gierlik
Organization: Electron Microscopy Lab.
Education: professor
Location: Warsaw, Poland
Question: Can anybody show the source of nice presentation of ESEM application in a phase transition investigation (water - ice, liquid - solid)?
Dear listers, I have a customer who wants TEM on a section of cellulose that is coated with a layer of Fe2. I have no idea about how to go about accomplishing this. He wants to see where the Fe2 is located in the cellulose matrix, and how deep it has penetrated. The sample is a layer of cellulose about 1/2 mm thick with a visible layer of Fe on the surface and it's in submerged in water. Any ideas out there? Thanks so much,
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
After several requests, I've posted a measured schematic of my transmitted electron adapter, which can be reached via a link from the page mentioned previously:
http://www.mta.ca/~jehrman/ted.htm
I had the adapter made of aluminum and brass (just because the machinist likes to work with these). The angled surface below the specimen grid is polished brass, and as mentioned by several respondents, could be plated/coated/covered with something that generates more secondaries. Signal, however, has not been a problem in our applications - more than enough even at the smallest spot sizes.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I have a LKB Ultrotome V which has given many years of faithful service to me and my students until today, when the band that raises and lowers the cutting arm snapped. I am in the middle of the semester class on EM techniques and hence am a bit desperate to get it repaired. Does anyone know where I might find a replacement part for this - and is it possible to replace and adjust it myself? There are no instructions in my manual for this procedure; are there any out there somewhere? Is there any company that will service this instrument? I am located in New England.
Many thanks in advance for any leads.
Sincerely,
Dick Briggs Biology Department Smith College Northampton, MA 01063
This notice is to invite you to attend and participate in the 28th Annual Meeting of the Microscopical Society of Canada. This three day meeting will be held at the University of New Brunswick, Fredericton, New Brunswick, Canada on 6-8 June, 2001. The Scientific Program features invited speakers from the environmental sciences, plant sciences, material sciences, EELS applications, confocal microscopy and magnetic resonance imaging (MRI). Workshops will focus on specimen preparation techniques for both the biological and material sciences, including ultramicrotomy, polishing methods, high-pressure freezing techniques and a workshop on preparation of digital images for publication. Vendors and manufacturers will be demonstrating the latest developments in the field of microscopy. Presentations can be in either platform or poster format and the Abstract Deadline is March 30, 2001. The deadline for Pre-registration is April 30, 2001. All information and required forms for registration and submission of abstracts are available at the conference website at: http://www.unb.ca/msc2001. ************ Keynote Speakers and topics include: Dr. Bruce Balcom, Department of Physics, University of New Brunswick. Title of talk: "Magnetic Resonance Imaging of Materials". Dr. Ron Gronsky, Materials Science and Engineering, University of California, Berkeley. Title of talk: TBA. Dr. Michael Hochella, Dept. of Geological Sciences, Virginia Polytechnic Institute & State University, Blacksburg, Virginia. Title of talk: "Contributions of Microscopy to the Environmental Sciences: From Bacteria to Atoms" Dr. Richard Howard, DuPont Agricultural Products, Experimental Station, Wilmington, Delaware. Title of talk: "Trends in imaging fungal pathogens for understanding the cell biology of plant disease" Dr. Peter Ottensmeyer, Medical Biophysics, University of Toronto. Title of talk: "3-D Reconstruction of Macromolecules from single-particle STEM images: Transmembrane signalling of the insulin receptor"
Invited Speakers include: Mr. Jason Davis, Medical Biophysics, University of Toronto. Title of Talk: "EM of Chromophores: Very Low Energy-Loss Imaging of Green Fluorescent Protein and other Coloured Denizens." Dr. Gianluigi Botton, Materials Technology Laboratory, Natural Resources Canada. Title of Talk: "Analytical TEM of Macrostructures and Nanostructures". Dr. Elaine Humphrey, Biosciences EM Facility, University of British Columbia. Topic: "Confocal Microscopy" Dr. John McCaffrey, Institute for Microstructural Sciences, National Research Council Canada. Title of talk: "Structural Characterisation of InAs/GaAs and InAs/InP Quantum Dots by TEM" Dr. Doug Perovic, Dept. of Metallurgy & Materials Science, University of Toronto. Topic: "EM of Semiconductors". ************ Fredericton, New Brunswick is a three hour drive north from Bangor, Maine and an eight hour drive from Boston or Montreal. June is a beautiful time in New Brunswick and you can find interesting things to do and see through the conference website tourism links.
We turned water to ice on plant surfaces using the Peltier stage. We did not take any nice pictures (I have a few ugly ones!). It was an idea re ice nucleation on plants for a student project idea that we did not follow up.
Dave
On Mon, 12 Mar 2001 13:45:29 -0600 "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } May be it's not really a phase transition, but you can } find pictures of dissolution and crystallization of } table salt at our web page: } http://www.umkc.edu/dentistry/microscopy/Esemimg.htm } } Vladimir } } -----Original Message----- } } From: Edmund Gierlik } To: Microscopy-at-sparc5.microscopy.com } Sent: 3/9/01 10:24 AM } Subject: ESEM-Need presentation of phase transition. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Email: gierlik-at-delta.sggw.waw.pl } Name: Edmund Gierlik } } Organization: Electron Microscopy Lab. } } Education: professor } } Location: Warsaw, Poland } } Question: Can anybody show the source of nice presentation of ESEM } application in } a phase transition investigation (water - ice, liquid - solid)? }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need to fix live cells (mainly blood cells) on a microscope glass slide to prevent them from movement but to maintain their viability for 1-3 hrs. Also it should be non-stained with any dye cells in mono-layer. Material to fix cells should be optically transparent in 500-700 nm. Room temperature is asssumed.
Any input about applicable technologies (gels etc) with references about suppliers of materials that are involved and sources for description of those techniques will be highly appreciated.
Thanks in advance
Dmitri Lapotko, Ph.D.
Luikov Heat and Mass Transfer Institute 15, Brovka Street Minsk, 220072 Belarus
----- Original Message ----- } From: "Patton, David" {David.Patton-at-uwe.ac.uk} To: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} Cc: "'Edmund Gierlik '" {gierlik-at-delta.sggw.waw.pl} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 13, 2001 4:07 AM
} I am getting confused about light sources and Ca ratio imaging. I } have talked to people who use standard Hg arc sources and seem to } have no problems. Others seem to feel that you need a stabilized } power supply or a Xenon source for stability (see below). Are we } talking about line voltage variations (which a line } conditioner/voltage regulator ought to deal with) or actual arc } source variation? Any opinions out there? Thanks- Dave
} From my observations of using both types of light source, the Xenon seems } far more stable with time. You don't notice the fluctuations that you get } with Hg bulbs. This is important for quantification, such as dual excitation } ratio methods (eg. Fura-2). } } Stephen H. Cody, } Colon Molecular and Cell Biology Laboratory, } Ludwig Institute for Cancer Research, } Post Office Royal Melbourne Hospital, } Parkville, Victoria 3050, Australia. } } Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 } email: stephen.cody-at-ludwig.edu.au } www.ludwig.edu.au/confocal } } } } ---------- } } From: Tom Phillips[SMTP:PhillipsT-at-MISSOURI.EDU] } } Reply To: Confocal Microscopy List } } Sent: Wednesday, 28 February 2001 2:08 } } To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU } } Subject: Re: Xenon lamps } } } } All true but I believe one disadvantage is the peak illumination is } } less for some fluorochromes such as DAPI. We see a difference in our } } two systems that have Hg or Xe. } } } }
--
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Greetings, Two standard points. These are not about power output but do relate to the choice of arc type (archtype??) which seemed to be on the questioner's mind. Forgive me if this is too obvious.
One is that the output of a xenon arc is much more uniform across the spectrum compared to the steep peaks of the mercury arc. For ratios with different excitation wavelengths, in principle difference in intensity between the two wavelengths doesn't matter, but in practice when the intensities differ alot, there can be problems. Therefore, if you are going to be working with a variety of wavelength pairs (lots of different dyes and applications) the continuity of the xenon will probably be helpful; whereas, if you will stick to a single pair of wavelengths, or ratio emitted light, then even spectral quality might not matter.
The other issue is that xeon arcs tend to give off lots more ozone than mercury arcs and so usually require significant venting. This can be a nuisance depdending on the lay of the land, etc.
As ever, Tobias Baskin
} } } I am getting confused about light sources and Ca ratio imaging. I } } have talked to people who use standard Hg arc sources and seem to } } have no problems. Others seem to feel that you need a stabilized } } power supply or a Xenon source for stability (see below). Are we } } talking about line voltage variations (which a line } } conditioner/voltage regulator ought to deal with) or actual arc } } source variation? Any opinions out there? Thanks- Dave } } } } } From my observations of using both types of light source, the Xenon seems } } far more stable with time. You don't notice the fluctuations that you get } } with Hg bulbs. This is important for quantification, such as dual excitation } } ratio methods (eg. Fura-2). } } } } Stephen H. Cody, } } Colon Molecular and Cell Biology Laboratory, } } Ludwig Institute for Cancer Research, } } Post Office Royal Melbourne Hospital, } } Parkville, Victoria 3050, Australia. } } } } Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 } } email: stephen.cody-at-ludwig.edu.au } } www.ludwig.edu.au/confocal } } } } } ---------- } } } From: Tom Phillips[SMTP:PhillipsT-at-MISSOURI.EDU] } } } Reply To: Confocal Microscopy List } } } Sent: Wednesday, 28 February 2001 2:08 } } } To: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU } } } Subject: Re: Xenon lamps } } } } } } All true but I believe one disadvantage is the peak illumination is } } } less for some fluorochromes such as DAPI. We see a difference in our } } } two systems that have Hg or Xe. } } } } } } } } -- } } ************************************************************ } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269-3125 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html } ************************************************************
It could be a problem with penetration through the agar if cells are fixed in glutaraldehyde after the agar is added. Glut crosslinks the agar so that other solutions, including osmium and washes, can't penetrate properly. (I know this from experience.) We rinse the monolayer briefly and gently with warmed buffer (pour on, swirl, pour off). The buffer should be 300 mOsm (check with an osmometer) and pH 7.2-7.4 (check with a pH meter). Pour on the glut in buffer and let sit about 3 min (no more than 5); scrape, pellet cells in the glut and let sit for another 30 min. The pellets should be small (2 mm or so). If pellets are large, you can loosen them with the applicator stick so that the glut can get around to the underneath side. Just try not to disturb the clump too much. Remove glut with pipette. Scrape out pellet with flattened end of applicator stick. Enrobe with agar and then wash and stain as you would a piece of tissue. If there's a lot of excess agar, you can trim it away with a razor blade. This keeps them together and makes life much easier.
Some folks suspend cells in warm agar or gelatin before fixation and spin them down in the agar or gelatin and then fix. We have had variable staining this way, particularly if there is a lot of excess agar or the agar solidifies before the cells get to the bottom of the tube. If you can get a tight pellet by centrifuging fast in a warmed centrifuge (or jacketed tube) before the agar solidifies so that the layer of agar around each cell is thin, it will work. Do cut away the excess agar at the top of the tube, and make cell blocks small.
We centrifuge in a microfuge tube that looks like this:
| | | | | | | | | | \ / | | | | U
(Sorry about the graphics.) I think they come from PGC Scientific (800 424 3300). Sorry, I don't have the catelog number; our bag has been emptied, and our catelog has been "borrowed." They are about 2 " long and 3 mm in dia; the inside of the very tip is the diameter of a paper clip. This info was provided a while back, and maybe you can find it in the archives.
Centrifuge at right angles so that the pellet doesn't get stuck on the slanted neck of the tube. Then cut off the very bottom (below the cells) and then cut off the tube above the cells. Push out the cell "log" with a straightened paper clip. Chop into smaller logs.
Good luck. Address below if further questions.
On 12 Mar 2001, Bruce Cutler wrote:
} Date: 12 Mar 2001 12:58:48 -0600 } From: Bruce Cutler {bcutler-at-eureka.idl.ukans.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Agarose enrobed cells } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A user has encountered an interesting problem . When cells are scraped and } pelleted and embedded directly in epoxy they appear OK under the scope. When } they are enrobed in agarose and then embedded the appearance becomes variable, } although most cells appear very dense with contrast between organelles very low. } This variability in appearance can be on the same grid square. } Any ideas? } Thanks } Bruce } Bruce Cutler } Director, Microscopy & Electronic Imaging Laboratory } University of Kansas } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Dear Mary Gail, Do you mean a layer of metallic iron? I think you should treat the cellulose as wood or plant tissue and fix and embed, then cut a cross-section as you would a plant stem. No staining necessary. You need a TEM or STEM with an EDX on it and it will easily locate the iron to the ten nanometer scale. At 03:05 PM 3/12/01 -0500, you wrote: } } Dear listers, } I have a customer who wants TEM on a section of cellulose that is coated } with a layer of Fe2. I have no idea about how to go about accomplishing } this. He wants to see where the Fe2 is located in the cellulose matrix, } and how deep it has penetrated. The sample is a layer of cellulose about } 1/2 mm thick with a visible layer of Fe on the surface and it's in } submerged in water. } Any ideas out there? } Thanks so much, } } Mary Gail Engle
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hello, I was looking at some piping in our lab while sitting on the windowsill and noticed a sign painted over. Looking closer it was a sign for asbestos insulation that has been 80% painted over. I was told earlier that there was no asbestos pipe insulation in the lab. I collected some dust samples from near the fraying pipe insulation, on top of shelves, floor, and the windowsill and imaged them in our SEM.
Can anyone with some experience with asbestos take a look at the images at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the images looks like a candidate for asbestos? I think only fd6, fd7, and fd8 are asbestos, and the rest of the fibrous materials imaged are cellulose binding agents used in the insulation.
I will examine the material by TEM and electron diffraction, but I was wondering if someone could post the techniques they use for sampling from the dust, and any other hints and techniques I should be aware of.
Thanks for your help, Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
We have some data from a Wyko optical profilometer which we would like to work with offline & import into MATLAB for some further analysis. The lab tells me that the Wyko data can be output in "OPD" format. Does anyone know how we can covert this to a format we can work with.
Thanks
Tim
------------------------------------------------------------------------- Tim E. Harper CEO CMP Cientifica Tel. +34 91 640 7185 Fax. +34 91 640 7186
Gordon's comments concern me on several levels. The SEM, especially without EDX, is not the best tool to identify asbestos fibers. The TEM and diffraction can be absolutely definitive. However, I'm unaware of any EPA or NIOSH protocol for taking dust to the TEM grid, though one could be developed easily enough for Gordon's purposes. Gordon described friable pipe insulation with an asbestos warning. Seems wiser to bring his administration in on this situation. Does the UC system have some in-house personnel that can run a PLM for him? The lack of such in house personnel or an office dealing with asbestos abatement would be surprising for such a large university. Even a commercial lab PLM is faily inexpensive. While the amount of dust given off the pipe, barring some regular banging on the pipe, is probably minimal, the wiser course of action is bringing in someone who is well trained in dealing with asbestos pipe insulation. It doesn't have to be removed, just managed.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, I was looking at some piping in our lab while sitting on the windowsill and noticed a sign painted over. Looking closer it was a sign for asbestos insulation that has been 80% painted over. I was told earlier that there was no asbestos pipe insulation in the lab. I collected some dust samples from near the fraying pipe insulation, on top of shelves, floor, and the windowsill and imaged them in our SEM.
Can anyone with some experience with asbestos take a look at the images at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the images looks like a candidate for asbestos? I think only fd6, fd7, and fd8 are asbestos, and the rest of the fibrous materials imaged are cellulose binding agents used in the insulation.
I will examine the material by TEM and electron diffraction, but I was wondering if someone could post the techniques they use for sampling from the dust, and any other hints and techniques I should be aware of.
Thanks for your help, Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron MicroscopeLab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id IAA07825 for dist-Microscopy; Wed, 14 Mar 2001 08:32:38 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id IAA07822 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 14 Mar 2001 08:32:08 -0600 (CST) Received: from meta.vwh.net ([192.41.39.216]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id IAA07815 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 14 Mar 2001 08:31:56 -0600 (CST) Received: from metabolix.com (cint.metabolix.vne.verio.net [199.103.134.10]) by meta.vwh.net (8.8.5) id HAA16348; Wed, 14 Mar 2001 07:30:00 -0700 (MST) X-Authentication-Warning: meta.vwh.net: Host cint.metabolix.vne.verio.net [199.103.134.10] claimed to be metabolix.com Message-ID: {3AAF8061.D94223EE-at-metabolix.com}
I am trying to develop a procedure to thin section plant leaves, stain them with Nile blue, and visualize the stained leaves by light microscopy.
I assume that a microtome would be the best way to perform the thin sectioning but have I have no experience with microtomes. Does anyone have suggestions on suitable microtomes to purchase for such a procedure? If so, where is the best place to purchase them?
Thank you.
-Kristi Snell
-- Dr. Kristi D. Snell Metabolix, Inc. 303 Third St. Cambridge, MA 02142 Fax: 617-492-1996 Phone: 617-492-0505 x 229
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
A few years back when I took an EDS course put on by NORAN, they said keeping track of those K-alpha: L-alpha ratios should give you a running record of your detector sensitivity. I've been doing that ever since, and you can actually see those ratios change over time, as your detector window gets dirty. Another trick they mentioned (though I've never tried it myself) is to run a spectrum on some ordinary Teflon tape (yes, the kind you use for plumbing repairs). Teflon, apparently, contains no oxygen, so the presence of O in the resulting spectrum can only be from ice buildup. Clever, no? I also routinely do the Full Width Half Max test whenever I do a calibration. Good luck.
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 14, 2001 10:40 AM
I do not recall any problems with salt/water imaging.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----------------------------------------------------------------------. } } } We turned water to ice on plant surfaces using the Peltier } stage. We did not take any nice pictures (I have a few } ugly ones!). It was an idea re ice nucleation on plants } for a student project idea that we did not follow up. } } Dave } Like Dave, I've sometimes taken pictures of ice/water etc in our ESEM, but ice is such a good insulator, (I guess), the images never seem to come out well. So, I've never saved any. Also, salt/water interactions can be difficult too, since the salt really seems to want to charge a lot, to the detriment of image acquisition. Yes, you can play around with KeV and all that to minimize these effects, but it's still a very challenging "shoot" to get useable images.
Frank Thomas Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia
Brian, We work in a UKAS accredited laboratory and have a Pentafet light element detector with rotating turret device for windowless and thin window analysis. I perform a test with a Ni sample every month using the detector in its windowless mode. We monitor the ratio of the counts in the Ni L and Ni K peaks (NiL/NiL+NiK) (knowing what the value should be immediately after reconditioning). When the monthly value falls below a certain predetermined value, we simply perform the reconditioning routine. I suppose if you don't have a light element detector then the Cu standard you suggest is much better for a decent sized L line. I find that the above works very well for tracking the detector's sensitivity over a period of time. Best of luck Martin Roe
What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
I'd say some of those fibers are definitely "candidates". They are also large enough that you could analyze by PLM. If you want to do the TEM for fun or for the experience: dust samples are usually suspended in water and filtered onto a PC or MCE filter. I'm sure someone will post the appropriate reference, I can't seem to find it here.
Matt
On Tuesday, March 13, 2001 10:37 PM, Gordon Vrololjak [SMTP:gvrdolja-at-nature.Berkeley.EDU] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I was looking at some piping in our lab while sitting on the windowsill } and noticed a sign painted over. Looking closer it was a sign for } asbestos insulation that has been 80% painted over. I was told earlier } that there was no asbestos pipe insulation in the lab. I collected some } dust samples from near the fraying pipe insulation, on top of shelves, } floor, and the windowsill and imaged them in our SEM. } } Can anyone with some experience with asbestos take a look at the images } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } images looks like a candidate for asbestos? I think only fd6, fd7, and } fd8 are asbestos, and the rest of the fibrous materials imaged are } cellulose binding agents used in the insulation. } } I will examine the material by TEM and electron diffraction, but I was } wondering if someone could post the techniques they use for sampling from } the dust, and any other hints and techniques I should be aware of. } } Thanks for your help, } Gordon. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
This is a general site for Design News magazine; search that site for the term "lasik" to find an uncritical article featuring the inventor of the laser-vision machine now marketed by Visx. Even though it's a puff piece, it contains a great deal of info on the process.
2 recommended against the surgery, citing the risk of bad results for small gain if you have good correction now with glasses or contacts. Here are sites they cited with good info and extensive warnings of potential problems, and some additional sites I found in my own searches:
http://www.americaneye.com/bbs3/
This is a bulletin board where you can read many personal comments.
http://www.surgicaleyes.com/
This is a site founded by people who had unsuccessful laser vision correction resulting in complications. Its major point is that success defined by Snellen eye chart acuity may be inadequate.
http://wakeup.to/lasik
The site's name should tell you its slant on the topic.
http://www-psy.ucsd.edu/~mm/eyeknowwhy/index.htm
Very well-organized site, balanced viewpoint, but a bit out of date (last updated 1999). Well worth a careful reading anyway.
Detailed study results comparing PRK and LASIK outcomes. 3 years old (1998, therefore procedures done at least a year earlier). 220 eyes in the study. Interesting point (for me): 11.8% of PRK eyes and 3.2% of LASIK eyes reported a loss of 2 Snellen lines or more of best spectacle-corrected visual acuity (in English, you can't see as well as before even with glasses).
A more recent study (1999) with better outcomes. Equipment and experience makes a significant difference; be aware the technology changes fairly rapidly. 2-line loss rates from 2% to 0.6% in larger groups (1000+ and 700+ eyes).
A very good Feb. 2000 presentation of FDA pre-market approval data for several laser systems. 2-line loss is down to 0.3% with one system, but they show glare and night-vision problems reported by ~10% of patients. Remarkable Snellen acuity results: 87% 20/20 post-op without lenses.
5 respondents to my query had actually had laser surgery. All ultimately reported being satisfied with the results. 4 had no side effects at all. 3 reported good to excellent results immediately with no side effects. One (57 years old, farsighted) reported better post-op vision than previous lens-corrected vision. 1 respondent cited trouble for almost 1 year after surgery (couldn't see TEM images properly, trouble adjusting astigmatism), and said he was "initially devastated". However, he said he adjusted and/or the problems corrected themselves after a year. He can now handle all imaging tasks, and was happier overall than with glasses.
My personal decision has been to wait and see. I'm still not satisfied with the fairly high reported incidence of glare and night-vision problems, which I think are related to limited correction diameters compared to the dilated pupil of a larger-than-average eye (mine are). The changes I saw over just a few years of clinical trial results convinced me this is still evolving technology. The risks, while small, are still too high for me to accept in a purely elective procedure.
Thank you for your patience with this long-winded summary. Hope the info is useful to some of you.
Rick Mott (still reaching for his specs in the morning)
Dear Gordon, As someone in an old building with asbestos floor tiling, asbestos fume cupboard lining and old technicians who like to machine asbestos, I have looked at lots of samples of asbestos insulation. First, when asbestos is used, it is usally 30 % of the materials by volume, so it is very evident. Second it is characterized by very long, fairly straight fibres that divide down to smaller and smaller fibres, through and beyond the range of the SEM magnification. By that critierion, none of your pictures look like they contain asbestos. It helps to have looked at a sample so you can recognise it. Third, I check all suspicious fibres by EDX. The crysotile asbestos used in most insulation has a distictive Mg-Si spectrum, with a little Ca and Fe, that isn't found in most minerals. If it has that signature, then I would suspect asbestos and do the TEM. At 07:37 PM 3/13/01 -0800, you wrote: } Hello, } I was looking at some piping in our lab while sitting on the windowsill } and noticed a sign painted over. Looking closer it was a sign for } asbestos insulation that has been 80% painted over. I was told earlier } that there was no asbestos pipe insulation in the lab. I collected some } dust samples from near the fraying pipe insulation, on top of shelves, } floor, and the windowsill and imaged them in our SEM. } } Can anyone with some experience with asbestos take a look at the images } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } images looks like a candidate for asbestos? I think only fd6, fd7, and } fd8 are asbestos, and the rest of the fibrous materials imaged are } cellulose binding agents used in the insulation. } } I will examine the material by TEM and electron diffraction, but I was } wondering if someone could post the techniques they use for sampling from } the dust, and any other hints and techniques I should be aware of. } } Thanks for your help, } Gordon. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hello All, I have the opportunity to obtain a Seiko SMI8300 focused ion beam machine. I can't find out very much about it (I didn't even know Seiko made them - I suspect they are usually native to Japan). Does anyone have any useful information or experience? Could I use one to make TEM specimens?
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
One last note: of the 4 respondents who enthusiastically recommended the procedure, all were in agreement on the importance of finding a highly experienced surgeon. Complication rates are inversely related to experience.
I've never tried this either, but doubt it should work for the stated reason. A layer of ice on the detector (actually the oxygen in the ice) will act mostly as an x-ray absorber rather than a source of O x-rays. In fact, the ice will be a poor absorber for O-K x-rays because the oxygen x-ray energy is below its absorption edge.
Larry Thomas Pacific Northwest National Laboratory Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: Frank Thomas Sent: Wednesday, March 14, 2001 7:30 AM To: Brian Wajdyk; Microscopy-at-sparc5.microscopy.com Subject: Re: EDS detector chracterization: sensitivity/efficiency
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Brian -
A few years back when I took an EDS course put on by NORAN, they said keeping track of those K-alpha: L-alpha ratios should give you a running record of your detector sensitivity. I've been doing that ever since, and you can actually see those ratios change over time, as your detector window gets dirty. Another trick they mentioned (though I've never tried it myself) is to run a spectrum on some ordinary Teflon tape (yes, the kind you use for plumbing repairs). Teflon, apparently, contains no oxygen, so the presence of O in the resulting spectrum can only be from ice buildup. Clever, no? I also routinely do the Full Width Half Max test whenever I do a calibration. Good luck.
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 14, 2001 10:40 AM Subject: EDS detector chracterization: sensitivity/efficiency
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow Microscopists, } } Fellow Microscopists, } } I want to track the performance of our EDS detector over time. We } currently } check: peak to background ratios via peak-to-tail of Mn k-alpha, } symmetry } via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha } and } L-alpha to known values, and resolution via Full Width Half Max. What I } don't have is a good way to determine sensitivity of the detector to } determine when to remove ice/hydrocarbon build up. In the past I used a } 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha } because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. } Unfortunately I no longer have this standard or the means to acquire a } new } one thanks to the slow down in the e semiconductor industry. My best } guess } would to be to clean the detector then use the k-alpha to l-alpha ratio } of } Cu. However I know that we have the best microscopists in the world at } our } disposal. Any suggestions for sensitivity tracking, or other useful } detector characterizations I may have missed, would be greatly } appreciated. } } Sincerely, } } Brian Wajdyk } Sr. Electron Microscopist } On Semiconductor } brian.wajdyk-at-onsemi.com } 602-244-4883 } }
Perhaps a Macintosh, computer guru could figure this one out.
We are capturing JPEG images from a standard (Sony, analog 470 line, NTSC) video camera using a program called Reel Eyes. The images are captured on a Mac 8500 via the built-in S-Video port and saved as Quicktime JPG's (as mandated by the Reel Eyes program). When we put these images on our server they now appear with ".bin" appended to the file name so that the new image is described as a "Application/x-macbinary". Mac folks can open the files after processing through Stuffit but PC folks are unable to deal with the files. Anyway, this is an additional step that should not be present.
We can remove this problem by re-opening each of the images in Photoshop and saving them without the Thumbnail (or preview). So, I conclude that Quicktime is always saving with the Thumbnails in place. Does anyone know of a way that we can prevent the thumbnails from occurring in Quicktime?
Any other suggestions would be most welcome.
Many thanks,
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
The problem is that your files are encoded as Macbinary (.bin) before being stuffed. Windows machines can't decode .bin files unless they have the windows version of Stuffit expander. Encourage them to get it.
To correct this you need to change a preference setting.
In Magic Menu or Dropstuff preferences, go to the "Cross-Platform" or "MacBinary" icon and choose "Never" (instead of "Smart"). There is no way to change this preference in the StuffIt Deluxe application.
The URL for this information is {http://www.aladdinsys.com/support/techsupport/mac/dlx/dlx611.html}
Happy capturing. Gordon Nord "My real job is Mineralogy"
"John J. Bozzola" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Perhaps a Macintosh, computer guru could figure this one out. } } We are capturing JPEG images from a standard (Sony, analog 470 line, } NTSC) video camera using a program called Reel Eyes. The images are } captured on a Mac 8500 via the built-in S-Video port and saved as } Quicktime JPG's (as mandated by the Reel Eyes program). When we put } these images on our server they now appear with ".bin" appended to } the file name so that the new image is described as a } "Application/x-macbinary". Mac folks can open the files after } processing through Stuffit but PC folks are unable to deal with the } files. Anyway, this is an additional step that should not be present. } } We can remove this problem by re-opening each of the images in } Photoshop and saving them without the Thumbnail (or preview). So, I } conclude that Quicktime is always saving with the Thumbnails in } place. Does anyone know of a way that we can prevent the thumbnails } from occurring in Quicktime? } } Any other suggestions would be most welcome. } } Many thanks, } } John B. } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ##############################################################
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
Nord Consultants 20594 Cornstalk Terrace Ashburn VA 20147
Yes, I did use Hitach 4500S and 3500N to study some coated fibers in the recent past years. The coating layers were up to three.
General speaking, I think this kind of preparation needs patient and do it properly at every step as the processing goes.
The steps I did for preparing the coated fibers for SEM are 1. Cut the fibers carefully into about 1 cm length of pieces 2. Arrange them in a longitudinal direction (do not hurt them) 3. Hold one end of the fibers by using a pair of tweezers 4. Apply thin M-Bond (16) on to the fibers Since the M-bond is so thin, it will go along with the fibers and surround almost every fiber very well. 5. Leave the fibers in the air (room temperature) overnight. Do not let the wet part touch any solid surface such as metal or microscope slide. Otherwise when it cures, it is very difficult to separate them. Another way to cure it is putting the fiber with the tweezers on to a hot plate at 130 C for one hour. Do not let the wet part touch the hot plate. 6. When the fibers are cured, embed them into epoxy to make a big enough piece so that you can hold and polish it for SEM. At this step, you have to avoid trapping air in the epoxy. Especially when a fabric of fibers is going to be embedded into epoxy, you have to use a pump to get rid of air bobbles before curing. You may choose a kind of epoxy which can be cured in couple of hours in air (room temperature) or on a hot plate for about 1 to 2 hours at 130 C. The epoxy I used is a kind of mixture of two parts. 7. Then you may follow the routing procedures for polishing SEM samples, i.e., grinding first then polishing, from coarse to fine carefully.
Finally if you are going to study a fabric, then you can cut a size of 1 x 1 cm2, and hold it with a tweezers at a corner, and apply a layer of M-bond (16) on both sides, put it onto a hot plate for curing. Then embed this piece of fabric into epoxy. Again, you have to pump the air out before curing.
Good luck.
Zhenquan Liu
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Zhenquan Liu (Dr.)
------------------------------------ Zhenquan Liu (Dr.) Arizona State University Center for Solid State Science Room PSA213 Tempe, AZ 85287 Tel (480) 965 4544 (o) (480) 775 7428 (h) Fax (480) 965 9004 (o) Email zhenquan.liu-at-asu.edu ------------------------------------
Hello, We have a Gatan Duomill IBT Model 600. We have been having some problems with it lately. When I turn on the IBT, the vacuum in the main chamber works fine. When pumping down the specimen chambers, the left side does not pump at all. The right side pumps to the proper level, but when you lower the sample into the chamber, it takes a while for the vacuum to recover. There is no vacuum instability during sample rotation. There is no arcing in the guns. The argon flow is at the proper level. We have done the following: -cleaned, regreased and changed o-rings in the Whisperlock Assembly, sample viewing port, sample chamber, vent/vac buttons, and ion guns -changed the cathode tubes -dusted out the main sample chamber -check the vacuum pumps and topped off oil in the roughing pump -DP seems to be running fine
Still it doesn't seem to help. We are lost. Any suggestions about what might be going wrong here? Thanks. Prad
Pradyumna Prabhumirashi Department of Materials Science Northwestern University Phone: (847)-491-7798 Fax: (847)-491-7820 http://vpd.ms.northwestern.edu/prad.htm
We have used the k:alpha:l-alpha ratio for copper and nickel for many years as a measure of ice buildup on our detectors. Either ratio is very sensitive to ice buildup. With single element standards you don't have to worry if the mixture of two elements in your standard has changed. For sources we use copper or nickel TEM grids which are cheap and available from all the EM vendors.
Regards, Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Brian.Wajdyk-at-onsemi.com on 03/14/2001 10:24:35 AM To: Microscopy-at-sparc5.Microscopy.Com cc:
Fellow Microscopists,
Fellow Microscopists,
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
Just got a call from a researcher on campus looking for service on a Reichert Frigo-Cut type 2800, s/n 0110117 microtome. I am not familiar with this piece of equipment (we don't do light level microtomy) and I do not know this gentleman but he had already contacted one northern California service and was quoted $130/hr p to p. He balked. I explained that I had been quoted as much from services in New Hampshire. If anyone has a resource for repairing one of these microtomes in the Northern California region I will pass the info back to this person.
Thanks in advance.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Thanks for all of your replies, We had someone from our physical plant come in, and they revealed that the colour of the painted over sign indicates asbestos - free - insulation. Wierd scare for me since the painter covered the most important part of the label - that the insulation was asbestos free, not asbestos. They'll come back to tape back up all of the loose insulation tomorrow.
Since one of the specimens I looked at is very similar in morphology to asbestos, they'll be doing some wipe tests and counts by polarized light microscopy for asbestos particles to be sure. It likely was left behind from when the original asbestos insulation was removed about 10 years ago or so.
I appreciate all of the replies on the thread and supportive messages. Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 14 Mar 2001, Mary Mager wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Gordon, } As someone in an old building with asbestos floor tiling, asbestos fume } cupboard lining and old technicians who like to machine asbestos, I have } looked at lots of samples of asbestos insulation. First, when asbestos is } used, it is usally 30 % of the materials by volume, so it is very evident. } Second it is characterized by very long, fairly straight fibres that divide } down to smaller and smaller fibres, through and beyond the range of the SEM } magnification. By that critierion, none of your pictures look like they } contain asbestos. It helps to have looked at a sample so you can recognise } it. Third, I check all suspicious fibres by EDX. The crysotile asbestos used } in most insulation has a distictive Mg-Si spectrum, with a little Ca and Fe, } that isn't found in most minerals. If it has that signature, then I would } suspect asbestos and do the TEM. At 07:37 PM 3/13/01 -0800, you wrote: } } Hello, } } I was looking at some piping in our lab while sitting on the windowsill } } and noticed a sign painted over. Looking closer it was a sign for } } asbestos insulation that has been 80% painted over. I was told earlier } } that there was no asbestos pipe insulation in the lab. I collected some } } dust samples from near the fraying pipe insulation, on top of shelves, } } floor, and the windowsill and imaged them in our SEM. } } } } Can anyone with some experience with asbestos take a look at the images } } at: http://wilfred.berkeley.edu/~gordon/webdust and see if any of the } } images looks like a candidate for asbestos? I think only fd6, fd7, and } } fd8 are asbestos, and the rest of the fibrous materials imaged are } } cellulose binding agents used in the insulation. } } } } I will examine the material by TEM and electron diffraction, but I was } } wondering if someone could post the techniques they use for sampling from } } the dust, and any other hints and techniques I should be aware of. } } } } Thanks for your help, } } Gordon. } Regards, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchg.ubc.ca } }
A few years ago, a EDS reparer explain me a test, which seems to be interresting, to check the "health" of a detector with UTW, but I never have tried it.
With a fluorine (CaF2) sample at 10 keV, You should have a ratio of one betwween F K alpha and Ca K alpha. When not, or when it diminish in the time (less F), you have ice (absorbtion effect of F by O2). (Why 10 kev? Is 10 keV not a bit high for F. It think it's a compromise between the window caracteristics and the primary energy).
Same thing with quartz (SiO2), at 5 keV, and than, if O diminish, you have oil on the window (absorbtion effect of O2 by C).
As I said, I never tried this test, the CaF2 and SiO2 samples are in my drawer, waiting for polishing. I suppose that geologist should have an advice about that, but of course they don't often work at 5 keV for EDS.
I usually use a aluminum sample holder, with a piece of carbon adhesive tape and a copper foil, and I take a spectrum with a quarter of the SEM screen on the copper, a quarter on the carbon, and the half on the aluminium,at low mag, and at 5 keV. I adust the probe current to a preset value (typically a few nA, and each time the same value) mesured on the carbon tape (less SE and BSE, better would be a Faraday cup). So I see if the sensitivity on C K, Cu L and Al K seems to diminish from on mesure to an other. In such condition you'll see that the count rate (and of course, the spectrum shape) is very sensitive to window or detector contamination (with allways the same probe current, time constant, etc.). You can imagine similar test with for exemple, Vandium L and silicon K, etc.
best regard
Jacques
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I was asked to perform double labelling of surface antigens on leukaemia cell cultures and cord blood progenitors.
In the past I used a pre-embedding protocol for single labelling which worked quite well.
I would like to continue with a pre-embedded protocol, but both primary antibodies are raised in the same animal and I wonder how I can block unspecific labelling in that case.
(I have a protocol for post-embedded double staining with primary AB raised in the same animal, but would prefer to use that as a last resort)
So far I used a 10 nm gold F(ab)2- AB - if I use a 5nm gold F(ab)2 AB will I be able to distinguish them in the TEM?
I would appreciate your input very much.
Many thanks
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
I have a technique that I use with clients on a wide range of fibres which takes very little time.
1. Drill two or three holes through two SEM stubs placed face to face. 2. Pass the fibres through the holes. 3. Fix in place with a water based carbon solution, make sure the fibre/stub interface is well wetted. 4. When fully dry place in liquid nitrogen CARE!!. 5. When the bubbling stops lift out CARE!! 6. Place on an insulating surface and with a knife strike the interface between the two stubs - the system will fracture. 7. When dried off you have two sets of surfaces to look at in LM or SEM.
It works great on a number of differing media other than fibres, the only snag is they must not be affected by the water base.
Steve Chapman Senior Consultant Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I imagine that the problem arises when you transfer the files to the server. I'll assume that you are using some FTP program such as Fetch. In the upload preferences, make sure that you are transferring non-text data in the "raw data" format, as opposed to MacBinary, etc. Also, make sure that the FTP doesn't append a ".bin" to raw data.
Remember you still need to have a ".jpg" at the end of the filename or else the Windows boxes still won't know how to handle the file (sigh...). There are numerous utilties on the Mac to batch process the renaming of files for cross-platform compatibility.
Hope this helps, John B.
} } Perhaps a Macintosh, computer guru could figure this one out. } } We are capturing JPEG images from a standard (Sony, analog 470 line, } NTSC) video camera using a program called Reel Eyes. The images are } captured on a Mac 8500 via the built-in S-Video port and saved as } Quicktime JPG's (as mandated by the Reel Eyes program). When we put } these images on our server they now appear with ".bin" appended to } the file name so that the new image is described as a } "Application/x-macbinary". Mac folks can open the files after } processing through Stuffit but PC folks are unable to deal with the } files. Anyway, this is an additional step that should not be present. } } We can remove this problem by re-opening each of the images in } Photoshop and saving them without the Thumbnail (or preview). So, I } conclude that Quicktime is always saving with the Thumbnails in } place. Does anyone know of a way that we can prevent the thumbnails } from occurring in Quicktime? } } Any other suggestions would be most welcome. } } Many thanks, } } John B. } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ##############################################################
At 01:26 PM 3/14/01 -0600, John J. Bozzola wrote: } The images are captured on a Mac 8500 via the built-in S-Video port and saved as Quicktime JPG's (as mandated by the Reel Eyes program). When we put these images on our server they now appear with ".bin" appended to the file name so that the new image is described as a "Application/x-macbinary". Mac folks can open the files after processing through Stuffit but PC folks are unable to deal with the files. Anyway, this is an additional step that should not be present. } We can remove this problem by re-opening each of the images in Photoshop and saving them without the Thumbnail (or preview). So, I conclude that Quicktime is always saving with the Thumbnails in place. Does anyone know of a way that we can prevent the thumbnails from occurring in Quicktime?
The Mac's file system is slightly different than that on a PC or a Unix server. Mac files have two parts, known as forks. The icon or thumbnail image resides in the "resource fork". The regular JPEG image data that a PC expects is in the "data fork." The resource fork also holds the Mac's association between the file and the program that made it, while the PC uses the file extension for that purpose.
MacBinary files are a way of wrapping up the data and resource forks into a single file, such as for transmission via an e-mail attachment, ftp site, etc. or in your case, handy for putting on a server that doesn't speak with forked tongue.
You didn't say much about your server. If it's a WinNT server, your admin can install some AppleTalk extensions that can make PC {-} Mac file exchange a bit easier. They automatically manage the resource fork, hiding it for PC programs but allowing networked Macs to store and access it.
You also didn't say much about which PC programs you were using. I don't know of any that handle MacBinary files. (I've written a few that did, once upon a time, but they're obscure.) You may be forced to either strip the files on the Mac, or use a utility on the PC side to convert the MacBinary files to data-only files.
I don't think Quicktime thumbnails are the problem, I think it's a MacBinary problem.
Question: I have been analyzing stress around STI (shallow trench isolation) in DRAM. In general, stress is higher at bottom corner and top corner of STI, which is probed by my data. But, the strange thing is that HOLZ line split and blurring occurs in CBED pattern at the bottom of FOX (field oxide). I know that the split and blurring is the sign of high stress. And I don't think that stress is higher at the bottom of FOX. So, my question is what makes line split and blurring except stress. Have you seen that kind of phenomenon? Thank you for your answer.
If you go to our new website at www.southbaytech.com, you will find a listing of technical papers that you can request. There are several there dealing with coated fibers. Please take a look. If any look interesting, submit a request and we will send them out to you at no charge.
When you get to the site, click on Applications support then technical papers. There are a lot of papers there, so the easiest way to find the ones you are interested in is to use the "find on page" function typically found under your edit command. If you type in "fiber", you'll find some relevant papers.
I hope this helps.
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
The ion getter pump on the CM10 (12 and 20) does have a finite life and if your machine is 10 years old with the original IGP then I suspect that the pump could be finished. I assume that it will not pump down properly on the IGP but has a good penning gauge (P3) pressure.
The pump is not a standard Edwards pump as it has an extra port so that it can pump both the column and the gun. As I understand it the pump body is cut open to replace the plates and then rewelded before being leak tested and baked.
There are companies that will do this for you but you are without the pump while they do it, this may take two weeks or more. FEI should hold replacement pumps that they would swop in a day, more expensive but faster and more convenient. It depends on your budget and facilities.
Good luck, Ron
On Thu, 15 Mar 2001 14:08:46 +0100 Marco Arienti {marienti-at-tiscalinet.it} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have no big experience with Philips microscopes, but I need some } information. } } A CM10 installed about 10 years ago have some vacuum problems. } } Looks like the Ion Getter Pump is not working anymore. } } As far as I understood the problem may be that there is no more Titanium in } the pump. } } Is possible In this one to change the only grids (like in the Leibold pumps) } or the all pump have to be exchanged? } } It is a pump manufactured from Edwards, any info about the model? } } Thanks a lot. } } } } Marco Arienti } } www.electronrescue.com } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Could you recommend a water based carbon solution?
Dave
On Thu, 15 Mar 2001 04:56:05 -0500 Steve Chapman {PROTRAIN-at-CompuServe.COM} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi } } I have a technique that I use with clients on a wide range of fibres which } takes very little time. } } 1. Drill two or three holes through two SEM stubs placed face to face. } 2. Pass the fibres through the holes. } 3. Fix in place with a water based carbon solution, make sure the } fibre/stub interface is well wetted. } 4. When fully dry place in liquid nitrogen CARE!!. } 5. When the bubbling stops lift out CARE!! } 6. Place on an insulating surface and with a knife strike the } interface between the two stubs - the system will fracture. } 7. When dried off you have two sets of surfaces to look at in LM or } SEM. } } It works great on a number of differing media other than fibres, the only } snag is they must not be affected by the water base. } } Steve Chapman } Senior Consultant Protrain } For professional training in SEM, TEM and EDX world wide } www.emcourses.com }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need some info on liquid nitrogen dispensing devices. Our EM lab has four 20 liter dewars for liquid nitrogen storage. A piece of styrofoam (2 x 6) is attached to the lid. We would like to purchase a dispensing device because it would be easier to use and less waste of liquid nitrogen. Thanks in advance Karen Schlueter PWA, ARS, USDA EM Lab 1636 East Alisal St. Salinas, CA 93905 (831) 755-2847 FAX: (831) 755-2814 kschlueter-at-salinas.ars.usda.gov
As a materials scientist, I haven't had any experience doing TEM sample prep of biological samples, so I need some assistance in this.
I've been asked to look at in the TEM some dehydrated beef looking for the changes in sarcomere length etc due to the dehydration process.
I have at my disposal staining solutions of 0.5% RuO4, and 2% methanolic UA and need some procedures on how to go about preparing these specimens. (Things like cut the dehydrated sample into cubes, stain with this for how long at this temp then embed and microtome or such.)
Thanks in advance.
dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
I am looking to purchase a sputter coater ~$5000. I currently have a Polaron E5200 that is inoperable. I do Failure Analysis on semiconductors, probably a 6 inch chamber & Au/Pd target. Can anyone in the list give me some good tools that work for them.
Anyone familiar with BOC Scancoat Six?
Thanks in advance
Jim Arnold Senior Quality Technician / Failure Analysis Honeywell - Microelectronics and Technology Center Columbia, MD 21045 email: jim.arnold6-at-honeywell.com
For organic contamination, monitoring the ratio of TiKa to TiLa is good because TiLa is absorbed heavily by organic layers on the detector window. For ice, a spectrum from say calcium fluoride is good because again fluorine Ka is heavily absorbed by oxygen containing layers such as ice.
Carl Miller ---------------------- Forwarded by Norman C Miller/RES/Raytheon/US on 03/14/2001 02:00 PM ---------------------------
"Brian Wajdyk" {Brian.Wajdyk-at-onsemi.com} on 03/14/2001 09:40:51 AM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: Norman C Miller/RES/Raytheon/US)
Fellow Microscopists,
Fellow Microscopists,
I want to track the performance of our EDS detector over time. We currently check: peak to background ratios via peak-to-tail of Mn k-alpha, symmetry via Full Width Tenth Max, peak offset via centeroid of the Mn k-alpha and L-alpha to known values, and resolution via Full Width Half Max. What I don't have is a good way to determine sensitivity of the detector to determine when to remove ice/hydrocarbon build up. In the past I used a 80%Mn 20%Cu standard and looked at the ratio of Cu k-alpha to Mn l-alpha because at 10kV accelerating voltage, the peaks are nearly a 1:1 ratio. Unfortunately I no longer have this standard or the means to acquire a new one thanks to the slow down in the e semiconductor industry. My best guess would to be to clean the detector then use the k-alpha to l-alpha ratio of Cu. However I know that we have the best microscopists in the world at our disposal. Any suggestions for sensitivity tracking, or other useful detector characterizations I may have missed, would be greatly appreciated.
Sincerely,
Brian Wajdyk Sr. Electron Microscopist On Semiconductor brian.wajdyk-at-onsemi.com 602-244-4883
I am looking to replace my target on my sputter coater and would like to find out if there is an optimal thickness for the target. Is it material dependant? I had requested 2.5mil from the thickness of my old target but the vendor I spoke with will only source a minimum 4mil thick target. I am looking at Pt and Au/Pd. Will I have to increase current to get the same deposition rate for a thicker target?
TIA
David B Rose W. L. Gore and Associates 297 Blue Ball Road Elkton, MD 21921 410-506-2958
Many thanks to all of you who offered advice, assistance, consolation and even a loan of the part or a used instrument for parts. I was able to find a replacement ($100 - overpriced perhaps but worth every penny!) and will get it installed next week with the directions from another microscopy list reader.
Again my thanks to all and if anyone wants the names and addresses of some LKB parts dealers and/or service folks, I will gladly provide you with my new list off-line.
Some years ago I had Melins (510-234-5749) repair our Reichert 2800. They insisted on taking it to their shop and it needed a new compressor from Europe. The bill was over $2000. $130/ hour is a typical charge for this type of work in this area. When I the Reichert unit was destroyed in a water flood I purchased a Microm cryostat. It uses standard easily available refrigeration components. The cost and repair time is gresatly reduced. Good luck.
At 03:35 PM 3/14/01 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
No, thicker target = longer life, but it makes no difference to deposition rate, current etc. Chris
----- Original Message ----- } From: {"drose-at-wlgore.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 7:23 PM
No, thicker target = longer life, but it makes no difference to deposition rate, current etc. Chris
----- Original Message ----- } From: {"drose-at-wlgore.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 7:23 PM
G'day
I have an aging Jeol 840 and the monitor image is getting a bit dim and noisy to the extent that I have to use large spot sizes that are damaging some samples! Jeol tell me they cannot supply new tubes. Does anyone know of compatible tubes that could be utilised? Is there anything special about SEM monitor tubes?
Thanks Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
I have some difficulties to stick my protein (Mojor Sperm Protein, a basic protein from Nematode Sperm ) to the microscope slide, therefore, it is hard to perform further perfusion experiment. When I try to do so, I lose everything. Does anyone have experience in such a manipulation? Any suggestion is highly appreciated. Long
The CRTs can be rebuilt for about $600.00 to $800.00 USD by Richardson Electronics in Georgia.
I have their number & can give it to you offline.
Earl weltmer
----- Original Message ----- } From: "Dave Phelan" {emudp-at-mail.newcastle.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 15, 2001 4:35 PM
Try Richardson Electronics http://www.rell.com/ . Have CRT information handy (labels on the CRT) when calling Richardson Electronic. Good luck.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
G'day
I have an aging Jeol 840 and the monitor image is getting a bit dim and noisy to the extent that I have to use large spot sizes that are damaging some samples! Jeol tell me they cannot supply new tubes. Does anyone know of compatible tubes that could be utilised? Is there anything special about SEM monitor tubes?
Thanks Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
Hi: We are lookng to purchase a Zeiss 10C TEM, preferable in working condition but would consider one needing some work. Please contact us direct via e-mail or call 909 301-9130 (Los Angeles area) EMSI Peter Jordan
Dear All, I'm looking for circuit diags for J-TEC X-ray counting electronic units XM-XCU. These were supplied with some JEOL 733 'probes although most seem to have gone out with ORTEC. Can anyone help? Thanks, MAlc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
Both display and photo-monitor tubes can be recoated. We had our photo CRT recoated by Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if they're still in business, but they did a good job. JSIII
} Try Richardson Electronics http://www.rell.com/ . Have CRT information handy } (labels on the CRT) when calling Richardson Electronic. Good luck. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } } This message is made of 100% recycled electrons. } -----Original Message----- } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Thursday, March 15, 2001 10:28 PM } Subject: Jeol 840 viewing screen replacement } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Julian P.S. Smith III Dept. of Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x6427 (vox) 803-323-3448 (fax)
Does anyone use this resin and use it for antibody staining? We will be using this resin for embedding. Also any suggestions on etching. Thanks. Caroline Miller
I have a Tracor Northern TN-5402 EDS system mounted on a JEOL SEM. The monitor just went up in smoke and I need to a) find a replacement or b) try to replace the hardware. Since I don't have much money and we would like to replace the whole system in a couple of years I can't spend much to repair the monitor.
Does anyone have a monitor for sale/offer? Does anyone know of a replacement monitor available on the market? A standard BNC type of monitor does not work because of a proprietary modulation. Has anyone successfully upgraded the hardware minus the detector?
Thanks, Louie
-- Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
VISIT OUR WEB SITE: http://www.mbl.edu/ http://www.courses.mbl.edu/
Our Sorvall MT2B is having some problems, to say the least. I would very much appreciate any info. concerning service of this machine. We are located in Milwaukee, WI. I have contacted Boeckeler (new owners of RMC) regarding service. They informed me they are no longer providing service on these models. What other options do I have?
TIA Craig M. Klotz, BS,CT(ASCP) EM Tech II Neuromuscular Lab Medical College of Wisconsin cklotz-at-mcw.edu
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
Claudia, I would direct you to the March issue of journal of histochemistry and cytochemistry, they have published a procedure to double-label with gold at the pre-embedding stages, good luck Marge
Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation email: springett.margaret-at-mayo.edu
} ---------- } From: Claudia Hayward-Costa } Sent: Thursday, March 15, 2001 2:49 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: IEM: Double labelling of cells } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Microscopists, } } I was asked to perform double labelling of surface antigens on } leukaemia cell cultures and cord blood progenitors. } } In the past I used a pre-embedding protocol for single labelling } which worked quite well. } } I would like to continue with a pre-embedded protocol, but both } primary antibodies are raised in the same animal and I wonder how } I can block unspecific labelling in that case. } } (I have a protocol for post-embedded double staining with primary } AB raised in the same animal, but would prefer to use that as a } last resort) } } So far I used a 10 nm gold F(ab)2- AB - if I use a 5nm gold F(ab)2 } AB will I be able to distinguish them in the TEM? } } I would appreciate your input very much. } } Many thanks } } Claudia } } } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } 44(0)208 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
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} -----Original Message----- } From: Julian Smith III [mailto:smithj-at-Winthrop.edu] } Sent: Friday, March 16, 2001 12:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 840 viewing screen replacement } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Both display and photo-monitor tubes can be recoated. We had } our photo CRT } recoated by } Video display corp (Tucker, GA 800-241-5005) in 1993. I don't know if } they're still in business, but they did a good job. } JSIII } } } Try Richardson Electronics http://www.rell.com/ . Have CRT } information handy } } (labels on the CRT) when calling Richardson Electronic. Good luck. } } } } Vitaly Feingold } } Scientific Instruments and Applications } } 2773 Heath Lane, Duluth GA 30096 } } (770)232-7785 ph. } } (770)232-1791 fax } } } } This message is made of 100% recycled electrons. } } -----Original Message----- } } } From: Dave Phelan {emudp-at-mail.newcastle.edu.au} } } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } } Date: Thursday, March 15, 2001 10:28 PM } } Subject: Jeol 840 viewing screen replacement } } } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } } G'day } } } } I have an aging Jeol 840 and the monitor image is getting a bit dim } } and noisy to the extent that I have to use large spot sizes that are } } damaging some samples! Jeol tell me they cannot supply new } } tubes. Does anyone know of compatible tubes that could be } } utilised? Is there anything special about SEM monitor tubes? } } } } Thanks } } Dave } } } } } } } } } } } } Dave Phelan } } EM/X-Ray Unit } } University of Newcastle } } NSW 2308 } } AUSTRALIA } } Ph 02 4921 5667 } } Fax 02 4921 7019 } } emudp-at-mail.newcastle.edu.au } } } Julian P.S. Smith III } Dept. of Biology } Winthrop University } Rock Hill, SC 29733 } 803-323-2111 x6427 (vox) } 803-323-3448 (fax) } } }
As many of you know, I maintain the WWW pages for Project Micro which is dedicated to Educational Outreach to pre-college students. One of their projects is sand from around the world which is used in the classroom activities section.
Today I uploaded their latest web page documenting their sand collection administered by Joe Neilly of Abbott Labs . You can see this page at
I could not help to notice that their supply of sands from outside the USA has become sorely lacking. In particuliar
Europe, Africa and South America
are now all completely depleted. And
Australia and Asia
only have supplies from only one location left.
Can I suggest to members of this list particularly if you are not from the USA, that if you have the opportunity this would be an excellent and simple way that you can contribute to the education of our next generation of microscopist's. The sand is available to any educator, regardles of their affiliation and location.
All the information you need to contribute "sand" from your local beach is listed on that WWW page as well as the end of this message.
It will cost you a few $$ out of your pocket to send the sand , but remember, it is for a good cause. So spend a few minutes next time you drive past a beach and fill up a small plastic bag, and send it to Joe.
Cheers.... Nestor Your Friendly Neighborhood SysOp.
--------------------------- Here is an exerpt from the Sands WWW page
To Request Sand
Select the sands you would like from the list below and e-mail your request to joe.neilly-at-abbott.com. Include your selection (limit 6 per request), where the sands should be sent, and how you plan to use them. This collection was created for Microscopic Explorations but how you ultimately use them is up to you. Great stories and photos about how you used the sands are always welcome!
To Donate Sand
Sand donations are what keeps this collection going. All donations are appreciated and will be shared with many educators. To send your sand, fill a Ziplock sandwich bag half full and place it in another Ziplock bag. Mail your donation to:
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Dear Listservers, recently there was a discussion about the stability of LR-White in the EM-beam. One distribtion mentioned a polymerization of LR-White by microwave treatment. I don´t remember who sent this mail. I would be interested to test this procedure. Who knows how to do this kind of LRW polymerization? What is the set-up? How long does it take, how much Energy (Watt) is needed? Maybe the colleague who posted this mail remembers the discussion and is able to give some information.
Thanks in advance, with best regards, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________________ Alles unter einem Dach: Informationen, Fun, E-Mails. Bei WEB.DE: http://web.de Die große Welt der Kommunikation: E-Mail, Fax, SMS, WAP: http://freemail.web.de
Dear Michael: The following procedure for polymerization of LR-White has worked of me in my microwave oven: I will start with the infiltration phase: The wattage setting I used was about 700-750 watts.
1:1 95% ETOH / LR-White 50'C temperature restriction setting (trs) x2 for 10 minutes each.
100% LR-White 50'C (trs) x3 for 10 minutes each.
Polymerization in the oven: In this phase be sure to fill the beem capsule to the very top and cap off with parafilm to make a tight seal to prevent water from getting into the resin. Submerge the capsules in a water bath and place the temperature probe in the bath to monitor the temp.
The blocks will be firm and will cut very well. For more information on this technique you can contact Rick Giberson at TED PELLA CORP. USA using their toll free number, I don't know what that is in Germany! He can give you many more details about LR-White and the microwave oven. I understand that lesser wattage setting can be used to infiltrate and polymerize LR-White but I have not experimented with them as yet!
Good Luck
Ron Austin (Research Associate) Dept. of Pathology L.S.U. Medical Ct. Shreveport, LA 318-675-4775 rla-at-mindspring.com
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by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id LAA21224 for dist-Microscopy; Sun, 18 Mar 2001 11:59:42 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id LAA21217 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 18 Mar 2001 11:59:11 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id LAA21210 for {microscopy-at-msa.microscopy.com} ; Sun, 18 Mar 2001 11:59:00 -0600 (CST) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130303b6daa5368faf-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Colleagues
A number of people have reminded me about the growing problems with hoof and mouth disease around the world and that sending possible organic material from country to country should not be considered at this time.
I humbly acknowledge that you are all absolutely correct!
It simply did not occur to me to connect sand with this problem. I guess my physics background is just showing it's tunnel vision, since I think of sand as inorganic compounds and neglect to think of organic "contaminants" which might be included.
For the moment it would therfore certainly be prudent to hold off collecting and/or send and sand samples for the Micro Project, even if it can be documented that the samples have been properly sterilized. Let's wait until this potential problem is controlled. It is better to error on the safe side.
Thanks again, to everyone that pointed out my error.
Dear Microscopists, I am working with Ilex paraguariensis seeds. After dissection, with mechanical extraction of the endocarp, the micropylar endosperm is enveloped by a lignified cap. This one do not allow the cellular visualization, in clarified material (Herr´s method), of the endosperm because the yellow color of the lignin. Is there a non aggressive protocol for lignin extraction? Thank´s.
Rinaldo Pires dos Santos
Dr. Rinaldo Pires dos Santos - e-mail: rinaldop-at-uol.com.br Lab. of Plant Anatomy - Dept. of Botany - UFRGS Porto Alegre - RS - Brazil
Well, I stepped in it this time. I agreed to do some "gee-whiz" shots for my dentist of a tooth with a rather large amalgam filling. Only later did it sink in that the mercury might be a problem under vacuum and the beam. Has anybody out there done this? I'd be looking at the thing with gold coating an a fairly light beam (say 10 kV), but unfortunately, not with a cold stage. I've checked the archives and found one brave soul who said that he's done this on old amalgams. I can't really say how old this one is, but I'm pretty sure it wasn't made yesterday. Call me paranoid, but I'd like some more opinions before I do something really stupid!
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I am trying to evaluate some tools that perform automated TEM sample prep prior to FIB thinning. I am looking for some one that has experience using Sela microcleaving tools in conjuction with the TEMstation. I work with failure and material analysis in the IC industry so submicron accuracy and relaibility are important concerns of mine. Also, we will soon be facing chips with low K dielectrics which from what I have heard are extreme soft and hard to polish. I am not sure if the sawing technique they employ will be able to section this type of material.
If anyone on this list has any experience with these tools and can give me insight into their worth as a prep technique please get in touch with me.
We are interested in buying a liquid helium TEM cold stage. Has anyone had experience with it and how was it? Is there any other companies selling this apart from Gatan?
I would appreciate very much for any input on this.
Best regards Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
Does anyone have a step by step protocol for decalcifying bone for transmission electron microscopy using EDTA?
Thanks,
Kenn
Dr. Corazon D. Bucana, Ph.D. Mr. Kenneth Dunner, Jr. Department of Cancer Biology U.T. M.D. Anderson Cancer Center High Resolution Electron Microscopy Facility 7777 Knight Road, Box 173 Room SRB 1.660 Houston, Texas 77054 PH: 713-792-8106 FAX:713-792-8747
I am in need of a gas flow proportional counter (GFPC) for a JEOL 733 microprobe. Alternately, I may also need some W wire of the appropriate size to restring an existing unit. Thanks for any help/advice.
Ed Holdsworth General Mgr. SEMTEC Laboratories, Inc.
I have had good luck decalcifying bone for TEM using this EDTA protocol:
Fix 1-2 mm sized bone pieces for two days in 2.5 percent glutaraldehyde in 0.1 M Sorensen's buffer.
Decalcify on a shaker for 3-7 days in 7.5 % disodium EDTA, 2.5 % glut. in 0.1 M Sorensen's buffer. (pH the decal solution to physiological pH with NaOH.)
Change to fresh decal solution every couple of days.
Check for complete decalcification by taking before and after X-rays of the tissue.
Rinse twice in buffer
Post fix in 1 % osmium in buffer.
Rinse once with buffer, then once with ddH2O
en bloc stain for 1 hour with aqueous 3% uranyl acetate.
Dehydrate in a graded series of EtOH, infiltrate, and embed in epon.
Good luck, and I hope this helps.
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Mon, 19 Mar 2001 semcore-at-audumla.mdacc.tmc.edu-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have a step by step protocol for decalcifying bone for } transmission electron microscopy using EDTA? } } Thanks, } } Kenn } } } } } } } Dr. Corazon D. Bucana, Ph.D. } Mr. Kenneth Dunner, Jr. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } High Resolution Electron Microscopy Facility } 7777 Knight Road, Box 173 } Room SRB 1.660 } Houston, Texas 77054 } PH: 713-792-8106 } FAX:713-792-8747 }
I'm looking for a part no longer available from the manufacturer. We have a TMC Micro-g air table, and one of the black plastic parts that holds together the leveling valve (at the ends of the table) has broken.
I can't get one of these from TMC anymore -- the whole valve kit has to be bought for $120, and the bloody things were special-made for TMC.
Does anyone perchance have any of these things that they no longer need?
Thanks to all.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hi all, I need to have TEM screens recoated and heard that Grant Scientific is the place to contact. I called 803-799-6716 (a # I found in my address archives) and got the lovely fax machine scream sound. any help? thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I have examined amalgams in the SEM in the past. There is little problem with the mercury as it is amalgamated with silver. I have even used pure mercury in the SEM as a standard for WDS with no problem. I believe mercury is used in high vacuum diffusion pumps.
Good luck!
Sam O. Mancuso Group Leader, Physical Metallurgy Special Metals Corporation New Hartford NY 13413 phone: (315) 798-2920 fax: (315) 798-2001
I've got 2 really basic questions. First, I'm trying to determine the actual magnification power of my objectives. I'm using a calibration slide with 0.01 mm (I think that's the increments). What are the procedures for using this slide? I'm assuming I need to find out the diameter of the field and then do a reverse calculation. For example, if I have 36 lines (0.36 mm), using a 40x objective coupled with a 10x ocular, how do I calculate the actual mag power of my 40x obejctive. My second question is about fluorescent microscopy. I'm using a Zeiss fluorescence microscope with 40x and 63x Zeiss "neofluro" objectives, when I'm in focus, there are halos around the fluorescence image. What does this mean? Are my lens dirty? Oh, and one last thing. Does anyone know any good websites that teaches the basics about microscope maintenance and general troubleshooting faq?
I have a wonderful old Leitz Ortholux which I use to look at diatoms in my off time. Unfortunately, the coating on one of the prisms in the trinocular head has degraded with time, and shows up as a mottled shadow when viewing specimens.
Does anyone have recommendations and/or know of resources for reconditioning optics? Any suggestions concerning parts suppliers for old microscopes would be appreciated as well.
Thanks so much for your time!
Todd ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Todd A. Clason, Imaging Coordinator Department of Zoology University of Washington Box 351800 Seattle, WA 98195-1800
Grant Scientific Corp 1385 Rock Island Rd, Gilbert, SC 29054-8821 Phone: (803) 892-2841
Hope that's the right one.
Regards,
Rick Powell
} Hi all, } I need to have TEM screens recoated and heard that Grant Scientific is the } place to contact. I called 803-799-6716 (a # I found in my address } archives) and got the lovely fax machine scream sound. } any help? } thanks, } Beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } **************************************
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
Question: We recently bought a home microscopy kit and some "GUM MEDIA" was included in the set but was not explained in the manual. What is gum media and how would we use it? HELP!?
Osram Sylvania (http://www.sylvania.com) is a major manufacturer of tungsten wires. If you contact their local sales branch and request a small sample of the appropriate wire or range of wires, you'll get a lifetime supply. I've used their wire myself on XRF detectors and found them at least as good as the manufacturer's.
I can't think of many reasons why the whole detector should ever have to be replaced. You may need to clean it real well and change windows and wire. Any other parts that are damaged should be fairly easy to replace or duplicate.
On Monday, March 19, 2001 1:54 PM, "Edsworth-at-aol.com"-at-sparc5.microscopy.com [SMTP:"Edsworth-at-aol.com"-at-sparc5.microscopy.com] wrote: } } } I am in need of a gas flow proportional counter (GFPC) for a JEOL 733 } microprobe. Alternately, I may also need some W wire of the appropriate size } to restring an existing unit. Thanks for any help/advice. } } Ed Holdsworth } General Mgr. } SEMTEC Laboratories, Inc. } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
We have a liquid helium cold stage made by Oxford Instruments. It is used rarely because the safe handling precautions needed when using liquid helium (not to mention the cost) put many people off. However, if temperatures as low as 6K are a requirement in your experiment and the relevant safety precautions are taken, the thing works well. We also have Gatan's liquid nitrogen cold stage, which is much easier to use although only capable of minus 196C(?).
Hope this helps.
Kind regards,
Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
We have an Oxford Instruments double tilt He holder that fits both out JEOL 3000F and our 2010. We bought it about 2 years ago but Oxford Instruments have since been bought out by Gatan. We use the holder for a few days every couple of months and provided that you have common sense the hazards associated with handling liquid He are not a concern.
We are happy with the performance of the holder. Before buying we tested both the ultra-low temperature (5K) and low temperature (15K) versions and bought the 15K version as it looked more robust and we did not think that there was any real advantage to us to go to 5K. The holder we have gets below 10K.
It takes approximately 45 minutes to fill from room temperature and 30 minutes to refill when cold. If we operate at 20K it will hold the temperature for about 2 hours.
The tilt drives still work well at 10K and the image is stable enough to see gold fringes (0.2nm), after the cooling drift has stopped.
Since Gatan bought out Oxford Instruments I can only think of Fischione as a possible alternative supplier. It really is a shame that competition has reduced in the EM accessory field.
Good luck
Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
} Investigations of biomolecular interactions and mechanics (4 studentships) } These projects will employ SPM and state-of-the-art computational } methods to investigate the origins of biomolecular recognition and } protein folding or RNA folding pathways. Specifically, SPM will be } employed to characterize interaction forces between complementary } biological molecules e.g. individual coiled-coil proteins or } RNA-protein interactions (2 studentships). In parallel, two } studentships will aim to develop complementary computational methods } to further investigate the molecular origins of the obtained forces, } and also to explore the energetics of RNA/protein folding pathways. } } Ultra-high resolution studies of surface-chemistry and protein } structure (2 studentships) } In collaboration with Professor Richard James (Clinical Laboratory } Sciences), one project will utilize state-of-the-art SPM methods to } investigate, in-situ, the structure-function relationships of the } colicin class of bacterial membrane proteins. A second project will } aim to image foliate surfaces with high-resolution, and to } investigate the influence and surface distribution of various } agrochemicals (in collaboration with Syngenta-agrochemicals) } } The development and analysis of surface engineered biomimetic } systems for drug-delivery & tissue engineering (3 studentships) } In collaboration with the School's drug-delivery and tissue } engineering group, these projects will aim to develop novel } biomimetic polymeric surfaces for drug-delivery. In addition the } projects will employ state-of-the-art analytical techniques for } their surface-chemical characterization of polymer systems, } including SPM, contact angle, X-ray photoelectron microscopy and } time-of-flight secondary ion mass spectrometry. } } Single molecule optical microscopy using a single-molecule light } source (1 studentship) } This project will aim to address the current limitations of } near-field scanning optical microscopy, by replacing the physical } aperture by a small number of fluorescent molecule at the probe (as } proposed by W Heckl). The developed technology will be employed for } the high-resolution analysis of microfabricated biomolecular arrays. } } ________________________________________ } Phil Williams } Laboratory of Biophysics and Surface Analysis } School of Pharmaceutical Sciences and The Pharmacy School } University of Nottingham NG7 2RD } tel: +44 (0)115 9515025 } fax: +44 (0) 115 9515110 } http://www.nottingham.ac.uk/lbsa }
I am looking for quotes/sources for an anitivibration table for a TEM. The table will have to be larger than normal to accomodate some other instrumentation.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
I have imaged OLD amalgam without problem. FWIW, A copy of the image can be seen on my web site. No experience with "fresh" material.
http://woody.white.home.att.net
Woody -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi listers, } } Well, I stepped in it this time. I agreed to do some "gee-whiz" shots } for my dentist of a tooth with a rather large amalgam filling. Only } later } did it sink in that the mercury might be a problem under } vacuum and the } beam. Has anybody out there done this? I'd be looking at the } thing with } gold coating an a fairly light beam (say 10 kV), but } unfortunately, not } with } a cold stage. I've checked the archives and found one brave soul who } said } that he's done this on old amalgams. I can't really say how } old this one } is, but } I'm pretty sure it wasn't made yesterday. Call me paranoid, } but I'd like } some } more opinions before I do something really stupid! } } Cheers, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman } } }
Email: ken.andrew-at-bigpond.com.au Name: Ken ANDREW
Education: Graduate College
Location: Melbourne Australia
Question: Can I convert a Zeiss Jena polarized microscope from monocular to binocular. I have a student model from late-70's-early 80's which happens to also have metallographic attachment. E-bay often features binocular eyepieces. Is the barrel fitting common to all brands or just zeiss jena ? Does being a polarized scope make any difference ? I wear spectacles.
Hi there I am looking for a high resolution image of the MSA Logo in either Freehand, Illustrator or high resolution TIFF or JPEG, dows anyone have one or does anyone know who does have one? Many thanks.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
I would like to inform you that the new version of the homepage of the 5th Multinational Congress on ELectron Microscopy, that will be held in Lecce, Italy, from September 20-25 2001, is now ready and available at the URL: www.MCEM5.unile.it. In the "satellite events" section of the page, you will also find details about the "International School on Advances in Electron Microscopy in Materials Science" that will be held before the Congress.
The second circular/call for papers will be distributed to all who pre-registered very shortly.
Up to now we have 300 preregistrations from all over the world, and we really hope to have a nice and succesfull meeting.
Hope to see many of you in Lecce.
Best regards
Massimo
Dr. Massimo Catalano President of MCEM5 CNR-IME Via Arnesano 73100 Lecce - ITALY massimo.catalano-at-ime.le.cnr.it
Hi, I am looking for a new sputter coater to coat specimens for our FEG SEM, I was thinking of a Chromium coater but they are expensive and the films don¹t last very long (apparently). Is there a coater available that gives you small grain size and film longevity? Also are there brands who I should avoid for poor quality? Please reply off line as such opinions could be considered hurtful and libelous (even if preceded by IMHO!) ADVthanksANCE
-
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
I'd appreciate hearing from users of Disk Inspector software in the ISIS platform by Oxford. I'm especially interested in manual analysis of particles detected that are smaller than the minimum size established in the particle detection setup menu. Hope to share experiences and weed out some potential problems.
TIA
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
Hi Frank, Lets take your questions in reverse. It is likely that your 63x lens is an oil objective. The halo you see is most likely left over oil immersion media used on the lens. It is obligatory to use the oil for correct image formation. You indicate the 40x is presenting with the same symptoms, it doesn't necessarily have to be an oil objective however. It is possible it was designed as a dry lens but was accidentally smeared with oil. Specifications for each objective are on the body of the objective. oil lenses usually have a black colored indicator marking. Cleaning with non-abrasive lens paper (not chemwipes) and Windex wouldn't hurt either objective. You also need to make sure your light source, most likely mercury, is properly aligned. You may need help with this step, Zeiss manuals usually mention the steps required, but I strongly urge you seek out an experienced microscopist for this step. Actual magnification is a slightly tricky subject. Ocular x objective is a good range. It is further complicated by additional light gathering / magnification devices, i.e. field lenses, and the final form of the image. If you use 35mm film, print the micrograph, and measure your stage micrometer with a standard ruler to get a ratio which will indicate your final magnification. This figure will include any incidental magnification due to any other in line processes. If you are capturing video or digital, assign xx number for pixels to the stage micrometer. Good luck Ramin
-----Original Message----- } From: Frank Lee [mailto:flee-at-uhnres.utoronto.ca] Sent: Monday, March 19, 2001 6:55 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
I've got 2 really basic questions. First, I'm trying to determine the actual magnification power of my objectives. I'm using a calibration slide with 0.01 mm (I think that's the increments). What are the procedures for using this slide? I'm assuming I need to find out the diameter of the field and then do a reverse calculation. For example, if I have 36 lines (0.36 mm), using a 40x objective coupled with a 10x ocular, how do I calculate the actual mag power of my 40x obejctive. My second question is about fluorescent microscopy. I'm using a Zeiss fluorescence microscope with 40x and 63x Zeiss "neofluro" objectives, when I'm in focus, there are halos around the fluorescence image. What does this mean? Are my lens dirty? Oh, and one last thing. Does anyone know any good websites that teaches the basics about microscope maintenance and general troubleshooting faq?
At 11:28 PM 3/19/01 -0600, you wrote: } I've never heard of "Gum Media" can someone answer this question?
It must be "gum arabic", the water-soluble plant-based gum, for mounting specimens or even slide labels.
There are several mixtures that can be made with it. One comes to mind that involves chloral hydrate - but perhaps they don't let high school students work with that.
http://www.nmnh.si.edu/iz/copepod/techniques.htm has some recipes.
- John
} --------------------------------------------------------------------------- } } Email: JNDKepnerLovers-at-aol.com } Name: Josh Kepner } } Education: 9-12th Grade High School } } Location: City, State, Country } } Question: We recently bought a home microscopy kit and some "GUM MEDIA" was } included in the set but was not explained in the manual. What is gum media } and how would we use it? HELP!? } } ---------------------------------------------------------------------------
We are looking for a source for either an actual gold sputter target or plating service for our gold target for our Edwards 306A coater. Alternatively a source for gold foil would do. As usual time is of the essence. Vendors welcome. Thanks.
David O'Neil Institute for Marine Biosciences National Research Council 1411 Oxford St. Halifax , NS B3H 3Z1 Canada ph. 902-426-8258 fax. 902-426-9413 david.o'neil-at-nrc.ca
Mercury was indeed used in diffusion pumps, but as far as I know, that has been replaced by turbo pumps in most cases, especially due to the health hazards of mercury vapor.
As far as I know (and I am not a doctor!) mercury is fairly harmless in liquid form. It's the vapor that is dangerous. The problem is, that tiny drops of mercury tend to evaporate (due to surface tension), so you don't want to spill mercury. Keeping it in a bottle is not such a problem, as the vapor pressure is very low and it does not evaporate much.
Amalgam, however, is a different material. As Sam says, it's mercury amalgamated with silver. To find out what happens upon heating, you should probably consult some tables.
I heard, that the biggest health problem with Amalgam in teeth did not occur in patients, but in dentists. They had to handle the mercury and amalgamate it with silver all the time. Of course, there have been reports about health risks to patients also, which is probably why I haven't received any Amalgam fillings for a number of years (?).
If I had to look at a tooth with a filling, I would probably try to avoid heating it too much by keeping the beam current low.
Michael Bode
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: "SMancuso-at-specialmetals.com"-at-sparc5.microscopy.com [mailto:"SMancuso-at-specialmetals.com"-at-sparc5.microscopy.com] Sent: Monday, March 19, 2001 3:06 PM To: Microscopy Listserv
I have examined amalgams in the SEM in the past. There is little problem with the mercury as it is amalgamated with silver. I have even used pure mercury in the SEM as a standard for WDS with no problem. I believe mercury is used in high vacuum diffusion pumps.
Good luck!
Sam O. Mancuso Group Leader, Physical Metallurgy Special Metals Corporation New Hartford NY 13413 phone: (315) 798-2920 fax: (315) 798-2001
I need some ideas on how to reembed kidney tissue that has been embedded in Spurr's. I've heard of a method of using propylene oxide. Are there any other methods out there in TEM land???
Thanks in advance,
Donald G. Awbrey, HT(ASCP) QIHC Image Analysis / Electron Microscopy donaldawbrey-at-texashealth.org
It appears that Barnes and Noble (www.bn.com), Fatbrain (www.fatbrain.com) and Borders (www.borders.com) have it in stock. However, Amazon.com lists it as out of print.
Bookfinder.com is usually a good place to look for new and used books. However, for this book there were very few hits and it appears to list the author as Oliver Flint, but the ISBN number is the same as the book by Olga Flint.
Hope you find the book. I have it sitting on my desk now on interlibrary loan. I was wanting to look at it to determine if I would find it useful.
Judy Bowen Buckman Laboratories Memphis, TN USA
Previous message: I have been looking for a copy of "Food Microscopy", by Olga Flint. It is one volume of the Royal Microscopy Society handbook series. I saw copies available during the last MSA meeting, but cannot recall who the vendor was. Can somebody point me towards a company or individual who may have a copy of this book available for sale?
Karl Hagglund, Researcher P&G Food and Beverage Analytical and Microbiology 6071 Center Hill Ave., Box 117 Cincinnati, OH 45224 (513)634-0146
I have done extensive work on amalgams using SEM and TEM. There is no problem in the vacuum. Mercury is completely consumed/ reacted with alloy particles and the mercury containing phases are stable to 100C.
Tejpal Kaur Hooghan Agere Systems ---------- From: White, Woody N. [SMTP:nwwhite-at-mcdermott.com] Sent: Tuesday, March 20, 2001 8:24 AM To: 'James M. Ehrman'; 'Microscopy-at-MSA.Microscopy.Com' Subject: RE: dental amalgam in SEM?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
I have imaged OLD amalgam without problem. FWIW, A copy of the image can be seen on my web site. No experience with "fresh" material.
http://woody.white.home.att.net
Woody -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi listers, } } Well, I stepped in it this time. I agreed to do some "gee-whiz" shots } for my dentist of a tooth with a rather large amalgam filling. Only } later } did it sink in that the mercury might be a problem under } vacuum and the } beam. Has anybody out there done this? I'd be looking at the } thing with } gold coating an a fairly light beam (say 10 kV), but } unfortunately, not } with } a cold stage. I've checked the archives and found one brave soul who } said } that he's done this on old amalgams. I can't really say how } old this one } is, but } I'm pretty sure it wasn't made yesterday. Call me paranoid, } but I'd like } some } more opinions before I do something really stupid! } } Cheers, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman } } }
Hello, I'm having trouble finding a comprehensive source of info on what wavelengths to use with a multiphoton (Ti:sapphire) laser attached to a confocal to excite fluorochromes (if there is one yet...). Specifically, I'm having trouble with the alexa dyes, but a database of the usual and general dyes would help as well. I already have a cheat sheet from the confocal manufacturer listing the usual (FITC, TRITC, DAPI, etc..) and have "discovered" some not listed (Nile red, aniline blue, alexafluor 594).
Thanks! john shields university of georgia jshields-at-cb.uga.edu
I'm trying to prepare cryoTEM specimens of oil emulsions for one of my users, and it's clear that I don't have plunge-freezing, using ethane, figured out. I start with a Quantifoil grid, which I've glow-discharged, held at the edge with a pair of fine forceps. There's an o-ring keeping the forceps closed. I use a Pipetperson to place 4 microliters of oil emulsion on one side of the grid. I suspend the forceps/grid over a tiny cup of liquid ethane, centered within liquid nitrogen, using a guillotine apparatus with a foot-operated trigger. I blot the grid carefully with a piece of filter paper. At the instant in which the paper and grid separate, I trigger the guillotine. (The timing as I've described it differs from what I've read, but this is how I was shown to do it, and it seems to have worked -- once or twice. The people who showed me how to do this freeze virus suspensions, not oil emulsions.)
Now I have my forceps tip/grid in liquid ethane. I detach the forceps from the guillotine, keeping the grid suspended in the ethane, and then with a quick movement I immerse the grid in a small cup of liquid nitrogen that's suspended a few cm away in the same glass dewar. This is where I get a shallow dome of frozen ethane on the face of my grid. How do I prevent this from happening? Do I need to blot better, or longer, or blot and then leave the grid in air briefly before I dunk it? Or is there a trick to getting the ethane off at this point without damaging the grid?
Scott Robinson
Microscopy Suite Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana IL 61801 217 265-5071 (office); 217 244-6219 (fax) sjrobin-at-uiuc.edu
Awhile back I went to an art opening of Stefan Eberhard's photomicrographs. He's a researcher here at UGA at the Complex Carbohydrate Research Center. He does photomicroscopy using polarized light on commonplace chemicals (crystal images) such as cystine, niacin, vitamin C and my fav image Saccharin. I thought some of you might enjoy seeing the images. His website - http://home.att.net/~seberhard Something to do in your spare time;-) Beth
Disclaimer: I have no financial interest in this work. Just thought it was nice.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I'm trying to prepare cryoTEM specimens of oil emulsions for one of my users, and it's clear that I don't have plunge-freezing, using ethane, figured out. I start with a Quantifoil grid, which I've glow-discharged, held at the edge with a pair of fine forceps. There's an o-ring keeping the forceps closed. I use a Pipetperson to place 4 microliters of oil emulsion on one side of the grid. I suspend the forceps/grid over a tiny cup of liquid ethane, centered within liquid nitrogen, using a guillotine apparatus with a foot-operated trigger. I blot the grid carefully with a piece of filter paper. At the instant in which the paper and grid separate, I trigger the guillotine. (The timing as I've described it differs from what I've read, but this is how I was shown to do it, and it seems to have worked -- once or twice. The people who showed me how to do this freeze virus suspensions, not oil emulsions.)
Now I have my forceps tip/grid in liquid ethane. I detach the forceps from the guillotine, keeping the grid suspended in the ethane, and then with a quick movement I immerse the grid in a small cup of liquid nitrogen that's suspended a few cm away in the same glass dewar. This is where I get a shallow dome of frozen ethane on the face of my grid. How do I prevent this from happening? Do I need to blot better, or longer, or blot and then leave the grid in air briefly before I dunk it? Or is there a trick to getting the ethane off at this point without damaging the grid?
Dear Scott, Since the ethane is drawn up as you transfer the grid to the LN2, it would have to be removed during the time it is briefly in the N2 vapor (over the LN2 and ethane) before dunking into the small cup. This would expose your grid to higher temperatures and, possibly, water vapor, either of which will be far worse than the ethane. When the cryo-grid is inserted in the EM, the vacuum in the airlock should be good enough to cause the ethane to evaporate, so, unless the ethane affects the structure of the oil droplets, you should be better off just leaving the ethane in place. I would also doubt that there is a good cryogen which is hydrophyllic enough not to affect the oil, and is, at the same time, hydrophobic enough not to affect the water. If examination of the grids shows there to be a problem, you might try to withdraw the grid rapidly from the ethane--to try to draw up as little as possible--and move the grid quickly through a volume of dry N2 (the vapor over the LN2 should qualify) to try to dislodge or evaporate the ethane. I do not think that this procedure will be easy, and it could well raise the temperature of the specimen to unacceptable levels if the N2 vapor is not cold enough. Ideally you would want the temp to be just above the ethane boiling point. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Could this be gum arabic? It is a mounting medium for permanently mounting specimens under coverslips, and in another form (perhaps more dilute?) it can be used as a general adhesive.
} } Colleagues } } I've never heard of "Gum Media" can someone answer this question? } } Nestor } } --------------------------------------------------------------------------- } } Email: JNDKepnerLovers-at-aol.com } Name: Josh Kepner } } Education: 9-12th Grade High School } } Location: City, State, Country } } Question: We recently bought a home microscopy kit and some "GUM MEDIA" was } included in the set but was not explained in the manual. What is gum media } and how would we use it? HELP!? } } ---------------------------------------------------------------------------
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Scott Robinson asked: ..I get a shallow dome of frozen ethane on the face of my grid. How do I prevent this from happening?
The answer depends upon how much ethane is retained and where it is located. A small covering of solid ethane actually serves as an additional frost shield. This will pump off in the airlock of the microscope. On our Philips CM-20 we actually insert the holder into the airlock and pump for a while before dumping the LN2 and inserting the holder. This will work on a standard airlock with increased pumping time by the cryo software. Our airlock is a special turbo-pumped system that presumably gives us a cleaner vacuum.
The problem comes when there is so much ethane film that it interferes with loading in the holder. When a solid ethane film splits off the grid, it usually takes the sample with it. This is very annoying (to say the least). We have blotted the grid on a dry piece of filter paper in the cryochamber prior to plunging in the liquid nitrogen. This works ok if you work fast. It is another opportunity to get frost contamination on the sample so we only do this as a last resort...
I'd be interested in hearing what problems you have with oil emulsions. Our work, along with that of Ishi Talmon, suggests when you have high concentration of oil phase in contact with ice (vitreous or otherwise) that radiation damage (visible as bubbling) is worse than in most aqueous based systems.
Best Regards,
John Minter Eastman Kodak Co. Analytical Technology Division Rochester, NY 14650-2152 Phone: (716) 722-3407 FAX: (716) 477-9303
I'm passing on a question from a colleague. Does anyone know of a technique for etching LR White from sections on a slide? I gave him all the info I had on epoxy removal using ethoxide and so on, but I've been unable to locate any references specific to LR White.
Thanks. Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
This course has been designed to teach microscopists the preparation of high quality SEM and Light Microscopy specimens using new broad ion beam techniques and instrumentation. The two-day practical course intended to offer end-users an alternative/replacement method to the traditional "wet" chemical etching technique, used as final step in the sample preparation process.
Please visit http://www.gatan.com for details.
To register, please e-mail info-at-gatan.com and request a Gatan Schools Registration Kit.
Olympus POS for sale. Good monocular PLM student scope. Comes with Mahogany case, 10x, 20x and 40x stage, Leitz spindle. All offers welcomed. Lou Solebello microls1297-at-mindspring.com
Dear Scott, Why don't you try pure grade propane? The freezing procedure is exactly the same as with the ethane. You will not get frozen ethane, but may have contamination from comercial propane if you do not find pure one. svetla
Scott Robinson asked: .. Do I need to blot better, or longer, or blot and then leave the grid in air briefly before I dunk it? Or is there a trick to getting the ethane off at this point without damaging the grid?
You should use a small cryochamber with enough space for the ethane cup, the specimen holder, some small tools and a piece of filter paper all located above a boiling liquid nitrogen bath. The evaporating nitrogen prevents all parts from contamination with ice. After immersing the grid into the ethane the grid should be blotted on the dry filter paper where the excess of liquid ethane is drained off. When the specimen holder is located near by the transfer from liquid ethane to the holder via the filter paper can be done very fast. In the humidity free atmosphere of the chamber the intermediate storage in liquid nitrogen can be omitted.
Best regards, Markus
******************************************* Dr. Markus Drechsler Friedrich-Schiller-Universitaet Jena Institut fuer Pharmazie Lehrstuhl fuer Pharmazeutische Technologie Lessingstr.8 D-07743 Jena
You can order 'Food Microscopy', and indeed any other Royal Microscopical Society Handbook, from the publisher, Bios Scientific.
Contact sales-at-bios.co.uk to order directly, or visit www.bios.co.uk or our website, www.rms.org.uk/handbook.html if you want more information on any of the RMS handbooks.
Best wishes
Sue ************************************************************************** Sue Betteridge, Publications Officer, Royal Microscopical Society 37/38 St Clements, Oxford OX4 1AJ, UK. Tel +44 (0)1865 248768, fax +44 (0)1865 791237, email jmicrosc-at-rms.org.uk, website http://www.rms.org.uk **************************************************************************
My sugestion would be to either build or buy a carbon evaporator. While it may not be impossible to sputter carbon with more exotic sputtering devices, a Hummer V will not work.
Wooody White ------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------. } } } I have a Technics sputter coater- Hummer -V. I'd like help finding a } carbon source for it. } Any ideas? } Thank you. } } }
This is a discussion archived at "Tips & Tricks" There might be more than one, but this is the first I hit with the search.
The rest of the archive can be found at:
www.biotech.ufl.edu/~emcl
Follow the Tips & Tricks link.
I will begin adding material to the site soon. Anything you have found useful in the course of your studies would be welcome. Simply forward it to whittaker.scott-at-nmnh.si.edu. You will of course be given the credit.
Scott Whittaker SEM Lab Manager National Museum of Natural History Smithsonian Institution 10th & Constitution Ave, NW Washington DC 20560-0104 202-357-1651 {*^-at-)~~~ It was recently discovered that research causes cancer in rats. ~~~(-at-^*}
} } } "Tindall, Randy D." {TindallR-at-missouri.edu} 03/20/01 05:45PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I'm passing on a question from a colleague. Does anyone know of a technique for etching LR White from sections on a slide? I gave him all the info I had on epoxy removal using ethoxide and so on, but I've been unable to locate any references specific to LR White.
Thanks. Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I would like to hearing from you opinions about Scanning microscopes. We have a limited fund to buy a new scanning electron microscope and we have just received a proposal from a Japanese company named SHIMADZU. They offered us a scanning electron microscope model SS-550. I confess that I don't know much about this company and neither about the quality of its instruments (particularly SEM). I would appreciate if someone out there could give me an opinion about this equipment. Right now we can buy a unit without the EDS analitical system which we are planning to buy later. Then I also would like to know if we would buy only the image unit first we could buy later the EDS system (from the same company) separately to upgrade the same model (SS-550). The vendor said that it is possible, but sometimes it is hard to trust in sellers especially when you live in South America.
Thank all of you
Best wishes,
Leonardo -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
I am trying to find a procedure to detect cellular and membrane cholesterol at the EM level in RBC's. Using the standard EM processing techniques, cholesterol is dissolved out leaving familiar clefts. Does anyone out there know of a protocol where I can preserve the cholesterol at the EM level and still have decent ultrastructure.
Howard Mulhern Children's Hospital Dept. of Pathology Surgical Path EM Facility Boston, 02115, Ma.
I'm looking for a simple DLS program, freeware and preferably source code. This would be "distance least squares" for cleaning up atomic positions (for example those input by hand from a model), minimizing the mean squared deviation from expected bond angles and spacings.
I'm particularly interested in code which would do this for silicates (SiO2), using typical ~1.6 Angstrom Si-O distance and 109 degree Si-O-Si angle.
I actually found something of this kind, ostensibly free for downloading, at
http://www.kristall.ethz.ch/LFK/software/xrs/
However, I'm consistently refused the ftp connection listed there, and have gotten no response to queries.
Does anyone know of other sources of such software?
Greetings, If you look up protocols for doing PAS staining to show polysaccharides (in plant and I suppose any tissue), they start with an aldehyde quench, with either of two chemicals. Years ago we discovered that for our purposes we don't need that step, and now I have forgotten the name of the chemicals. The names are something like DNPH and Dimedone but I can't remember exactly. In any book of botanical microtechnique that gives the PAS procedure, they should be there.
Sorry about my flawed memory.
Tobias
} } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller
Bodycote Ortech is seeking an experienced electron microscopist for our physical characterization group. The successful candidate will provide hands-on microscopy services to our clients in the healthcare, general manufacturing and materials sectors. You will characterize a wide variety of man-made and naturally occurring materials, and investigate materials breakdown and failure in support of manufacturing processes. You will work with our clients to define project scope, write quotations, perform analytical work and prepare project reports.
Your expertise lies in analytical scanning electron microscopy, including low temperature SEM. You are adept at using light microscopy and image analysis techniques. You enjoy problem solving and research activities, manage your time well and are able to work effectively in a team atmosphere. Ideally you have a graduate degree in chemistry, biology or a related field and more than three years experience working in an electron microscopy laboratory.
To join our team, please forward your resume to Human Resources Department, Bodycote Ortech, 2395 Speakman Drive, Mississauga, Ontario, L5K 1B3, email bruce.a-at-bodycote.ca Fax: (905) 823-1446
Ammonium chloride (50-100 mM) in PBS or glycine (sorry, don't remember concerntration--probably 1 or 2 % ??) in PBS have been reported. We use NH4Cl.
Sara Miller
On Wed, 21 Mar 2001, Caroline Miller wrote:
} Date: Wed, 21 Mar 2001 12:24:55 -0600 } From: Caroline Miller {camiller-at-creighton.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Aldehyde Removal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I'm afraid that I have never heard of Shimadzu SEM's or EDS's. Moneywise, I evaluated JEOL, Hitachi, and KLA/Tencor (Amray) SEM's and found that JEOL was by far the most economical for equivalent systems. (We also purchased an EDAX EDS system to go with it.)
I have no interest in JEOL or EDAX (other than being a happy customer.)
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
Here are the replies I have received so far on my question about sputter coaters. They are as I have received them except I have removed the senders names. Some are obviously from vendors. These are NOT my opinions I am simply reflecting what I have learned. YMMV.
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Dear Sir, I saw your posting on the MSA list server and wanted to know if you were aware of Cressington products. You can view our web site at www.cressington.com {http://www.cressington.com} . Also I would offer to come to your lab to demonstrate our 208HR sputter coater. As far as price being an issue we are always tailoring quotes around tight budgets. Please feel free to contact me if you would like more information.
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We have a KMC Ion-Tech, which I've been told has been taken over by South Bay Technologies. This is good. KMC was everything bad in a company. If South Bay has redesigned -- extensively -- the ion-coater, then this might be a good machine. Otherwise, I'd avoid it like the plague. Even though I generally think good thoughts about South Bay.
When the thing works right, we use it for platinum-coating specimens for FESEM down to 1 or 2 nm (measured to 0.7 nm, but that really requires the right specimen). The Pt coat is very fine. I saw grain structure once at something over 200,000X (might have been 400,000, this was a while ago), but typically I see no structure.
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We used a sputter coater with a chromium target for awhile, with good results. You are correct, however, in that the chromium begins oxidizing immediately and will quickly form an insulating oxide layer. Samples should be viewed as quickly as possible after coating and stored in the best vacuum possible if repeated viewings are required. Chromium is not a good way to go for specimens that need to be kept for repeated viewings.
Our compromise was to purchase a platinum target, which provides somewhat larger grain size than chromium, but smaller than gold or gold-palladium. Platinum, of course, does not oxidize. Platinum and other targets are available at very reasonable cost (relative to the sputter coaters' manufacturer's prices) from Abe Dayani at Refining Systems, Inc. He can be reached at P.O. Box 72466, Las Vegas, NV 89170. Phone is (702) 368-0579 and fax is (702) 368-0933. I have dealt with him before and found him to be very helpful and knowledgeable.
We are using an EMITECH sputter coater currently, but I cannot in good faith recommend them at this time. The coater works well for the most part, but the company is recovering from some internal problems and their service is spotty (to be kind). I think that if or when the company is over its present difficulties that this will again be a wonderful line of equipment.
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I can't address relative grain size and longevity, nor Cr coats specifically, but the carbon coater we opted for last year is the Emitech 950C. It also has an easily interchangeable metal head. We felt this turbo unit was the best value by a considerable stretch. So far we've not done metal [though may do some Au-Pd fairly soon], only carbon, but the carbon coats have been very consistent. URL:
http://www.emitech.demon.co.uk/K950.htm
US contact: Linda Dailey {emitech-at-earthlink.net}
If you feed power to your roughing pump through the K950, allowing automatic control of the rotary by the coater, there is a fuse/circuit issue we had that is solved by using the auto control for a relay switch.
Best of luck.
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The 681 and the 682 BOTH provide an amorphous coating. The 682 also does ecthing. The 681 is coating ONLY.
If you want to talk with someone about applications, please call Dick Mitro at 925-224-7319.
If you don't mind seeing the coating(s), then a cheap $5-$10K coater will suffice.
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I recently went through this decision process for a HR coater for our new LEO 1550. I decided on the Cressington 208HR. It can do Cr, but also Au or Pd or Pt or Au/Pd. It has several nice features such as a rotary/planetary/tilting stage to allow for a thin 2nm coating even on rougher samples. My research led me to this coater or one by Emitech. I think both are good coaters and both were in the $25,000 range. We are taking delivery of the Cressington in about X weeks. If you are still looking at that point I can tell you what I think once I get a chance to use it on several of our samples. Of course I had them coat some samples for us before I made my decision but lets see how it works in the lab. We run a wide range of samples, polymer, metal, ceramic. Good luck.
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I saw this posted on the listserver and thought you might have an interest in our IBS/e Ion Beam Sputter Deposition and Etching System. It is on the pricey side compared to a sputter coater ($50-60k), but can deposit very thin, uniform, small grain films using chromium, iridium, carbon etc. It has many advantages over magnetron systems.
If it is of interest, I would be happy to send you more detailed information. Please let me know.
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IMHO! I know it is not the least expensive choice, but I recently purchaced an SBT IBSS (Ion Beam Sputter/etch System). It does a fantastic job with Ir. We have a LEO 1530 (FEG) and routinely image up to } x200,000 with no evidence of the coating structure. It's durability is fine for us, we have specimens in us often for several days and up to a week or more, without significant deterioration in conductivity. Vince Carlino is with them (South Bay) now and this coater is basically his previous system in a new and improved system/package. Of course you can use Cr or Pt or virtually any other metal to "coat", but we have had tremendous success with Ir. It is a very flexible, durable, and effective instrument for us.
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For DC sputtering the next best option is Pt, better than Au or Pd or Au/Pd mix.
However Cr is best but has all of the inherent issues that you have mentioned, We are currently evaluating using Irridium for DC sputtering, all the advantages of Cr in grain size but none oxidizing.
However very hard to obtaining targets of sufficient purity and very expensive.
Have a look at our site in the US this shows our equipment and has some tech information that you may find useful.
www.empdirect.com
The Emitech K575X turbo sputter coater is designed with Cr and FE sem in mind, the unit may also be used to coat with noble metals as mentioned above.
Please call or email if you have any questions or would like price information on any of our products.
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I too am interested in purchasing another sputter coater for use with our JEOL FEG-SEM. For the last 8 years or so we have been using a Plasma Sciences CrC-100 coater with a chromium target. Generally the unit has served us well, however, I have had two nagging concerns with respect to this coater, 1) its age - it's becoming tempermental of late and 2) Plasma Sciences is no longer in business, service and parts have been taken over by Torr International. I don't know where or from whom you were informed regarding problems associated with chromium. We have not had that experience.
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Denton desk II did a good job while I was at university of xxxx with just gold/palladium on a hitachi s-4000 FESEM.
I now have a Cressington Scientific 108 SE automatic but it is so new I have yet to really put it through the paces. They say it does a wonderful job and for the price it better. It is also what the FEI applications lab has for the FEG demonstrations. Those pics were impressive
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Take a look at URL http://www.2spi.com/catalog/osmi-coat.html
It is now almost impossible to sell a chromium coater in Japan; the Japanese learned over the past few years that nothing comes even close to what the osmium coater will do. I know that sounds almost too good to be true but consider a) the metal layer is completely amorphous, there is absoulutely no grain size, and of course, with Cr, there is some point where you do start to see the grain, and b) the metallization has the same inertness as gold or Pt, so unlike Cr which starts to oxidize almost immediately, the osmium metal coating lasts almost forever.
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I've used gold-palladium targets with good results in ordinary sputter coaters. Emitech and Anatech/Hummer are two brands we've had good luck with.
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Pertaining to your recent question posted to Microprober list, perhaps you may want to try Polaron sputter coater SC series at reasonable price, I have been using few brands of sputter coater for FESEM, I think this is recommended.
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Would appreciate hearing what you learn about sputter coaters.
Our 15-year-old Au-coater (Denton) has been reliable and user-friendly, but considering its age, I could be faced with replacing it before too much longer.
Topographic artifacts produced by the Au-sputter have been of interest because of the 'nannobacteria' problem. With our little garden-variety W-filament SEM, a 30 sec coating does not produce detectable artifacts (below about 50,000x) whereas longer coating times may (See Fig 7, p. 588 in Folk and Lynch, JSR v. 67).
In the FE-SEM, samples coated with our sputter for 45 sec begin to show potentially troubling textures (artifacts?) around 80,000x.
Thanks for any info you can share!
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I purchased a Cressington 208 HR Sputter Coater for use with our FEG SEM. It has worked very nicely. I decided that the Cr target was too much of a hassle for routine prep work, so for the majority of work, I use Au/Pd and apply a coating of from 1 to 8 nm in thickness, depending on my samples. The system has been very reliable, pumps down quickly, and releases the vacuum quickly. The little shield that swings in and out works to protect the sample from the initial oxidized Cr from the target surface, but when the oxide layer is sputtered off, you swing the small shield out of the way to coat the sample with Cr. The rotating, tilting stage allows for good coverage of the sample with the target material. I know there are other good brands out there, but the Cressington works very well.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
SHIMADZU is definitely a small player here in the US.
In the 27 yeatrs I have been involved in SEMs, I have never heard of them.
Regards,
Earl
----- Original Message ----- } From: "Leonardo Lagoeiro" {lagoeiro-at-degeo.ufop.br} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 21, 2001 6:57 AM
I didn't know that Shimadzu (who are a large and reputable scientific equipment manufacturer who afew years ago bought Kratos)) made an SEM, and a search of their website www.shimadzu.com revealed none. However, I see that their regional Brasil website lists not only the SEM 550, but also the EPMA 1600! I remember hearing, years ago, that they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that may not have been so.
I'd be very interested to hear more about either instrument.
Anybody know anything?
cheers
rtch
} } Hi there, } } I would like to hearing from you opinions about Scanning } microscopes. We have a limited fund to buy a new scanning electron } microscope and we have just received a proposal from a Japanese } company named SHIMADZU. They offered us a scanning electron } microscope model SS-550. I confess that I don't know much about this } company and neither about the quality of its instruments } (particularly SEM). I would appreciate if someone out there could } give me an opinion about this equipment. Right now we can buy a unit } without the EDS analitical system which we are planning to buy } later. Then I also would like to know if we would buy only the image } unit first we could buy later the EDS system (from the same company) } separately to upgrade the same model (SS-550). The vendor said that } it is possible, but sometimes it is hard to trust in sellers } especially when you live in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } ---
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
G'day Caroline, You need to carry out an "aldehyde blockade" process, This will convert all aldehyde groups. Including tissue aldehyde groups. Method: A saturated solution of 2,4-DNP(2,4dinitrophenyl hydrazine) in 15% acetic acid. Time 0.5Hrs. Wash thoroughly in tap water. Time 0.5Hrs
If you are going on to do a PAS reaction you then begin your oxidation step with 1% Periodic acid. Regards JVN
Caroline Miller wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
Applied Molecular Evolution (AMEV), is an early stage biotech company with secure financial funding and high growth potential. We focus on novel high throughput approaches to drug discovery, screening compound libraries against thousands of variants of G protein coupled receptors using highly advanced instrumentation.
We are looking for an entry level Scientist in the department of Fluorescence Imaging. The candidate is expected to have a Ph.D. and be experienced in fluorescence imaging. Experience with Ca2+-sensitive dyes and G Protein coupled receptors is a plus.
We are willing to recognize the added value of highly skilled and motivated individuals. Novasite employees receive excellent benefits and compensation, including equity participation and generous performance-linked incentives.
Please Reference Job Code: NR0321A jobs-at-novasite.com Novasite Pharmaceuticals 3520 Dunhill Street San Diego CA 92121 Fax: (858) 597-4950
San Diego based Novasite Pharmaceuticals, Inc., a subsidiary of Applied Molecular Evolution (AMEV), is an early stage biotech company with secure financial funding and high growth potential. We focus on novel high throughput approaches to drug discovery, screening compound libraries against thousands of variants of G protein coupled receptors using highly advanced instrumentation.
We are looking for an entry level Scientist in the department of Fluorescence Imaging. The candidate is expected to have a Ph.D. and be experienced in fluorescence imaging. Experience with Ca2+-sensitive dyes and G Protein coupled receptors is a plus.
We are willing to recognize the added value of highly skilled and motivated individuals. Novasite employees receive excellent benefits and compensation, including equity participation and generous performance-linked incentives.
Please Reference Job Code: NR0321A jobs-at-novasite.com Novasite Pharmaceuticals 3520 Dunhill Street San Diego CA 92121 Fax: (858) 597-4950
We are preparing to embark on an in-depth project with outside funding that will require, among other things, 3-D reconstruction of series' of transmission electron micrographs of sections of biological material. We don't have a great deal of recent experience in this area and would welcome (off list if you are a commercial concern) suggestions for the "best" or most capable, or most flexible or useful 3-D reconstruction software including what computer platform is required and what configuration is considered adequate or better for this type of work. In addition, what digital data collection system currently available would be best for this type of work? We presently have a Philips CM12S STEM with a slow scan CCD camera with a 1k x 1k chip that has done good service for an extended period, but it seems that it might be a bit difficult to use for serial section recording and image orientation in preparation for 3-D reconstruction. If something new in a hardware-software package is available, we would be interested to hear how it works for others, or to hear directly from vendors off list.
Separately, it is possible that TEM tomography will also be in our future - It is our understanding that either at least IVEM or an energy filtered TEM are required to do useful tomography of thicker sections (0.25 - 0.5 micron thick). What are others in this area of interest using for microscopes, computers, and software? Has something of a "standard" come about, or are people still writing their own code and putting systems together from scratch?
Thanks in advance for any and all information and advice.
Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
I do not know this company either, but why do you have proposal just from one company? Ask other vendors and compare their proposals. As for EDS it is better to by system directly from manufacturer and ask them in advance proposals too - if SEM you choose is not in their "standard microscopes" list they may charge you extra for an adapter.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Leonardo Lagoeiro [mailto:lagoeiro-at-degeo.ufop.br] } Sent: Wednesday, March 21, 2001 8:57 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM inquiry } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi there, } } I would like to hearing from you opinions about Scanning microscopes. } We have a limited fund to buy a new scanning electron microscope and } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). I } would appreciate if someone out there could give me an opinion about } this equipment. Right now we can buy a unit without the EDS } analitical system which we are planning to buy later. Then I also } would like to know if we would buy only the image unit first we could } buy later the EDS system (from the same company) separately to } upgrade the same model (SS-550). The vendor said that it is possible, } but sometimes it is hard to trust in sellers especially when you live } in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br }
Caroline didn't say whether she wanted the aldehyde procedure for LM or EM, but I suspect the 15% acid wouldn't be to good for ultrastructural studies.
On Thu, 22 Mar 2001, John V Nailon wrote:
} Date: Thu, 22 Mar 2001 09:14:34 +1000 } From: John V Nailon {J.Nailon-at-mailbox.uq.edu.au} } To: Caroline Miller {camiller-at-creighton.edu} } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: Aldehyde Removal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } G'day Caroline, } You need to carry out an "aldehyde blockade" process, This will convert } all aldehyde groups. Including tissue aldehyde groups. } Method: A saturated solution of 2,4-DNP(2,4dinitrophenyl hydrazine) in } 15% acetic acid. Time 0.5Hrs. } Wash thoroughly in tap water. Time 0.5Hrs } } If you are going on to do a PAS reaction you then begin your oxidation } step with 1% Periodic acid. } Regards } JVN } } Caroline Miller wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I'm looking for a method to remove aldehydes from tissue before } } processing? Thanks, Caroline Miller } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Could it be that Shimadzu are now the manufactures/distributors of the Topcon/Akashi/ISI range of instruments?? Regards JVN
Ritchie Sims wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I didn't know that Shimadzu (who are a large and reputable scientific } equipment manufacturer who afew years ago bought Kratos)) made an } SEM, and a search of their website www.shimadzu.com revealed none. } However, I see that their regional Brasil website lists not only the } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } may not have been so. } } I'd be very interested to hear more about either instrument. } } Anybody know anything? } } cheers } } rtch } } } } } Hi there, } } } } I would like to hearing from you opinions about Scanning } } microscopes. We have a limited fund to buy a new scanning electron } } microscope and we have just received a proposal from a Japanese } } company named SHIMADZU. They offered us a scanning electron } } microscope model SS-550. I confess that I don't know much about this } } company and neither about the quality of its instruments } } (particularly SEM). I would appreciate if someone out there could } } give me an opinion about this equipment. Right now we can buy a unit } } without the EDS analitical system which we are planning to buy } } later. Then I also would like to know if we would buy only the image } } unit first we could buy later the EDS system (from the same company) } } separately to upgrade the same model (SS-550). The vendor said that } } it is possible, but sometimes it is hard to trust in sellers } } especially when you live in South America. } } } } Thank all of you } } } } Best wishes, } } } } } } Leonardo } } -- } } --- } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
I was searched Shimadzu web site at http://www.shimadzu.com, but there is no
any information about scanning electron microscope.
Henrik
Ritchie Sims wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I didn't know that Shimadzu (who are a large and reputable scientific } equipment manufacturer who afew years ago bought Kratos)) made an } SEM, and a search of their website www.shimadzu.com revealed none. } However, I see that their regional Brasil website lists not only the } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } may not have been so. } } I'd be very interested to hear more about either instrument. } } Anybody know anything? } } cheers } } rtch } } } } } Hi there, } } } } I would like to hearing from you opinions about Scanning } } microscopes. We have a limited fund to buy a new scanning electron } } microscope and we have just received a proposal from a Japanese } } company named SHIMADZU. They offered us a scanning electron } } microscope model SS-550. I confess that I don't know much about this } } company and neither about the quality of its instruments } } (particularly SEM). I would appreciate if someone out there could } } give me an opinion about this equipment. Right now we can buy a unit } } without the EDS analitical system which we are planning to buy } } later. Then I also would like to know if we would buy only the image } } unit first we could buy later the EDS system (from the same company) } } separately to upgrade the same model (SS-550). The vendor said that } } it is possible, but sometimes it is hard to trust in sellers } } especially when you live in South America. } } } } Thank all of you } } } } Best wishes, } } } } } } Leonardo } } -- } } --- } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
-- Henrik Kaker, Ph.D. Metal Ravne d.o.o. SEM-EDS Lab Koroska cesta 14, 2390 Ravne Slovenia Phone: +386 02 82 21 131 Fax: +386 02 82 20 436 http://www.kaker.com
Some questions arising: 1) The use of iridium for sputtering is news to me, and looks interesting. I would be interested to see a comparison of iridium film structure with platinum etc. Can a garden variety gold or gold/palladium unit sputter iridium, or are more exotic conditions required?
2) Does anyone have experience of the practical and safety issues arising from osmium coaters? Is the release of osmium tetroxide vapour to room atmosphere well controlled? How economical are they to run? Would they be practical in an environment where there could be several days or even weeks between coating runs? Presumably there are no safety issues with the coated specimens if the metal is inert. True?
Chris
snip
IMHO! I know it is not the least expensive choice, but I recently purchaced an SBT IBSS (Ion Beam Sputter/etch System). It does a fantastic job with Ir. We have a LEO 1530 (FEG) and routinely image up to } x200,000 with no evidence of the coating structure. It's durability is fine for us, we have specimens in us often for several days and up to a week or more, without significant deterioration in conductivity. Vince Carlino is with them (South Bay) now and this coater is basically his previous system in a new and improved system/package. Of course you can use Cr or Pt or virtually any other metal to "coat", but we have had tremendous success with Ir. It is a very flexible, durable, and effective instrument for us.
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For DC sputtering the next best option is Pt, better than Au or Pd or Au/Pd mix.
However Cr is best but has all of the inherent issues that you have mentioned, We are currently evaluating using Irridium for DC sputtering, all the advantages of Cr in grain size but none oxidizing.
However very hard to obtaining targets of sufficient purity and very expensive.
Have a look at our site in the US this shows our equipment and has some tech information that you may find useful.
www.empdirect.com
The Emitech K575X turbo sputter coater is designed with Cr and FE sem in mind, the unit may also be used to coat with noble metals as mentioned above.
Please call or email if you have any questions or would like price information on any of our products.
___________________
I too am interested in purchasing another sputter coater for use with our JEOL FEG-SEM. For the last 8 years or so we have been using a Plasma Sciences CrC-100 coater with a chromium target. Generally the unit has served us well, however, I have had two nagging concerns with respect to this coater, 1) its age - it's becoming tempermental of late and 2) Plasma Sciences is no longer in business, service and parts have been taken over by Torr International. I don't know where or from whom you were informed regarding problems associated with chromium. We have not had that experience.
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Denton desk II did a good job while I was at university of xxxx with just gold/palladium on a hitachi s-4000 FESEM.
I now have a Cressington Scientific 108 SE automatic but it is so new I have yet to really put it through the paces. They say it does a wonderful job and for the price it better. It is also what the FEI applications lab has for the FEG demonstrations. Those pics were impressive
___________________
Take a look at URL http://www.2spi.com/catalog/osmi-coat.html
It is now almost impossible to sell a chromium coater in Japan; the Japanese learned over the past few years that nothing comes even close to what the osmium coater will do. I know that sounds almost too good to be true but consider a) the metal layer is completely amorphous, there is absoulutely no grain size, and of course, with Cr, there is some point where you do start to see the grain, and b) the metallization has the same inertness as gold or Pt, so unlike Cr which starts to oxidize almost immediately, the osmium metal coating lasts almost forever.
___________________
I've used gold-palladium targets with good results in ordinary sputter coaters. Emitech and Anatech/Hummer are two brands we've had good luck with.
___________________
Pertaining to your recent question posted to Microprober list, perhaps you may want to try Polaron sputter coater SC series at reasonable price, I have been using few brands of sputter coater for FESEM, I think this is recommended.
___________________
Would appreciate hearing what you learn about sputter coaters.
Our 15-year-old Au-coater (Denton) has been reliable and user-friendly, but considering its age, I could be faced with replacing it before too much longer.
Topographic artifacts produced by the Au-sputter have been of interest because of the 'nannobacteria' problem. With our little garden-variety W-filament SEM, a 30 sec coating does not produce detectable artifacts (below about 50,000x) whereas longer coating times may (See Fig 7, p. 588 in Folk and Lynch, JSR v. 67).
In the FE-SEM, samples coated with our sputter for 45 sec begin to show potentially troubling textures (artifacts?) around 80,000x.
Thanks for any info you can share!
___________________
I purchased a Cressington 208 HR Sputter Coater for use with our FEG SEM. It has worked very nicely. I decided that the Cr target was too much of a hassle for routine prep work, so for the majority of work, I use Au/Pd and apply a coating of from 1 to 8 nm in thickness, depending on my samples. The system has been very reliable, pumps down quickly, and releases the vacuum quickly. The little shield that swings in and out works to protect the sample from the initial oxidized Cr from the target surface, but when the oxide layer is sputtered off, you swing the small shield out of the way to coat the sample with Cr. The rotating, tilting stage allows for good coverage of the sample with the target material. I know there are other good brands out there, but the Cressington works very well.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
As is mentioned by other members you are unable to use a carbon target with your coater but you may possibly be able to obtain a "carbon string" head and power pack for it?
This package usually contains a new top plate with through terminals, a power pack (not much different to adding a car battery) and some safety interconnections. Most sputter coater manufacturers have this type of accessory.
You are basically using a carbon fibre which you "burn" at high temperature to evaporate the carbon. The coating is not from a point source (as would the conventional carbon electrodes used in a high vacuum coater) so it is better for SEM samples. However the carbon is very soft and easily damaged even when evaporated under (the desired) best sputter coating vacuum level.
In short it is a quick and easy SEM technique but it does not provide a coat as efficient as a sputtered metal, or provide a coating that would allow the production of free floating carbon films here, a high vacuum coater is essential .
Good luck but talk to the manufacturer.
Steve Chapman Senior Consultant Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I have undertaken several SEM tests with EDS analysis of minute "Black Spots" that we are having problems with in the Ultra High Molecular Weight polyethylene components that we manufacture at our plant. We thought that identifying the spots would help us eliminate the problem. To a certain extent it did. i.e. any Fe,Cr& Ni that was found together is almost certainly stainless steel splinter.
However, it is the other results that are proving difficult to identify. Fe is present along with S, Si & Na? Another spot yielded Si, K ,Na and Ca together. A further example of a spot would be; Fe,Ca,Si,Ti,K and Na.
Would anyone have any ideas on how to identify what these particles are?
Any information or help would be gratefully appreciated.
It's difficult to ID material without knowing how the spectra were collected and what elements were dominant. You list Si in the three spots that you describe. If it was the major foreign element, then you might have some silicate material.
If available it would be good to try Micro IR to further characterize the spots.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027
paulfinnegan-at-ireland.com on 03/22/2001 07:27:09 AM To: Microscopy-at-sparc5.Microscopy.Com cc:
Hi all,
I have undertaken several SEM tests with EDS analysis of minute "Black Spots" that we are having problems with in the Ultra High Molecular Weight polyethylene components that we manufacture at our plant. We thought that identifying the spots would help us eliminate the problem. To a certain extent it did. i.e. any Fe,Cr& Ni that was found together is almost certainly stainless steel splinter.
However, it is the other results that are proving difficult to identify. Fe is present along with S, Si & Na? Another spot yielded Si, K ,Na and Ca together. A further example of a spot would be; Fe,Ca,Si,Ti,K and Na.
Would anyone have any ideas on how to identify what these particles are?
Any information or help would be gratefully appreciated.
There exists a homepage of 3-Dimensional Microscopy Labs (http://3dem.sdsc.edu/) in which are listed various packages for 3D-EM Image Analysis. Additionally a mailing list exists.
BTW: A nice animation can be found in a Web-Page of the University of Utrecht (The Netherlands): http://emsaserv.bio.uu.nl/3dem/ANIMATED_INTRODUCTION/animated_introduction_1.html
I hope this helps a little bit. Of course, you are welcome to contact me off-line.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Herbststrasse 7 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: www.tvips.com Email: ingo.daberkow-at-tvips.com
-------- Original Message --------
Dear List -
We are preparing to embark on an in-depth project with outside funding that will require, among other things, 3-D reconstruction of series' of transmission electron micrographs of sections of biological material. We don't have a great deal of recent experience in this area and would welcome (off list if you are a commercial concern) suggestions for the "best" or most capable, or most flexible or useful 3-D reconstruction software including what computer platform is required and what configuration is considered adequate or better for this type of work. In addition, what digital data collection system currently available would be best for this type of work? We presently have a Philips CM12S STEM with a slow scan CCD camera with a 1k x 1k chip that has done good service for an extended period, but it seems that it might be a bit difficult to use for serial section recording and image orientation in preparation for 3-D reconstruction. If something new in a hardware-software package is available, we would be interested to hear how it works for others, or to hear directly from vendors off list.
Separately, it is possible that TEM tomography will also be in our future - It is our understanding that either at least IVEM or an energy filtered TEM are required to do useful tomography of thicker sections (0.25 - 0.5 micron thick). What are others in this area of interest using for microscopes, computers, and software? Has something of a "standard" come about, or are people still writing their own code and putting systems together from scratch?
Thanks in advance for any and all information and advice.
Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Never heard of them either but a quick internet search showed the following three sites:
1. Shimadzu Scientific Instruments, Inc. We provide solutions to analytical laboratory science by producing an extensive range of products, software and unrivaled customer service. In the United States, Shimadzu
http://www.ssi.shimadzu.com/
2. Shimadzu - Solutions for Science Shimadzu Scientific Instruments - Solutions for Science since 1875 http://www.shimadzu.com/
3. Shimadzu Solutions for Science We provide solutions to analytical laboratory science and medical science by producing an extensive range of products, software and customer service. In Europe, Shimadzu has a
http://www.sel.shimadzu.com/
====} As well as a specific South American Local site:
http://www.shimadzu.com.br
===================================
And since none of us seem familiar with Shimadzu I thought this site was interesting . .
4. Shimadzu SUCKS homepage Shimadzu specializes in lies, denials and cover-ups!
http://shimadzu.wxs.org/
(Now, take it for whatever its worth, but someone did go the effort and expense of setting up this site, eh?)
================================
On 21 Mar 2001, at 11:57, Leonardo Lagoeiro wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there, } } I would like to hearing from you opinions about Scanning microscopes. } We have a limited fund to buy a new scanning electron microscope and } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). I } would appreciate if someone out there could give me an opinion about } this equipment. Right now we can buy a unit without the EDS } analitical system which we are planning to buy later. Then I also } would like to know if we would buy only the image unit first we could } buy later the EDS system (from the same company) separately to } upgrade the same model (SS-550). The vendor said that it is possible, } but sometimes it is hard to trust in sellers especially when you live } in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
} ... } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). } ...
Any reputable SEM manufacturer should be able to provide you with a list of facilities with their SEM in use. If not, you should be wary about being the first.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
I use 1% glycine in PBS. Publish data : 0.15 M glycine or lysine.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Sara Miller {saram-at-duke.edu} 03/21 4:19 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ammonium chloride (50-100 mM) in PBS or glycine (sorry, don't remember concerntration--probably 1 or 2 % ??) in PBS have been reported. We use NH4Cl.
Sara Miller
On Wed, 21 Mar 2001, Caroline Miller wrote:
} Date: Wed, 21 Mar 2001 12:24:55 -0600 } From: Caroline Miller {camiller-at-creighton.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Aldehyde Removal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm looking for a method to remove aldehydes from tissue before } processing? Thanks, Caroline Miller } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
In my continuing quest to morph my lab from materials-oriented to bio-oriented I may finally get a chance to get a critical point drier or freeze drier. Could someone with more bio experience than I please help me out with some guidance? Or primary efforts will be intact bacteria and viruses although we may ultimately get into some minor amounts of tissue work. If I can only get either CPD or freeze drier which do I go with? is there any strong justification for both? what accessories are "must haves"? how much should I budget? and finally is there anything beyond the drier and accessories that I should be arranging?
I will not object if any suppliers reading choose to privately send a budgetary quote with the understanding that I am a US Govt. employee and am in no way authorized to negotiate purchases or commit resources.
Thank-you! Erica Valdes US Army SBCCOM Edgewood Chem Bio Center APG, MD 410-436-2608
To find them, you have to go to the Brasilian site, where there the SEM 550, the EPMA 1600, and an AFM are pictured on www.shimadzu.com.br/analitica/portugues/microscopia.htm
There's a page for the EPMA, seems to say 5 spectrometers www.shimadzu.com.br/analitica/portugues/microsonda.htm
Does anybody recognise it as a different brand?
Why only in Brazil?
Strange.
cheers
rtch
} Date: Thu, 22 Mar 2001 07:53:51 +0100 } From: Henrik Kaker {Henrik.Kaker-at-guest.arnes.si} } Reply-to: Henrik.Kaker-at-guest.arnes.si } Organization: Metal Ravne d.o.o., SEM-EDS Lab } To: Ritchie Sims {r.sims-at-auckland.ac.nz} , } "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com} } Subject: Re: SEM inquiry
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } } Hello, } } I was searched Shimadzu web site at http://www.shimadzu.com, but } there is no } } any information about scanning electron microscope. } } Henrik } } Ritchie Sims wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I didn't know that Shimadzu (who are a large and reputable scientific } } equipment manufacturer who afew years ago bought Kratos)) made an } } SEM, and a search of their website www.shimadzu.com revealed none. } } However, I see that their regional Brasil website lists not only the } } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } } may not have been so. } } } } I'd be very interested to hear more about either instrument. } } } } Anybody know anything? } } } } cheers } } } } rtch } } } } } } } } Hi there, } } } } } } I would like to hearing from you opinions about Scanning } } } microscopes. We have a limited fund to buy a new scanning electron } } } microscope and we have just received a proposal from a Japanese } } } company named SHIMADZU. They offered us a scanning electron } } } microscope model SS-550. I confess that I don't know much about this } } } company and neither about the quality of its instruments } } } (particularly SEM). I would appreciate if someone out there could } } } give me an opinion about this equipment. Right now we can buy a unit } } } without the EDS analitical system which we are planning to buy } } } later. Then I also would like to know if we would buy only the image } } } unit first we could buy later the EDS system (from the same company) } } } separately to upgrade the same model (SS-550). The vendor said that } } } it is possible, but sometimes it is hard to trust in sellers } } } especially when you live in South America. } } } } } } Thank all of you } } } } } } Best wishes, } } } } } } } } } Leonardo } } } -- } } } --- } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } } -- } Henrik Kaker, Ph.D. } Metal Ravne d.o.o. } SEM-EDS Lab } Koroska cesta 14, 2390 Ravne } Slovenia } Phone: +386 02 82 21 131 } Fax: +386 02 82 20 436 } http://www.kaker.com } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I've been asked to look into XRD equipment for our lab here, and was wondering if any of you know who the major players are in this area. We deal mostly with polymers and polymer fibers, and want to be able to do both small and wide angle work.
Thanks for the help. dz
David Ziegler U.S. Army, SBCCOM AMSSB-RSS-MS(N) Materials Science Team, SS&T Natick, MA 01760-5020 TEL: (508) 233-6484 FAX: (508) 233-5521 Email: David.Ziegler-at-Natick.Army.Mil
Dear Member of the Microscopy/Microanalysis Listserver,
following the very successful workshops at Lake Tahoe, Leukerbad and Port Ludlow, the next international workshop on electron energy loss spectroscopy and imaging will be held in Guadeloupe from May 5 - 9, 2002.
The abbreviated title will be
SALSA 2002
which stands for
'Strategies and Advances in Atomic Level Spectroscopy and Analysis'.
We think that this title very nicely summarises the central themes that will be discussed at the workshop and also suggests the stimulating environment in which the workshop will be held. You can find further announcements about the workshop on the new homepage
http://www.energyloss.com/
where you will also find a form to express your interest in SALSA 2002 (} This Meeting } Interested?).
May we kindly ask you to fill out the form with as much information as possible. This will help us to demonstrate the level of interest in the meeting in order to attract possible sponsors, including local authorities! If you fill in your e-mail address on the form, you will automatically be placed on our email distribution list and informed about future updates. Your e-mail address will only be used for information directly related to the workshop and will not be given to any other parties. If you prefer not to receive any future e-mails, please help us nonetheless by filling out the form and simply entering 'no' in the e-mail address field.
We hope to see you all at SALSA 2002!
The organisers:
Andrew Bleloch Christian Colliex Bernd Kraus Ondrej Krivanek Richard Leapman Jean-Louis Mansot Joachim Mayer David A. Muller Stephen J. Pennycook
Chris Jeffree wrote the following: ============================================================ Some questions arising: 1) The use of iridium for sputtering is news to me, and looks interesting. I would be interested to see a comparison of iridium film structure with platinum etc. Can a garden variety gold or gold/palladium unit sputter iridium, or are more exotic conditions required?
2) Does anyone have experience of the practical and safety issues arising from osmium coaters? Is the release of osmium tetroxide vapour to room atmosphere well controlled? How economical are they to run? Would they be practical in an environment where there could be several days or even weeks between coating runs? Presumably there are no safety issues with the coated specimens if the metal is inert. True? ============================================================== Chris raised two good points, one I can comment about with greater certainty than the other:
1] When sputtering precious metals with a typical ("garden variety") "SEM lab coater", with or without a turbo pump, there is a certain physics going on, and it is basically the same for all of the precious metals. Without going into a discussion of the nucleation and growth of a layer, the process is fundamentally the same, be it Au, Pt, alloys of these, or even Ir.
The mention of Ir in a previous posting was not in the context of a conventional SEM coater, and presumably there could be a different physics going on so the conclusion when coating is done in that equipment could be quite different (e.g. a much finer grain for example).
So while Pt in a conventional SEM coater does give a slightly smaller grain size, there are some trade offs (e.g. longer coating times) and in any case, it still does not satisfy one with a FESEM seeking to get the most out of their equipment.
2] We at SPI, in our demo lab have had one of the original OPC-40 Osmium Plasma Coaters operating for more than one year and then that was upgraded to the OPC-60 which is described on our website. Yes, the source of the osmium is osmium tetroxide, in 0.1g ampoules. For those not used to looking at osmium tetroxide in ampoules, a 1g amount of the material is described as being "1/3 of a teaspoon". So we are talking about pretty tiny amounts to begin with (e.g. 10% of 1/3 of a teaspoon). On the rotary vane mechanical pump pumping out the unit, in addition to the normal oil mist filter, on top of that is another filter, we call it an "osmium filter", probably a misnomer but it is designed to be a further filter of any mist. At the top is a white paper that serves as an indicator paper: the slightest amount of tetroxide getting out would turn the paper black.
So the first point is that this white paper is still the original paper that has been in continuous operation for more than two years. Translated this means that no tetroxide whatsoever is exiting the pumping system. Even when the unit is vented to atmosphere, there is a procedure where the chamber is first pumped out, and then the chamber can be opened. But one does not get a whiff of tetroxide vapor when the chamber is opened. Those seeing the demos at the shows would attest to that fact.
Of course, at some point there is some tetroxide being pumped out of the chamber, and if indeed some of the OsO4 did indeed get to the pump oil as tetroxide, it would be instantly reduced to the dioxide, something fairly innocuous, which ends up as a colloid in the pump fluid. So other than the apparent need to change the pump oil a bit more often, that seems to be the only consequence we have ever seen from the presence of the tetroxide.
With regard to the unit itself, it is completely safety interlocked, we have done literally hundreds of demonstrations at M&M, PITTCON, and other meetings and safety is the number 1 issue that comes up. But once anyone sees such a demonstration, there is agreement that safety no longer is an issue. It really is impossible for someone to get a whiff of the bad stuff when the chamber is opened.
Further information on this point would be the following: a) It is CE tested (passed the tests) for sale in Europe; these tests are among the most serious in the world in terms of safety. b) I have been in laboratories in Japan and they run the units out in the open although if one really was concerned about that, there is no reason why the unit could not be run in a fume hood.
Since there are well over 100 of the Nippon Laser coaters installed in Japan alone, I trust that there must be at least a few users of this equipment in Japan who might also feel qualified to comment on this approach to the coating of SEM samples. The unique technology embodied in the OPC-60 Plasma Coater is covered by US Patent #5855682; it makes interesting reading for one wanting to understand better the operation of this system.
3] The other issues raised had to do with cost and practicality. While the number of runs one could get depends of course on the thickness of the coatings being applied, from our own experience, using 0.1g ampoules, we typically get 20-40 runs per ampoule, so at a current cost of $8.00 per ampoule, the cost per run is indeed quite low, and in the ball park of cost one would associate with coating with gold. The capital cost is comparable to or less than most chromium coaters.
When the system is turned on, including the pump, the pump down is literally just a few minutes to coating. If the unit has sat unused for several months, we would expect that, as with any vacuum system, a bit longer for the pump down would be needed.
For the long term storage of osmium coated SEM samples, as with any delicate SEM sample, once prepared, we would recommend storing dry, not but not necessarily oxygen free.
Disclaimer: SPI Supplies is a distributor of the Osmium Plasma Coating equipment so we have a vested interest in seeing more coaters being sold. We would be happy to hear what others have to say about this system, pro or con.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Question up front: I¹d like to know how many labs go straight from 100% ethanol (no propylene oxide)to epoxy resin (both Spurr and Epon substitutes, or any other epoxies).
For years, we have deleted the propylene oxide step a.) to decrease the processing time and b.) to prevent having to deal with this toxic chemical. This works fine as long as you use dry ethanol, which we did by keeping drying beads (molecular sieves) in a small (200-500 ml) bottle of ethanol and oven-drying the beads when this absolute EtOH stock was depleted. We also made 3 changes of 100% ethanol (10-15 min each for small blocks, or longer for bigger pieces).
I have other folks in my realm that have routinely used propylene oxide; understandably, they don¹t want to change because it works. It would be nice to have one set of rules that always works. What I¹d like to know from you is how many of you have eliminated the PO, and are there instances where you must use it? Are there other procedures, such as using absolutely dry ethanol or extended times, that you follow if you use only ethanol?
Reply to me directly to save the net from hundreds of answers, and I will tabulate and send out a summary.
Thanks, Sara
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Hi All, I worked as a service engineer for Kratos after they became a subsidiary of Schimadzu. I installed a surface analysis system in Japan at the same time as a Schimadzu SEM with WDX detector was being installed in the room opposite. The system looked very well put together. Unfortunately since the installation engineer and customers spoke no English I have no specific information about the quality of the SEMs they make, but I can tell you they are a multibillion dollar corporation with headquarters in Kyoto, Japan and a significant presence in the US in facilities in Columbia, MD. Some of their products are only offered in Japan but they have an extensive range of analytical instruments that they do offer in the US.
Steve Buckingham Excellatron Solid State LLC 1640 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-muohio.edu] Sent: Thursday, March 22, 2001 9:51 AM To: microscopy-at-sparc5.microscopy.com
Leanardo etc.:
Never heard of them either but a quick internet search showed the following three sites:
1. Shimadzu Scientific Instruments, Inc. We provide solutions to analytical laboratory science by producing an extensive range of products, software and unrivaled customer service. In the United States, Shimadzu
http://www.ssi.shimadzu.com/
2. Shimadzu - Solutions for Science Shimadzu Scientific Instruments - Solutions for Science since 1875 http://www.shimadzu.com/
3. Shimadzu Solutions for Science We provide solutions to analytical laboratory science and medical science by producing an extensive range of products, software and customer service. In Europe, Shimadzu has a
http://www.sel.shimadzu.com/
====} As well as a specific South American Local site:
http://www.shimadzu.com.br
===================================
And since none of us seem familiar with Shimadzu I thought this site
was interesting . .
4. Shimadzu SUCKS homepage Shimadzu specializes in lies, denials and cover-ups!
http://shimadzu.wxs.org/
(Now, take it for whatever its worth, but someone did go the effort and expense of setting up this site, eh?)
================================
On 21 Mar 2001, at 11:57, Leonardo Lagoeiro wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there, } } I would like to hearing from you opinions about Scanning microscopes. } We have a limited fund to buy a new scanning electron microscope and } we have just received a proposal from a Japanese company named } SHIMADZU. They offered us a scanning electron microscope model } SS-550. I confess that I don't know much about this company and } neither about the quality of its instruments (particularly SEM). I } would appreciate if someone out there could give me an opinion about } this equipment. Right now we can buy a unit without the EDS } analitical system which we are planning to buy later. Then I also } would like to know if we would buy only the image unit first we could } buy later the EDS system (from the same company) separately to } upgrade the same model (SS-550). The vendor said that it is possible, } but sometimes it is hard to trust in sellers especially when you live } in South America. } } Thank all of you } } Best wishes, } } } Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } Organization: Dept of Geology, Univ of Auckland } To: Henrik.Kaker-at-guest.arnes.si, microscopy-at-sparc5.microscopy.com } Date: Fri, 23 Mar 2001 07:34:08 GMT+1200 } Subject: Shimadzu } Priority: normal
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } ---. } } } } Yes, that's what I said. } } To find them, you have to go to the Brasilian site, where there the } SEM 550, the EPMA 1600, and an AFM are pictured on } www.shimadzu.com.br/analitica/portugues/microscopia.htm } } There's a page for the EPMA, seems to say 5 spectrometers } www.shimadzu.com.br/analitica/portugues/microsonda.htm } } Does anybody recognise it as a different brand? } } Why only in Brazil? } } Strange. } } cheers } } rtch } } } } } } } } Date: Thu, 22 Mar 2001 07:53:51 +0100 } } From: Henrik Kaker {Henrik.Kaker-at-guest.arnes.si} } } Reply-to: Henrik.Kaker-at-guest.arnes.si } } Organization: Metal Ravne d.o.o., SEM-EDS Lab } } To: Ritchie Sims {r.sims-at-auckland.ac.nz} , } } "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com} } } Subject: Re: SEM inquiry } } } -------------------------------------------------------------------- } } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } ---. } } } } } } Hello, } } } } I was searched Shimadzu web site at http://www.shimadzu.com, but } } there is no } } } } any information about scanning electron microscope. } } } } Henrik } } } } Ritchie Sims wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } I didn't know that Shimadzu (who are a large and reputable scientific } } } equipment manufacturer who afew years ago bought Kratos)) made an } } } SEM, and a search of their website www.shimadzu.com revealed none. } } } However, I see that their regional Brasil website lists not only the } } } SEM 550, but also the EPMA 1600! I remember hearing, years ago, that } } } they marketed the ARL EPMA as a Shimadzu in (maybe) Asia, but that } } } may not have been so. } } } } } } I'd be very interested to hear more about either instrument. } } } } } } Anybody know anything? } } } } } } cheers } } } } } } rtch } } } } } } } } } } } Hi there, } } } } } } } } I would like to hearing from you opinions about Scanning } } } } microscopes. We have a limited fund to buy a new scanning electron } } } } microscope and we have just received a proposal from a Japanese } } } } company named SHIMADZU. They offered us a scanning electron } } } } microscope model SS-550. I confess that I don't know much about this } } } } company and neither about the quality of its instruments } } } } (particularly SEM). I would appreciate if someone out there could } } } } give me an opinion about this equipment. Right now we can buy a unit } } } } without the EDS analitical system which we are planning to buy } } } } later. Then I also would like to know if we would buy only the image } } } } unit first we could buy later the EDS system (from the same company) } } } } separately to upgrade the same model (SS-550). The vendor said that } } } } it is possible, but sometimes it is hard to trust in sellers } } } } especially when you live in South America. } } } } } } } } Thank all of you } } } } } } } } Best wishes, } } } } } } } } } } } } Leonardo } } } } -- } } } } --- } } } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } } Department of Geology Fax : 64 9 3737435 } } } The University of Auckland email : r.sims-at-auckland.ac.nz } } } Private Bag 92019 } } } Auckland } } } New Zealand } } } } -- } } Henrik Kaker, Ph.D. } } Metal Ravne d.o.o. } } SEM-EDS Lab } } Koroska cesta 14, 2390 Ravne } } Slovenia } } Phone: +386 02 82 21 131 } } Fax: +386 02 82 20 436 } } http://www.kaker.com } } } } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Ir as an ion beam sputtering target began in'92 after a scientist at Lucent Technology, formerly Bell Labs, was looking at silicon by STM. Before AFM, STM was the only type of SPM. In STM the tip contacts the specimen for tunneling. To increase specimen conductivity without introducing artifacts, Pt was ion beam deposited in an IBS, instrument developed by VCR GROUP now called IBS/e manufactured by SBT. Different metals were compared to Pt for minimum film resistivity and smoothness including Cr, Ti, W & Ir. Surprisingly Ir films prevented specimen surfaces from STM tip damage, were as conductive as Pt films, sputtered like Pt ie sputter rate & material distribution were the same and Ir does not oxidize.
After learning Bell Labs results, we hypothesized that if Ir films minimized STM tip damage, there may be a benefit to delicate specimens from electron beam interaction in FESEMs. Therefore thin films of Ir & Pt were deposited on 200 nm polystyrene spheres dispersed on mica. Several specimens with different metal thickness of Ir & Pt were examined in a Hitachi in-lens FESEM. It was observed that the PS spheres with Ir films showed less beam damage than those with Pt films. During separate TEM studies we learned that 5 to 100 angstrom thick Ir films had virtually identical structure as Pt films.
Further tests were done in Hitachi & JEOL FESEMs of thin Ir films on various substrates. No Ir structure, grains, could be seen below 200kX. Since '93, we have recommended Ir targets for the IBS since most FESEM work is carried out below 200kX. We still recommend Cr or other refractory metal targets for examination above 200kX.
Vince Carlino SBT, Inc.
Some questions arising: 1) The use of iridium for sputtering is news to me, and looks interesting. I would be interested to see a comparison of iridium film structure with platinum etc. Can a garden variety gold or gold/palladium unit sputter iridium, or are more exotic conditions required?
2) Does anyone have experience of the practical and safety issues arising from osmium coaters? Is the release of osmium tetroxide vapour to room atmosphere well controlled? How economical are they to run? Would they be practical in an environment where there could be several days or even weeks between coating runs? Presumably there are no safety issues with the coated specimens if the metal is inert. True?
vince thank you for this information. Iridium suppliers: Johnson Matthey Noble Metals Division supply it in various forms from facilities in Royston, Cambridge UK and West Chester, Pennsylvania USA. Sheet of 99.99% purity up to 10" wide by 0.1 to 5mm thick is available, and discs, wire etc. http://www.noble.matthey.com/product/detail.asp?id=34 An information sheet with contact addresses is available from the web site as a 47k .pdf file
Chris
----- Original Message ----- } From: {"VCRVINCE-at-aol.com"-at-sparc5.microscopy.com} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} Cc: {microscopy-at-sparc5.microscopy.com} ; "John F. Mansfield" {jfmjfm-at-umich.edu} Sent: Friday, March 23, 2001 1:00 AM
Folks, this is for a colleague of mine. If you have such a document please mail it to me at your earliest convenience Simon
I am seeking information appropriate to preparing an SOP to handle the eventuality that a mercury lamp explodes in one of our fluorescent microscopes. I am looking for sepcific information about potential personnel risks associated with the mercury vapor and what, if any, decontamination procedures are necessary/appropriate for potentially exposed equipment or personnel. I appreciate that the possibility of an explosion is remote but, I am required to address this eventuality.
Thank you again for sharing your findings with me.
------------------------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 3500 Terrace St University of Pittsburgh Pittsburgh PA 15261 Tel: 412-648-3051 Fax: 412-648-8330 URL: http://www.cbi.pitt.edu --------------------------------------------
I am in the market for a digital camera upgrade to my LaB6 2010. At the risk of receiving more information than I want to know about, venders are welcome to contact me off line to bring me up to date on the technology. Invitations to bid are expected to follow a review of the technology.
Thanks, Bruce Brinson Optical Analyst Rice University
Dear colleguages, I'm seeking a controller for a JSM840 whicsh is reliable and can control the SEM beam in X-Y direction at least with 4096 pixel resolution. And it would be nice if the stage and the controller would be optically coupled to the computer. I saw such a controller in some older letter, but I don't remember where? I thank any kind of help LAjos Pogany dr. Pogány Lajos Senior Research Fellow, Metals Research Department, Research Institute for Solid State Physics and Optics, Hungarian Academy of Sciences Office Address: H-1121 Budapest, Konkoly-Thege ut 29-33 Letters: H-1525 Budapest, P.O.B. 49, Hungary Phone: (00)-36-1-392-2222/17-25; Fax: (00)-36-1-392-2215 e-mail:pogany-at-szfki.hu
Announcement and Call for Papers TENTH CROATIAN-SLOVENIAN CRYSTALLOGRAPHIC MEETING Lovran, Croatia, June 21-24, 2001
The Meeting is open for the contribution dealing with all aspects of crystallography and its applications (Chemical Crystallography, Industrial Crystallography, Biological Crystallography, Physical Crystallography, Applied Crystallography and Mineralogy) and its techniques (XRD, TEM, SEM, STM)
please visit the web site
http://www.chem.pmf.hr/~hkz/lovran01/
Prof. A. Tonejc Department of Physics Faculty of Science Zagreb Croatia
In a message dated 03/23/2001 6:26:00 AM US Mountain Standard Time, swatkins+-at-pitt.edu writes:
{ { I am seeking information appropriate to preparing an SOP to handle the eventuality that a mercury lamp explodes in one of our fluorescent microscopes. I am looking for sepcific information about potential personnel risks associated with the mercury vapor and what, if any, decontamination procedures are necessary/appropriate for potentially exposed equipment or personnel. I appreciate that the possibility of an explosion is remote but, I am required to address this eventuality.
Thank you again for sharing your findings with me.
} }
Simon,
I know the manufacturers of the bulbs have this info. I have seen the precautionary statements on the flyers which accompany Osram bulbs, and Ushio probably has a similar document. If you don't hear from someone who already has this info, you could probably get it from Osram.
In a nutshell, Osram recommends that all personnel immediately leave the area for at least 30 minutes to avoid inhaling mercury vapor, and then any mercury residue, debris, envelope shards, etc. should be cleaned up with a mercury cleanup kit, and the debris properly disposed of as biohazardous material with mercury clearly identified as the hazard. Alternatively, if you have a hazmat team, they could be called in to do the cleanup, I suppose.
Hi! I am preparing a sample for TEM. The sample is GaN on sapphire substrate. I am supposed to make a sample in plane-view. Its needless to say how hard sapphire is. Grinding it down to 150microns was itself a tough job. Now I have put it on the dimpler, and using a diamond slurry to grind it. Its been 10 hrs and I have been successful in taking off only 70 microns. Does anyone have any bright ideas to speed up this process?
Praveena bhaskara Department of Chemical Engineering. University of Massachusettes, Lowell Lowell _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
One of my clients needs to do immunocytochemistry on mouse bone marrow, and needs to use frozen sections. I haven't got a clue where to begin (she will harvest the bone marrow). Any ideas?
Thanks as always, Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I'd just like to say "Thank You" to all of you who replied to my question on the EDS analysis of the contamiants. you answers proved to be most helpful.
Yep! The small angle cleavage angle works well for GaN on sapphire. I prepared about 8 samples in about 2 hours while teaching the technique to students and staff at the Univ. of Illinois. We only looked at three samples. Two were very very good. I will send you an image off-list. You can obtain information about South Bay Technology's Micro-Cleave kit at their web site, http://www.southbaytech.com
For grinding down the samples, use a 30 or 45 um diamond lapping film with a hand grinder. (I used 30um.) Works well.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Praveena Bhaskara [mailto:bubbyp-at-hotmail.com] } Sent: Friday, March 23, 2001 1:00 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM sample preparation } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hi! } I am preparing a sample for TEM. The sample is GaN on } sapphire substrate. I } am supposed to make a sample in plane-view. Its needless to } say how hard } sapphire is. Grinding it down to 150microns was itself a } tough job. Now I } have put it on the dimpler, and using a diamond slurry to } grind it. Its been } 10 hrs and I have been successful in taking off only 70 } microns. Does anyone } have any bright ideas to speed up this process? } } Praveena bhaskara } Department of Chemical Engineering. } University of Massachusettes, Lowell } Lowell } ______________________________________________________________ } ___________ } Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
} Dear Praveena: } } We have an application note on preparing GaN on Sapphire using our } Tripod Polisher. You can download it from our webiste at } www.southbaytech.com. Go to Applications Support then click on } Applications Notes. } } I hope this helps. } } David } } } ----------------------------------------------------------------------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi! } } I am preparing a sample for TEM. The sample is GaN on sapphire substrate. I } } am supposed to make a sample in plane-view. Its needless to say how hard } } sapphire is. Grinding it down to 150microns was itself a tough job. Now I } } have put it on the dimpler, and using a diamond slurry to grind it. Its been } } 10 hrs and I have been successful in taking off only 70 microns. Does anyone } } have any bright ideas to speed up this process? } } } } Praveena bhaskara } } Department of Chemical Engineering. } } University of Massachusettes, Lowell } } Lowell } } _________________________________________________________________________ } } -- } David Henriks TEL: +1-949-492-2600 } South Bay Technology, Inc. FAX: +1-949-492-1499 } 1120 Via Callejon } San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com } } *** www.southbaytech.com ***
Hg clean up kits are available from Lab Safety Supply, Box 1368, Janesville, WI 53547 (800 356 0783).
I have no commercial interest in this company. That's just where we got our kit. Fortunately, we haven't had to use it.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
---------- Forwarded message ----------
In a message dated 03/23/2001 6:26:00 AM US Mountain Standard Time, swatkins+-at-pitt.edu writes:
{ { I am seeking information appropriate to preparing an SOP to handle the eventuality that a mercury lamp explodes in one of our fluorescent microscopes. I am looking for sepcific information about potential personnel risks associated with the mercury vapor and what, if any, decontamination procedures are necessary/appropriate for potentially exposed equipment or personnel. I appreciate that the possibility of an explosion is remote but, I am required to address this eventuality.
Thank you again for sharing your findings with me.
} }
Simon,
I know the manufacturers of the bulbs have this info. I have seen the precautionary statements on the flyers which accompany Osram bulbs, and Ushio probably has a similar document. If you don't hear from someone who already has this info, you could probably get it from Osram.
In a nutshell, Osram recommends that all personnel immediately leave the area for at least 30 minutes to avoid inhaling mercury vapor, and then any mercury residue, debris, envelope shards, etc. should be cleaned up with a mercury cleanup kit, and the debris properly disposed of as biohazardous material with mercury clearly identified as the hazard. Alternatively, if you have a hazmat team, they could be called in to do the cleanup, I suppose.
I'm in another lab now, and in the hood here I found about 50 ml of DMF (or DMFA) - see below - and wondering what EMer's might use it for. Here's what the Merck Index says about it:
--------------------------- from the Merck Index:
DMF = N,N - Dimethylformamide (also abbreviated DMFA); C3H7NO Colorless to very slightly yellow liquid. Faint amine odor.
Human toxicity: Vapor harmful. Irritating to skin, eyes and mucous membranes. Liver injury has been produced in experimental animals by prolonged inhalation of 100 ppm.
Use: Solvent for liquids & gases. In the synthesis of organic compounds. Solvent for Orlon and similar polyacrylic fibers. Wherever a solvent with a slow rate of evaporation is required. Has been termed the universal organic solvent. ----------------------------
Any thoughts will be appreciated, thanks.
Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
To the person who was wondering weather or not P.O. was needed after dehydration and before embedding in Spurrs:
For the past 15 years I have been successfully embedding in Spurrs by using graded steps of acetone and going from my third change in 100% acetone to one third Spurrs and two thirds 100% acetone with out the use of P.O.. Of course then I follow that with a 1:1 mixture and then a 2:3 mixture and then a 100% Spurrs before embedding in 100% Spurrs at 65 degrees Celsius for 12 hours or more. Best of luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
I have been receiving conflicting reports concerning Amray service on the FE SEMs. The "official" word has been that Amray will continue to support the FESEMs "for some time" but I have had several customers call me & request service for their FESEMs as Amray will not offer a contract for the next year.
Has anyone had a similar experience or give me a striaght answer on this subject?
I was wondering if anyone has had any experience with lead aspartate "en Block" staining. I have tried "en block" staining before with disappointing results, but I have never tried it with lead asparate. The catalog describes it such that with Lead Aspartate you can then skip the staining step after sectioning.
Soft Imaging's ADDA is optically coupled between controlling computer and physical electrical interface to SEM.
http://www.soft-imaging.com
gary g.
At 08:18 AM 3/23/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Everyone, I want to make my own lacey carbon films for cryo-tem and was wondering if anyone had some good methods, experiences, insights, or references?
I found references for lacey formvar, pure carbon films, and a few others... But nothing explicitely for lacey carbon.
All comments greatly appreciated, Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Email: dfvillamar-at-acuabiotec.com Name: Daniel F. Villamar
Organization: AcuaBiotec LLC
Education: Graduate College
Location: Damascus, MD USA
Question: We are interested in SEM of samples of marine zooplankton, i.e., larval and postlarval shrimp. We would like to have a ventral view of the mouth parts and a view of the inside surface of the gut wall. The latter would require sectioning of the specimens along the longitudinal axis and scanning the inside surface starting at the mouth and working down through the foregut, midgut and hindgut of the specimens.
1. Does enyone provide such a service? 2. How should we fix and preserve the specimens for best result.
If I remember correctly, you make holey carbon films by first making your holey formvar (or other plastic), coating this film with a thin layer of carbon to make it continuous, and then dissolving the support plastic with a suitable solvent. This is off the top of my head. A few years ago, Mr. Pella suggested some literature to me on how to do it.
It's easier and more cost-effective to buy them from any of the EM supply houses!
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] } Sent: Friday, March 23, 2001 6:49 PM } To: microscopy-at-sparc5.microscopy.com } Subject: ? how to make lacey carbon films? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hello Everyone, } I want to make my own lacey carbon films for cryo-tem and was } wondering if } anyone had some good methods, experiences, insights, or references? } } I found references for lacey formvar, pure carbon films, and a few } others... But nothing explicitely for lacey carbon. } } All comments greatly appreciated, Thanks. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 } Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
The word I have received so far is that later model FESEMs will be covered under contracts. The distinction of "later model" is a perhaps nebulous term. The 1800 series instruments will probably not qualify. My 1910FESEM is based on 1800 electronics but uses the 305FE gun--and so far, is covered under contract. I am good for the rest of CY 2001. If Amray KLA stopped supporting FESEMs, something major would have to occur to keep the guns (at a minimum) available to established systems. The guns seem to last about 2.5-4 years. During that time, they are rock solid.
It seems to me that the more that a microscopist can do in regards to maintenance is better in the near term. This is despite a maintenance contract. If simple things go wrong and you can fix them, doing this will keep your system below the cost accountant's radar screen. The key for having the contract is if something really major goes wrong. An FE gun under contract is about $2K. Otherwise, it is at least $18K. How about a turbo pump? No cost under contract. Otherwise???
what about changing final apertures? I have not done a good job of doing this. But now I know more about how to do it correctly. And cleaning the holder is not a trivial process, nor one to be ignored.
gary g.
At 01:56 PM 3/23/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for TEM images for two reasons: 1. to become part of a digital image database for cell biology courses here at the University of British Columbia; 2. to be part of an image database used on CDs accompanying the new edition of a popular cell biology textbook.
I am looking for high quality images of a variety of cell types. These images can be either digitized or the original negatives. If they are digitized then they must be scanned-in to standards set by ourselves and the publisher. If the images are in the form of the original negatives we can arrange to have them sent here, scanned-in and returned.
In return contributors to the UBC collection can have access to other images in the collection and those who's images are used by the publisher will receive $50/US per image chosen. The publisher does not want to hold the copyright on the images used, rather they have proposed a generous licence agreement.
Interested parties can contact me (Brian Oates) at:
brian.oates-at-ubc.ca -- Dr Brian Oates Department of Botany University of British Columbia Vancouver, B. C. V6T 1Z4 604.822.4630 brian.oates-at-ubc.ca
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Snoring is the audible symptom of a {/FONT} {FONT color=3D"#0000A0"} {B} blocked airway during sleep. {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} The noise is caused by a vibration in the soft palate as the lungs pull ha= rd to take in a {/FONT} {FONT color=3D"#0000A0"} {B} weakened current of incoming air. {/B} {/FONT} {FONT color=3D"#0000A0"} This blockage may result from any number of circu= mstances, and these offer clues to get rid of the problem. {BR} {BR} Snoring is also more than likely to occur among people who {/FONT} {FONT color=3D"#0000A0"} {B} sleep on their backs {/B} {/FONT} {FONT color=3D"#0000A0"} ; the tongue falls back toward the throat and par= tly closes the airway. {BR} {BR} {/FONT} {FONT color=3D"#800040"} {B} The hidden dangers of snoring? {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {B} {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {UL} {LI} Snoring could increase the chances of a h= eart attack if not treated {BR} {LI} Hypertension and increased anxiety are associated with snoring {BR} {LI} Snoring causes sleep apnea a serious treatable medical condition {BR} {/LI} {/LI} {/LI} {/UL} {/FONT} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT color=3D"#800040"} {B} Snoring decreases sexual performance {/B} {/FONT= } {FONT color=3D"#800040"} {/FONT} {FONT color=3D"#0000A0"} by half, due to lack of oxygen to the brain which= decreases sexual responsiveness. {BR} {BR} Most people who snore have a {/FONT} {FONT color=3D"#800040"} {B} shortage of oxygen {/B} {/FONT} {FONT color=3D"#0000A0"} for a significant portion of that person's life.= In recent studies, daytime fatigue and cardiovascular disorder are relate= d to snoring. In a few cases, heavy snoring is a sign of potentially life = -threatening problem. Sleep apnea in which breathing stops for a second or= even minutes at a time and finally resumes with snorting and tossing arou= nd. This pattern may be repeated hundreds of times all night long. The res= ult is chronic oxygen shortage, which leads to abnormal heart rhythms, hig= h blood pressure, and heart strain. {BR} {BR} {/FONT} {FONT color=3D"#800040"} {B} How the Snore Eliminator works! {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} Snore Eliminator offers a unique break through wi= th its all {/FONT} {FONT color=3D"#800040"} natural enzymes {/FONT} {FONT color=3D"#0000A0"} which metabolize the secretions, allowing the bo= dy to absorb them into the back of the throat. The herbs reduce tissue swe= lling. The result is to open the airway and smooth the airflow. This in tu= rn eliminates the snoring for hours at a time. It also {/FONT} {FONT color=3D"#800040"} {B} reduces the noise {/B} {/FONT} {FONT color=3D"#800040"} made by breathing through the mouth while sleepi= ng {/FONT} {FONT color=3D"#0000A0"} . By simply spraying the back of the throat, tong= ue and uvula with our special formula, manufactured using our patented bre= ak through process, the soft tissue is coated. This allows the {/FONT} {FONT color=3D"#0000A0"} {B} {/B} {/FONT} {FONT color=3D"#800040"} {B} throat a chance to relax {/B} {/FONT} {FONT color=3D"#0000A0"} thus a reduced amount of snoring. In most cases = snoring is eliminated. Snore Eliminator coats the throat area with a uniqu= e patented process of all natural lubricants. {BR} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} TESTIMONIALS from several of our satisfied cus= tomers: {BR} {BR} {/B} {/FONT} {FONT face=3D"Times New Roman"} {FONT color=3D"#FF0000"} {B} Lorenzo writes... {BR} {BR} I am having a good nights sleep, finally. I used to wake myself up during= the night which was a real bummer as I would not always go back to sleep.= Sleeping through the night is great. Thanks a lot Snore Eliminator. {BR= } PS: My girlfriend loves it as much or more than I do. {BR} {BR} Alex writes... {BR} {BR} My husband uses it for another reason. He has a problem with blocked nasa= l passages. While he's asleep the passages close, he stops breathing and = he wakes up gasping. When he takes Snore Eliminator his nose stays cleare= r and he's able to sleep more restfully. {BR} Thank you for a product that has really helped us! {BR} {BR} Glen writes... {BR} {BR} In February 1995, I was diagnosed with severe sleep apnea. A CPAP was the= suggested treatment for my affliction, but it did not help my condition. = I heard your advertisement and ordered a bottle of Snore Eliminator and m= y sleep improved immediately. I continue to use it and my sleep has been = better than it had been in years. Thanks. {BR} {BR} Steve writes... {BR} {BR} Let me take this opportunity to thank you for your wonderful product and t= ell you how much my wife, Tracy, and I appreciate the quiet nights we have= enjoyed since discovering it. It's very rare when a product actually del= ivers what it promises but Snore Eliminator truly does. We have tried most= of the other advertised remedies without any satisfaction and I am very p= leased to be able to write and say that this product is as close as you ca= n get to an absolute "Miracle". {BR} {BR} {/B} {/FONT} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#800040"} If you have a snoring problem what can you do? {/F= ONT} {FONT color=3D"#0000A0"} Who would you turn to? Look no further, the ans= wer to your snoring problems are over. Let our patented {/FONT} {FONT color=3D"#0000A0"} {B} Snoring Eliminator {/B} {/FONT} {FONT color=3D"#0000A0"} help you get through those sleepless nights. We = guarantee it. In fact, we are so confident in Snore Eliminator that if you= are not 100% satisfied with it you can return the unused portion for a fu= ll refund, no questions asked* {BR} {BR} It is estimated that 90 million Americans, over the age of 18,suffer from = habitual snoring. {BR} {BR} {/FONT} {FONT color=3D"#800000"} {B} How to Use Snore Eliminator: {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} Open your mouth {BR} Hold the 4oz bottle about four inches from your open mouth {BR} Spray one or two application onto the back of the throat. {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} {BR} Snore Eliminator is safe to use nightly because of its' natural ingredient= s. {BR} Please note as with any health-related product, it is recommended that you= consult with your physician prior to use if you have any medical conditio= ns. {BR} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} A Money Back Guarantee! * {BR} {/B} {/FONT} {FONT face=3D"Times New Roman"} {BR} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#0000A0"} {B} We are so confident in Snore Eliminator that w= e offer a 15-day money back guarantee. Merely return the unused portion o= f Snore Eliminator within 15 days and we will refund your purchase price w= ith no questions asked! {BR} {BR} Our normal price for Snore Eliminator is $29.95, but for a limited time on= ly, you can purchase it for only {/B} {/FONT} {FONT color=3D"#800040"} {B} $19.49 plus $3.95 {/B} {/FONT} {FONT color=3D"#0000A0"} {B} for shipping handling. We will also continue t= o offer you Snore Eliminator at this low price as long as you purchase fro= m us in the future. To take advantage of this savings you must order with= in the next 10 days. {BR} {BR} Here's How to order {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} You can order by mailing us a Check, Cash or a Money Order. You can also = order by faxing us a copy of your Check. Faxing your order to us speeds y= our order up by 3-5 days. {BR} {BR} To order by Check, Cash or Money Order merely send $19.49 plus $3.95 for a= total of $23.44 per bottle. 1 bottle of Snore Eliminator will last you 4-= 6 weeks. Order TWO bottles for $19.49 and we do not charge you any shippin= g and handling. This means you will receive an 8 to 12 week supply of Sno= re Eliminator for only $38.98, including shipping and handling. This is a = 17% savings. {BR} {BR} Mail your check, cash or Money Order along with your name address, phone n= umber and email address filled out on the following form: {BR} {BR} Your Name:___________________________________________(please print) {BR} {BR} Street: ______________________________________________ {BR} {BR} City:______________________State/Province:____________________ {BR} {BR} Zip/Postal Code:_________________Country:_________________________ {BR} {BR} Phone number:______________________________________________ {BR} {BR} FaxNumber:_________________________________________________ {BR} {BR} Email address:_______________________________________ {BR} (if there is a problem with your order this is where we will notify you) {B= R} {BR} To: {BR} {BR} John Taylor {BR} CEO {BR} Internet Information Services, Snore Eliminator Division {BR} PO Box 21442 {BR} Billings, MT 59104 {BR} {BR} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} {BR} To Fax us your Order (Fill out and fax us every page which follows): {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT color=3D"#0000A0"} {B} MAKE YOUR CHECK PAYABLE TO {/B} {/FONT} {FONT color=3D"#0000A0"} : Internet Information Services, Snore Eliminator= Division, tape your check to the following form and Fax to {/FONT} {FONT color=3D"#0000A0"} {B} 1-775-599-3092 {/B} {/FONT} {FONT color=3D"#0000A0"} . If this line is busy, please try faxing to {BR= } {/FONT} {FONT color=3D"#0000A0"} {B} 1-801-383-9082. {/B} {/FONT} {FONT color=3D"#0000A0"} Faxing your check to us speeds your delivery up = by 3-5 days. {BR} {BR} Please ship me 1 bottle of Snore Eliminator for $19.49 plus $3.95 shipping= and handling for a total of $23.44 per bottle. Order TWO bottles for $19= .49 and we do not charge you any shipping and handling. This means you wi= ll receive an 8 to 12 week supply of Snore Eliminator for only $38.98, inc= luding shipping and handling. This is a 17% savings. {/FONT} {FONT face=3D"Times New Roman"} {FONT color=3D"#0000A0"} {BR} {/FONT} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#0000A0"} {BR} Your Name:___________________________________________(please print) {BR} {BR} Street: ______________________________________________ {BR} {BR} City:______________________State/Province:____________________ {BR} {BR} Zip Code:_________________Country:_________________________ {BR} {BR} Phone number:______________________________________________ {BR} {BR} Fax Number:_________________________________________________ {BR} {BR} Email address:_______________________________________ {BR} (if there is a problem with your order this is where we will notify you) {B= R} {BR} ************************************************************** {BR} {BR} Internet Information Services Check Authorization and Order Form {BR} {BR} I wish to authorize the purchase of the Snore Eliminator from Internet Inf= ormation Services using this order and check authorization form. I hereby= authorize Internet Information Services to duplicate the attached check i= n bank draft form for the amount shown on this attached check. I will ret= ain my original copy for my record of this transaction. {BR} {BR} This Authorization is valid for this transaction only. No other bank draf= ts may be created without my direct written authorization. {BR} {BR} Dated:_________________________________ {BR} {BR} Signed:________________________________________ {BR} {BR} {BR} .Tape your check here and Fax it to us... {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} {BR} Fax this form to: {/FONT} {FONT color=3D"#0000A0"} {B} 1-775-599-3092 {/B} {/FONT} {FONT color=3D"#0000A0"} . If this line is busy, please try faxing to {/FO= NT} {FONT color=3D"#0000A0"} {B} 1-801-383-9082. {BR} {BR} {/B} {/FONT} {FONT color=3D"#0000A0"} Thank you for your business, {BR} {BR} John Taylor {BR} CEO {BR} Internet Info Service, Snore Eliminator Division {BR} PO Box 21442 {BR} Billings, MT 59104 {BR} {BR} {BR} {BR} {BR} {BR} {/FONT} {FONT face=3D"Times New Roman"} {FONT color=3D"#000080"} {BR} {BR} {BR} {BR} {BR} {/FONT} {FONT face=3D"Courier New"} {FONT size=3D3} {FONT color=3D"#000000"} +++++++++++++++++++++++++++++++++++++++++++++++++= +++++++ {BR} This ad is produced and sent out by: {BR} Universal Advertising Systems {BR} To be removed from our mailing list please email us at {BR} vanessagreen-at-freeze.com with remove in the subject line or {BR} call us toll free at 1-888-605-2485 and give us your email address or writ= e us at: {BR} Central DB Removal, PO Box 1200, Oranjestad, Aruba {BR} ++++++++++++++++++++++++++++++++++++++++++++++++++++++++ {/FONT} {FONT face=3D"Times New Roman"} {FONT size=3D3} {BR} {FONT face=3D"MS Sans Serif"} {FONT color=3D"#000000"} {BR} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/FO= NT} {/FONT} {/FONT} {/FONT} {/FONT} {/FONT} {/BODY} {/HTML}
Michael Reiner University of Cologne, Germany Dept. of Anatomy I
} Hi, } } I'm passing on a question from a colleague. Does anyone know of a technique } for etching LR White from sections on a slide? I gave him all the info I had } on epoxy removal using ethoxide and so on, but I've been unable to locate } any references specific to LR White. } } Thanks. } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } }
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I am negotiating with my college to purchase a block of time on their new variable pressure SEM (Hitachi S-3000N with EDX). We have agreed to abide by the 'average' cost per hour charged by universities and colleges. I seem to remember that there was a discussion along these lines some time ago but I cannot find it in the archives. If you pay per-hour fees for SEM usage I would be much obliged if you would let me know what they are.
Thanks
Michael
Michael McInerney Rose-Hulman Institute, Terre Haute, IN.
Amray customers, as well as the rest of us, are scrambling to figure out a lot of things. I've had loyal Amray customers running to me because they've had their service dropped, they've had implied threats that their service would be dropped in the near term and because they are just sick and nervous of the way the whole thing was handled by the manufacturer. I am quite sure that there are many who are just plain confused over whether they have been dropped or not.
On the other hand, a new customer of mine had to field a question from an Amray rep over whether they were renewing a contract on a 3300 FESEM. When told they were not, the rep apparently went over the line about the 'impossibility' of getting other service and the 'proprietary' nature of their service.
I think that, having punched a hole in their own boat, Amray is now surprised that they are taking on water.
There are a lot of long-time, dedicated people at Amray who feel betrayed by their own company now. I know, having been through a very similar business turn with ETEC when they were bought out by Perkin-Elmer and their SEM business was left to suffer an ignoble death. It's hard to believe that anyone could possibly have made such decisions without having a long term desire to shutdown the business.
The only question that seems to remain is - what business is being shut down? Amray has always been a major player in the semi-conductor field. These days, the FESEM is the most attractive portion of that business. My guess would be that they have chosen to still pursue the semi manufacturers with FESEMs but close down their general purpose SEM business. In the mean time, they want to still provide the lucrative service contracts on the general purpose FESEMs out there. Cash is king, and the cash in their operation is in the selling and service of FESEMs, primarily to those customers who are willing and able to pay a premium for these instruments.
Overall, this move could only be construed as an attempt to increase the profit margin of their service operation. The general purpose SEMs are spread out geographically around the country, where the semi business is far less so. The older instruments represent an additional training cost for new service people. By limiting the number of instruments being serviced, the geographical spread of those instruments and the learning curve of their service staff, they can drastically cut back their service staff, weather attrition and defections of service staff by having new hires online quicker and in the end, probably have a staff that is paid less to do more. The same could also be said of their sales force.
Don't sweat, Gary - cathode emitters are available for the Amray FESEMs in the aftermarket for around $2,500 and turbopump exchanges cost about the same from the manufacturer. I can't really address the pricing strategies of other service providers, but in my case, whatever type of contract you choose, or even on a billable basis, you would save at least a few thousand a year over the Amray service costs. I'll cover the cathode by tacking on the $2,500 per year to the contract and simply change it once a year, and still beat Amray's cost by $4,000 - $6,000 per year. But I suggest that the customer cover it themselves - buy a new emitter when they first take out a contract and keep it on hand. In this way, the customer can themselves better control the cost through proper care and operation of the instrument, and deciding for themselves when a replacement is needed. This also gives you that warm fuzzy feeling knowing that you have one standing by, ready to use if needed. Third party service providers have great flexibility in writing contracts - in the case above, I would normally cover the cost of installation of the emitter even though you may have chosen to be responsible for the emitter.
Your mention of those using the instrument having some ability to take care of many problems they run across is right on the mark. I always prefer customers who have a real interest and knowledge of the instrumentation they use. Not because it may make my life easier (although it does because I know I'm talking to someone knowledgeable and am more willing to take what they say at face value), but because it's fun to share with, and learn from, those with similar interests.
I'm all for having a customer watch over my shoulder and understand what I'm doing, because it protects their better interests as well as mine. I'm not perfect, and sometimes (perhaps often) I'll get off on the wrong direction on a problem. The better the description of the problem I get, the better I'll do. It also helps sometimes to have a back-seat driver to let you know when you've taken a wrong turn, as damaging to the ego as that can be. Of course, for the customer, having a better knowledge of the instrument and its service means some peace of mind in finding someone to service it.
Kind of like knowing something about the various systems and construction methods in a house before hiring a remodeling company.
On Friday, March 23, 2001 8:32 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } } The word I have received so far is that later model FESEMs will be } covered under contracts. The distinction of "later model" is } a perhaps nebulous term. The 1800 series instruments } will probably not qualify. My 1910FESEM is based on 1800 } electronics but uses the 305FE gun--and so far, is covered } under contract. I am good for the rest of CY 2001. If Amray } KLA stopped supporting FESEMs, something major would have } to occur to keep the guns (at a minimum) available to established } systems. The guns seem to last about 2.5-4 years. During } that time, they are rock solid. } } It seems to me that the more that a microscopist can do } in regards to maintenance is better in the near term. This } is despite a maintenance contract. If simple things go } wrong and you can fix them, doing this will keep your } system below the cost accountant's radar screen. The key } for having the contract is if something really major goes } wrong. An FE gun under contract is about $2K. Otherwise, } it is at least $18K. How about a turbo pump? No cost } under contract. Otherwise??? } } what about changing final apertures? I have not done a good } job of doing this. But now I know more about how to do it } correctly. And cleaning the holder is not a trivial process, } nor one to be ignored. } } gary g. } } } At 01:56 PM 3/23/2001, you wrote: } } } } } Hi Listers, } } } } I have been receiving conflicting reports concerning Amray service on the FE } } SEMs. } } The "official" word has been that Amray will continue to support the FESEMs } } "for some time" but I have had several customers call me & request service } } for their FESEMs as Amray will not offer a contract for the next year. } } } } Has anyone had a similar experience or give me a striaght answer on this } } subject? } } } } } } Thank You, } } } } Earl Weltmer } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} [snip] } } The only question that seems to remain is - what business is being shut } down? Amray has always been a major player in the semi-conductor field. } These days, the FESEM is the most attractive portion of that business. My } guess would be that they have chosen to still pursue the semi manufacturers } with FESEMs but close down their general purpose SEM business. In the mean } time, they want to still provide the lucrative service contracts on the } general purpose FESEMs out there. Cash is king, and the cash in their } operation is in the selling and service of FESEMs, primarily to those } customers who are willing and able to pay a premium for these instruments.
Amray, as a company, no longer exists. Amray is a division of KLA-Tencor. For some reason, Amray apparently could not compete in the general purpose (lab) SEM marketplace. Or, other forces came into play which caused the change.
I do know that Amray had developed very good FESEM systems--optics, control software, stages, etc. KLA-Tencor is in the semiconductor defect analysis business. They lacked the type of SEM technology that Amray had. Consequently, KLA-Tencor purchased Amray, ostensibly I believe, to acquire Amray's SEM technology. This is similar to the relationship between FEI and Philips, wherein Philips acquired FEI for its ion beam expertise. To wit, FEI now produces a fine dual beam focused ion beam system which combines ion beam technology and SEM imaging (e.g., FEI 830).
KLA-Tencor produces a top line semiconductor defect analysis tool set (8100xl, 8300, 8500, etc.) which combines KLA's best attributes along with the FESEM qualities of Amray. The defect analysis tools cost about $2M each, compared to a typical lab FESEM at say $400K. The earlier Amray systems were priced lower than this. It seems to me that KLA, with Amray in tow, had made a decision to focus on the high cost (high profit) business area of semiconductor defect analysis, at the exclusion of lab SEMs. That is why we are seeing what is happening now with Amray service contracts.
In some respects, this was a myopic decision. Currently, the semiconductor industry is in a decline (albeit not monumental). There surely was and is a market for lab SEMs. KLA obviously made a business decision to focus on semiconductor defect analysis tools. From what I am seeing, the older Amray systems are no longer being supported by Amray. Since the thermionic systems are rather simple in design and easier to maintain than FESEMs, this may not be too bad. But, if one had a bad PC-10, that is of course a different matter.
So far as I can tell, Amray will support the FESEMs. Why they would not support a 3300 eludes me. Perhaps that is the bow wave of non-support for all Amray systems. I don't know. But I do know that a new Philips, Jeol, or Hitachi system at say $400K+ is about triple what I have invested in my current Amray FESEM. As long as my Amray system does what I need and can be maintained to do it, it makes no economic sense for me to dump it for a horribly expensive replacement. I suppose that what may happen is that the few lab SEM makers will offer about the same compliment of systems at about the same price mix. And the prices will be painful.
I have seen this same type of consolidation in the UK and other countries. For us Amray owners, it is a real problem....conundrum?
----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Gary Gaugler'" {gary-at-gaugler.com} ; "Earl Weltmer" {earlw-at-pacbell.net} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, March 24, 2001 3:31 PM
Didn't mean to leave you with any misconceptions. I'm aware of the purchase by KLA and as far as I am aware, any decisions made were theirs. To date, as far as I know, sales have still been under Amray brand name, which is why I referred to them that way. I believe that there were some non-general purpose SEM technologies that KLA was primarily interested in and will roll those into its own production. Perhaps Amray was poised to take a firmer stance in those areas? As far as Amray, it has always been more of a family run business and perhaps the offer was right and the time was right. I don't think there were any particular problems in the operation or bottom line of Amray.
I also didn't mean to leave anyone with the impression that they are not supporting the 3300. On the contrary, what I wrote was that they were upset that anyone would consider a third-party service provider for one. They were quite upset that they had lost one.
On Saturday, March 24, 2001 9:34 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } At 03:31 PM 3/24/2001, you wrote: } } } [snip] } } } } The only question that seems to remain is - what business is being shut } } down? Amray has always been a major player in the semi-conductor field. } } These days, the FESEM is the most attractive portion of that business. My } } guess would be that they have chosen to still pursue the semi manufacturers } } with FESEMs but close down their general purpose SEM business. In the mean } } time, they want to still provide the lucrative service contracts on the } } general purpose FESEMs out there. Cash is king, and the cash in their } } operation is in the selling and service of FESEMs, primarily to those } } customers who are willing and able to pay a premium for these instruments. } } Amray, as a company, no longer exists. Amray is a division of KLA-Tencor. } For some reason, Amray apparently could not compete in the general } purpose (lab) SEM marketplace. Or, other forces came into play which } caused the change. } } I do know that Amray had developed very good FESEM systems--optics, } control software, stages, etc. KLA-Tencor is in the semiconductor defect } analysis business. They lacked the type of SEM technology that Amray } had. Consequently, KLA-Tencor purchased Amray, ostensibly I believe, } to acquire Amray's SEM technology. This is similar to the relationship } between FEI and Philips, wherein Philips acquired FEI for its ion beam } expertise. To wit, FEI now produces a fine dual beam focused ion beam } system which combines ion beam technology and SEM imaging (e.g., FEI 830). } } KLA-Tencor produces a top line semiconductor defect analysis tool set } (8100xl, 8300, 8500, etc.) which combines KLA's best attributes along } with the FESEM qualities of Amray. The defect analysis tools cost about } $2M each, compared to a typical lab FESEM at say $400K. The earlier } Amray systems were priced lower than this. It seems to me that KLA, } with Amray in tow, had made a decision to focus on the high cost (high } profit) business area of semiconductor defect analysis, at the exclusion } of lab SEMs. That is why we are seeing what is happening now with } Amray service contracts. } } In some respects, this was a myopic decision. Currently, the semiconductor } industry is in a decline (albeit not monumental). There surely was and } is a market for lab SEMs. KLA obviously made a business decision } to focus on semiconductor defect analysis tools. From what I am seeing, } the older Amray systems are no longer being supported by Amray. } Since the thermionic systems are rather simple in design and easier } to maintain than FESEMs, this may not be too bad. But, if one had } a bad PC-10, that is of course a different matter. } } So far as I can tell, Amray will support the FESEMs. Why they } would not support a 3300 eludes me. Perhaps that is the bow wave } of non-support for all Amray systems. I don't know. But I do know } that a new Philips, Jeol, or Hitachi system at say $400K+ is about } triple what I have invested in my current Amray FESEM. As long } as my Amray system does what I need and can be maintained to do it, } it makes no economic sense for me to dump it for a horribly } expensive replacement. I suppose that what may happen is that } the few lab SEM makers will offer about the same compliment of systems at } about the same price mix. And the prices will be painful. } } I have seen this same type of consolidation in the UK and other } countries. For us Amray owners, it is a real problem....conundrum? } } gary g. } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Question: I recently purchased a microscope used in forensics with a state agency. I would like to study microscopy so I could show this fascinating subject to my grandchildren. As a youngin' I enjoyed biological and petrology studies with the microscopes. I'm looking for a course that would allow me to use the different features of the microscope in light and dark field applications. I'm retired and would appreciate any help in this endeavor that you could offer. I am suffering from a physical disability and erudition is how I go about studies these days. THANX again.
For a fairly comprehensive list try http://www.xraysite.com/index.html?producers.html
I have experience with diffractometers made by Rigaku, Scintag and Siemens - all have good systems.
I have also dealt with Jim Stauver at JSTechnical. He sells and services Siemens systems and can probably get you a good deal on a used system. His web site is http://www.jstechnicalservices.com/
If you need any more info or help, contact me off-list.
Sincerely, James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction (214) 403-6342 visit my web site at http://www.ktgeo.com/
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } Hello listers } } I've been asked to look into XRD equipment for our lab here, and was } wondering if any of you know who the major players are in this area. We } deal mostly with polymers and polymer fibers, and want to be able to do both } small and wide angle work. } } Thanks for the help. } dz } } David Ziegler } U.S. Army, SBCCOM } AMSSB-RSS-MS(N) } Materials Science Team, SS&T } Natick, MA 01760-5020 } TEL: (508) 233-6484 } FAX: (508) 233-5521 } Email: David.Ziegler-at-Natick.Army.Mil
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Larry -
You're not lilely to find a "course" that will meet your needs. Try starting with this book; this listing is taken from the Project MICRO bibliography, which you will find at http://www.msa.microscopy.com/ProjectMICRO/PMBooks.html
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy; it's unequalled as a basic reference for beginners. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with good illustrations) of commonly encountered organisms. Adult. RECOMMENDED
If you want a more sophisticated, interactive (and more expensive) approach, try this CD-ROM (from the same bibliography):
Pagliaro, l., Murray, C., Curran, G., Orkand, A., and Astion, M. 1997 Microscopy-Tutor. ISBN 0-7817-1217-3 $195.00 plus shipping; distributed by Lippincott-Raven Publishers, 12105 Insurance Way, Hagerstown, MD 21740, 800-638-3030. For Macintosh and Windows 3.1 or 95/NT; easy installation. Developed by the Department of Laboratory Medecine and the Center for Bioengineering at the University of Washington, Seattle, this is a college-level introduction to the use of a research-quality compound microscope. Its extensive use of QuickTime animation to illustrate alignment steps and optical principles make it much more than a "book on a CD"; moving ripples on a pond really do make it easier to understand wave theory! Kohler illumination is emphasized and explained. Most, but not all, terminology is defined; a glossary would help beginners. Proper care of lenses is covered well, but there's no instruction on how to use immersion oil properly. There is a brief concluding self-test that is more of a review of important information than a quiz. Although it's a good CD, it's definitely not appropriate for precollege microscopists. Adult.
And there are dozens of things listed for the grandkids...
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Having advised one of our customers to try the above resin (glutaraldehyde/carbohydrazide mixture) to embed her particularly sensitive material for LM (it was dissolving using other mainstream embedding media), she has now successfully got to the sectioning stage. Next problem is getting the sections to stick to the microscope slide. They curl and crack and disappear Lemming like into the abyss.
Having cut resin sections for many years I have now exhausted all the tricks that I have used to keep them on the slide from poly-L-lysine to silane to drying in an oven overnight (instead of using a hotplate or slide dryer). That last advice has surprisingly saved many desperate microscopists but not this time. Maybe one of the adhesive sticks which lays down a sticky membrane will work. We will even try double sided tape (or maybe not), but does anyone out there have experience with sections cut from this material? Help,
Regards
Terry Cooper TAAB Laboratories Equipment Ltd England
Thanks to all who offered help on the subject of the high resolution image if the MSA logo. Most thanks go to Bev Maleef who provide the goods.
I did have someone offer to take a micrograph of it with an SEM! I didn¹t know we actually had a copy ofthe MSA logo that small, has any one been playing the STM tricks with the individual atoms a la IBM?
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
Greetings, I have found that polyethlyene-imine (PEI) is much stickier than poly-lysine. I have no idea about the resin you mentioned, but PEI is sticky stuff. It comes as a liquid. I make a 0.1% soloution in ddwater, which I freeze in aliquots. Then I keep a working one in the fridge. I coat coverslips in the stuff by floating them on a drop of the PEI solution for about 10 sec, and then blotting off the excess and letting them air dry. In that conditions, the coated 'slips are good for at least months.
Hope this helps, Tobias Baskin
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Hi all, Can anyone in the Atlanta area help with this request? Please send your replies to Ulrika - E-mail: Ulrika.Egertsdotter-at-ipst.edu. thanks for your help, Beth Here's the request: } I am an } assistant scientist currently working at the Institute of Paper science and } technology in Atlanta and I am trying to find a cryomicrotome that I could } use for my project. Within my project, we are broadly speaking trying to } reveal information about the biological mechanisms of wood formation. We } are using different types of arrays, and for the sample preparations we are } trying to get as thin sections as possible. For this purpose I am now } looking for a cryomicrotome that I could access to make thin sections of } wood samples that would then be processed for RNA to probe the arrays, and } also hopefully in situ hybridisation. I am planning to go to Sweden to } learn the techniques from the group that has already been doing this type } of work in poplars. I will however need to find an accesible cryomicrotome } to work with that are somewhat closer to Atlanta. Do you think it would be } possible at all for me to make use of the cryomicrotome in your department, } or would you perhaps know of any other available equipment? } } Thank you. } } Sincerely, } Ulrika Egertsdotter } E-M Ulrika Egertsdotter, Ph.D. } Institute of Paper Science and Technology } 500 10th Street, N.W. } Atlanta, GA 30318 } } Direct dial: +1 404 894 7842/4807 } Telephone: +1 404 894 5700 } Fax. +1 404 894 4778 } E-mail: Ulrika.Egertsdotter-at-ipst.edu
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I am very new to electron microscopy, so I ask your indulgence in some basic questions.
I have an old JEOL SEM that has been sitting at atmospheric pressure for some time now and will need a thorough baking. It has a plain old tungsten wire filament, so the vacuum requirements aren't all that stringent. The SEM has rubber o-rings (I don't know how to tell if they are Viton). I doubt that I want to go above 100 oC.
1) What characteristics are important in choosing a heat tape suitable for baking the instrument?
2) Where would I find a suitable heat tape (my web searches on heat tapes got me lots of information about ice buildup on my roof, but very little about getting water out of an SEM)?
3) How do I determine how much to buy - how much column on each side of the tape can I assume to be heated properly by the tape - what is an appropriate center-to-center distance between adjacent wraps of tape?
4) Is there a more practical method than heat tapes for baking the instrument?
5) What else may I be missing here?
Thank you,
Bruce Girrell in snowy northern Michigan with a few more basic questions waiting in the wings
The Sudan dyes are those most commonly used for suberin, and perhaps cutin. The lignin component of both can be stained with phloroglucinol (or observed as autofluroescence with blue excitation in unstained material). If the autofluroescence is quenched by the Sudan, this supports the interpretation that the substance is suberin or cutin rather than lignin.
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R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility, Associate Member Donald Danforth Plant Science Center/Nidus Center 893 North Warson St. Louis, MO 63141
Would anyone know of a general protocol I could use to stain a nylon 6,6 sample for TEM analysis? I'm studying cross-sections of nylon fibers. I've read that tin chloride is a good stain for nylons, I've also seen reference to work using iodine.
I would not recommend baking out that system if it has not been designed for it. The heat tapes will give you hot spots and other places may not take the heat well. If you are not going above 100C, it is not going to help you much anyhow. At least 200C is required for modest bakes. The purpose of a bakeout is to get water off the walls and you have to overcome the heat of chemsorption.
A better way is to rig a leak of nitrogen from a liquid nitrogen container (open to atmosphere) so that you get flow in the viscous flow range. You have to throttle your vacuum pump so if you do not have a way to do this with your high vacuum pump, do it with your mechanical pump and throttle your leak. I think that if your run at about 0.1 Torr into a typical 1.5 inch vacuum line that you will be in the viscous range.
If you use clean stainless steel or copper tubing that is coiled and put your heat tape around that, you can heat the nitrogen gas going into your system for added benefit. Run this leak overnight and it will take out all the water off of the walls.
There is a commercially available system to do this and more. Contact Ron Vane at XEI.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Bruce Girrell [mailto:bigirrell-at-microlinetc.com] } Sent: Monday, March 26, 2001 10:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Bake out } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Greetings, } } I am very new to electron microscopy, so I ask your } indulgence in some basic } questions. } } I have an old JEOL SEM that has been sitting at atmospheric } pressure for } some time now and will need a thorough baking. It has a plain } old tungsten } wire filament, so the vacuum requirements aren't all that } stringent. The SEM } has rubber o-rings (I don't know how to tell if they are } Viton). I doubt } that I want to go above 100 oC. } } 1) What characteristics are important in choosing a heat tape } suitable for } baking the instrument? } } 2) Where would I find a suitable heat tape (my web searches } on heat tapes } got me lots of information about ice buildup on my roof, but } very little } about getting water out of an SEM)? } } 3) How do I determine how much to buy - how much column on } each side of the } tape can I assume to be heated properly by the tape - what is } an appropriate } center-to-center distance between adjacent wraps of tape? } } 4) Is there a more practical method than heat tapes for baking the } instrument? } } 5) What else may I be missing here? } } Thank you, } } Bruce Girrell } in snowy northern Michigan with a few more basic questions } waiting in the } wings } }
We have recently decommissioned a Fullam closed circuit TV imaging system, purchased in the late 80's-early 90's for use on a JEOL-1200EX. The system is in excellent condition, includes a side-port camera mount and detector, Dage MTI 70 camera, Nikkor lens, etc., and is available for donation to a school or university.
If interested, please respond directly to me and not to the list.
Regards, Bev Maleeff
GlaxoSmithKline Pharmaceuticals Safety Assessment 709 Swedeland Road M/C UE 0462 King of Prussia, PA 19406
I think a bigger question here is why Bruce thinks he needs a bake out? If he "heard" from someone that is what you have to do for a vacuum system then we should give him some easier advice.
First, we must all understand that most SEMs are not ultra high vacuum system that require baking. SEMs have specimen chamber with a typical vacuum between 10-5 to 10-6 Torr. High vacuum at best. An old JEOL SEM was never baked. Water vapor does not bother the electron beam or the secondary electrons at these pressures. I would clean the vacuum system with IPA, check and change as needed the pump oil, and try pumping down. Then I would fix any leaks.
Bruce, have you tried to pump it down? What make you think you should bake it? If oily dirt is a problem, then nitrogen purging like Scott Walck suggests might be a solution after you wipe down the chamber wall with IPA.
I have a long discussion of oil and hydrocarbon contamination and removal methods at www.SEMCLEAN.Com.
Ronald Vane XEI Scientific Notice: XEI makes anti-contamination system for SEMs.
Bruce have you tried pumping it down?
-----Original Message----- } From: Walck, Scott D. {walck-at-ppg.com} To: 'bigirrell-at-microlinetc.com' {bigirrell-at-microlinetc.com} Cc: 'Microscopy' {microscopy-at-sparc5.microscopy.com}
I appreciate the many responses that I have gotten already.
Several have questioned the need for a bake out. Well, I did say I was new to this...
The reasons I felt that the instrument should be baked are: 1) Wilbur Bigelow, in _Vacuum Methods in Electron Microscopy_ suggests that, short of pumping endlessly, baking is almost a necessity to remove adsorbed water. 2) The instrument has been sitting at atmospheric pressure for an untold period of time, at least several months. The E-T detector had been removed, leaving a nice 6" dia. hole in the system. God knows what else besides water vapor is in there. 3) The logbook that came with the unit recorded routine baking. Along with item 1), I concluded that baking was simply just a matter of course.
What I am hearing so far is that 1) for the relatively low vacuum required for this instrument, baking may not be necessary and 2) short of going to about 200C, baking may not be very effective.
Baking won't be necessary, you're biggest problem will be the o-ring seals. I assume this is a diffusion pump system.
O-ring seals have a tendency to dry out and shrink, if they've been in the system for some time, they will also have been shaped by the pressure of the flanges. The result will be poor mating with the flanges and lots of leaks.
You'll need to take each one out of the system and inspect it for drying and cracking. I do mean each one - if the system has been at atmosphere for a long time, you'll want to get every o-ring that seals the sample stage controls, vacuum valves and the gun translation mechanism, if so equipped.
If the o-rings are not too badly out of round and not cracking, then they can generally be reconditioned by cleaning them off with acetone and applying a very light layer of vacuum grease evenly on the surface. The grease is only to seal the surface, prevent drying and increase flexibility. I use acetone, although many will probably warn you not to, because it slightly swells the o-ring. When you're doing a general reconditioning like this, that helps the o-rings seal up right away, but it will still take a day or two at high vacuum to get them to fully seal.
This is also a good time to get in and clean the column and wipe down the sample and gun chambers. It's hard to say what will work here for you, as it depends largely on the pump oils that have been used in the past and the accumulated contamination from samples. Try some standard solvents like methanol, ethanol, acetone, etc. Hopefully one or a combination can remove some of the build-up.
The outgassing rates of adsorbed gasses at the vacuum levels you'll attain in the instrument are not really a problem. You'll want to leave the instrument in high vacuum for at least 24 hours. Over the first day or two, you will notice that the vacuum level slowly improves, primarily due to the o-rings being sucked into better contact with the flanges. Over this time, the system will also reach equilibrium with outgassing products.
On Monday, March 26, 2001 9:47 AM, Bruce Girrell [SMTP:bigirrell-at-microlinetc.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings, } } I am very new to electron microscopy, so I ask your indulgence in some basic } questions. } } I have an old JEOL SEM that has been sitting at atmospheric pressure for } some time now and will need a thorough baking. It has a plain old tungsten } wire filament, so the vacuum requirements aren't all that stringent. The SEM } has rubber o-rings (I don't know how to tell if they are Viton). I doubt } that I want to go above 100 oC. } } 1) What characteristics are important in choosing a heat tape suitable for } baking the instrument? } } 2) Where would I find a suitable heat tape (my web searches on heat tapes } got me lots of information about ice buildup on my roof, but very little } about getting water out of an SEM)? } } 3) How do I determine how much to buy - how much column on each side of the } tape can I assume to be heated properly by the tape - what is an appropriate } center-to-center distance between adjacent wraps of tape? } } 4) Is there a more practical method than heat tapes for baking the } instrument? } } 5) What else may I be missing here? } } Thank you, } } Bruce Girrell } in snowy northern Michigan with a few more basic questions waiting in the } wings } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
We are relatively new to atomic force microscopy and need some estimate of costs for routine maintenance. I don't mean expendible items but repairs.
For example, we have been facing some (in my opinion) rather high costs to repair hardware: broken scanner $3,900, electronic module $400 (twice). Are AFM's prone to such frequent, high repair costs?
Has anyone ever had to replace a scanner that "broke" for no apparent reason? BTW, none of these repairs were caused by operator error.
Just wondering.......
Thank you,
John
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
In conducting RGA studies of polymer outgassing inside hermetic electronic packages some years ago we found that hydrogen seems to have the effect of displacing moisture adsorbed onto metal surfaces. It was a problem that amounted to a "memory effect" within the RGA* for us but the effect can be used to advantage in displacing water. You might try Scott's suggestion substituting bottled, high purity hydrogen for the liquid nitrogen bleedoff. Obviously, hydrogen is flammable and requires caution when in use.
(* e.g. if a hermetic package punctured in the RGA sample compartment contained moisture, some of the released water vapor would adsorb onto the chamber wall. If the next hermetic package contained hydrogen, it would displace the moisture from the previous package causing it to be attributed to the second unit when, in fact, it was left over from the first. During pumping out of the sample compartment in between samples, the background level of moisture would be achieved even though it was lurking on the metal surface waiting to be liberated.)
John Twilley Conservation Scientist
Walck, Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would not recommend baking out that system if it has not been designed for it. The heat tapes will give you hot spots and other places may not take the heat well. If you are not going above 100C, it is not going to help you much anyhow. At least 200C is required for modest bakes. The purpose of a bakeout is to get water off the walls and you have to overcome the heat of chemsorption. } } A better way is to rig a leak of nitrogen from a liquid nitrogen container (open to atmosphere) so that you get flow in the viscous flow range. You have to throttle your vacuum pump so if you do not have a way to do this with your high vacuum pump, do it with your mechanical pump and throttle your leak. I think that if your run at about 0.1 Torr into a typical 1.5 inch vacuum line that you will be in the viscous range. } } If you use clean stainless steel or copper tubing that is coiled and put your heat tape around that, you can heat the nitrogen gas going into your system for added benefit. Run this leak overnight and it will take out all the water off of the walls. } } There is a commercially available system to do this and more. Contact Ron Vane at XEI. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } } } -----Original Message----- } } From: Bruce Girrell [mailto:bigirrell-at-microlinetc.com] } } Sent: Monday, March 26, 2001 10:47 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Bake out } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } } } } } -------------------------------------------------------------- } } ---------. } } } } } } Greetings, } } } } I am very new to electron microscopy, so I ask your } } indulgence in some basic } } questions. } } } } I have an old JEOL SEM that has been sitting at atmospheric } } pressure for } } some time now and will need a thorough baking. It has a plain } } old tungsten } } wire filament, so the vacuum requirements aren't all that } } stringent. The SEM } } has rubber o-rings (I don't know how to tell if they are } } Viton). I doubt } } that I want to go above 100 oC. } } } } 1) What characteristics are important in choosing a heat tape } } suitable for } } baking the instrument? } } } } 2) Where would I find a suitable heat tape (my web searches } } on heat tapes } } got me lots of information about ice buildup on my roof, but } } very little } } about getting water out of an SEM)? } } } } 3) How do I determine how much to buy - how much column on } } each side of the } } tape can I assume to be heated properly by the tape - what is } } an appropriate } } center-to-center distance between adjacent wraps of tape? } } } } 4) Is there a more practical method than heat tapes for baking the } } instrument? } } } } 5) What else may I be missing here? } } } } Thank you, } } } } Bruce Girrell } } in snowy northern Michigan with a few more basic questions } } waiting in the } } wings } } } } } } }
I am looking for a X-ray radiation detector used on TEMs. Currently we have a Geiger counter that is only sensitive to low energy X-ray. The purpose of the new detector is to sense high energy X-ray leakage from a 200 kV scope.
Any suggestion or clue will be greatly appreciated. Please contact off-line.
There's a whole chapter (#4) on support films in Hayat MA, Miller SE. 1990. Negative Staining. Pages 201-214 are on perforated films and discuss making of films with different size holes.
On Fri, 23 Mar 2001, Gordon Vrololjak wrote:
} X-Sieve: cmu-sieve 1.3 } Return-Path: {Microscopy-request-at-sparc5.microscopy.com} } Received: from jeter.acpub.duke.edu (jeter.acpub.duke.edu [152.3.233.10]) } by moorcock.acpub.duke.edu (8.9.3/8.9.3/Duke-5.0.0) with ESMTP id UAA15529 for {saram-at-moorcock.acpub.duke.edu} ; } Fri, 23 Mar 2001 20:14:59 -0500 (EST) } Received: from ultra5.microscopy.com ([206.69.208.10]) } by jeter.acpub.duke.edu (8.9.3/8.9.3/Duke-5.0.0) with ESMTP id UAA05887; } Fri, 23 Mar 2001 20:11:33 -0500 (EST) } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id RAA13779 } for dist-Microscopy; Fri, 23 Mar 2001 17:51:15 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id RAA13774 } for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 23 Mar 2001 17:50:45 -0600 (CST) } Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.175.19]) } by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id RAA13763 } for {microscopy-at-sparc5.microscopy.com} ; Fri, 23 Mar 2001 17:50:34 -0600 (CST) } Received: by nature.Berkeley.EDU (Postfix, from userid 7458) } id DFB298B64; Fri, 23 Mar 2001 15:48:36 -0800 (PST) } Received: from localhost (localhost [127.0.0.1]) } by nature.Berkeley.EDU (Postfix) with ESMTP id D3AB54DB35 } for {microscopy-at-sparc5.microscopy.com} ; Fri, 23 Mar 2001 15:48:36 -0800 (PST) } Date: Fri, 23 Mar 2001 15:48:36 -0800 (PST) } From: Gordon Vrololjak {gvrdolja-at-nature.Berkeley.EDU} } To: {microscopy-at-sparc5.microscopy.com} } Subject: ? how to make lacey carbon films? } Message-ID: {Pine.GSO.4.33.0103231546140.28537-100000-at-nature.Berkeley.EDU} } MIME-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Everyone, } I want to make my own lacey carbon films for cryo-tem and was wondering if } anyone had some good methods, experiences, insights, or references? } } I found references for lacey formvar, pure carbon films, and a few } others... But nothing explicitely for lacey carbon. } } All comments greatly appreciated, Thanks. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I would appreciate hearing how the experts out there prepare their microscope slides for measuring a PSF. We got a couple of prepared slides from Molecular Probes and were quite disappointed with the quality of the Blue and Green beads (the red ones were fine). This was somewhat surprising since they usually have great quality. We bought the beads and are making our own but I would be interested in any protocols or alternative sources for prepared slides. Thanks in advance, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
A W gun system doesn't need a particularly high vacuum to operate. But a better vacuum will extend the life of the filament dramatically. While I am not familiar with your JEOL instrument, I have worked with Amray 1610T and 1830 systems which had an ion pump so a LaB6 cathode could be used. Each system had separate guns for W and LaB6. I only ran W in the 1610T but only LaB6 in the 1830. In this respect, the ion pump had to be working and the vacuum had to be quite good.
When the gun chamber was opened, either for column liner cleaning, liner aperture change, anode aperture change or for cathode change and Wenhelt change, I baked out the gun chamber. If the ion pump indicated an inability to pump down quickly, I baked it out too. I never baked the chamber. Usually, when a system would sit around without being under vacuum, a long pumping after internal wipe down with methanol would bring the chamber vacuum back. If not, that is unchartered territory for me.
I baked out the gun chamber and the ion pump at 175F. All pumps (except ion) were on during the bake. I used flexible heating tape made by BH Thermal http://www.bhthermal.com and their control thermostat. The thermostat control is PN TS0991-550 and has a fluid bulb sensor. I used two silicone rubber extruded heating tapes, PN BS0101060L (313 Watts, 1" x 72") for gun and PN BS0201040L (418 Watts, 2" x 48") for pump. The first tape cost $95 and the second was $135 list. The controller was about $175.
The tape is wound tightly around the chamber or pump and taped down with high temperature fiberglass tape. This is PN PSAT36 (1/2" x 180ft) and costs $15 a roll. PN AAT260 (2" x 180ft) is also handy...$61 per roll. Wrapped around the heating tape is thick aluminum foil (cooking variety) and taped tight to the heating tape. The sensor bulb is taped down to the top of the gun assembly and should be enclosed by the aluminum foil. This makes it a set and forget deal for overnight baking.
The gaskets in my systems were Viton and copper. At 175F, I suspect rubber components would be OK. As far as I know, new replacements are Viton these days. But I can't say for certain if your system has rubber or Viton or even if replacements are Viton. Try heating a spare gasket or O-ring enclosed in aluminum foil in an oven at 175F and see if it is destroyed. That ought to tell the story. If the logs say that the system was baked, 175F ought to be safe.
The difference baking made was quite remarkable. If I did not bake, the ion pump would draw about 100-120uA and get worse as the cathode was on for much time. After baking, it idled at 50-60uA and would stay there for a long time.
The problem with a W-only system is that the gun chamber is typically connected to the specimen chamber via a stand pipe or some other vacuum connection. This means that the gun area is basically at the vacuum level of the chamber. Thus, my discussion of baking the gun chamber may be useless to you. But you could apply the same baking principles to the chamber and the gun chamber. I would not recommend at all baking or heating the column. This would surely destroy the scan coils.
If you have a turbo pump system, it is probably worth baking the system to purge the trapped moisture. If not, then it may not be of any value to bake. What kind of pumps does the system have?
Hope this is a good starting point for you.
gary g.
At 07:47 AM 3/26/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone know where one could get a microprocessor of the type that was used in the Polaron E5200 automatic coater. This was their first version of an automatic coater and the replacement part is no longer available via the manufacturer. Please respond to me off-line since this topic is probably not of interest to many followers of this list. If anyone is interested in the responses, let me know and I will send a summary of anything I learn.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Thanks to everyone who sent me their ideas for adhering sections to slides. We have had lots of suggestions and I will certainly post any successful results we have from the input,
As sometimes happens, some people find they can't get money to attend courses at the last minute while others don't hear about the courses until it is too late to apply.
Now may be your chance!
Several late cancellations at the UBC Live-Cell course mean three opportunities for those of you who have ten days free this June. In addition, the presence of more microscopy systems allows us to set up one more group-of-three.
We expect to have an international faculty of 15, and on-site equipment that includes 2-photon systems from Zeiss and Bio-Rad. There will also be single-photon confocals from Bio-Rad, Olympus, Nikon, Yokogawa/Solamere and Yokogawa/EG&G as well as deconvolution set-ups from Applied Precision Instruments, AutoQuant/Universal-Imaging, Improvision, Intelligent-Imaging- Innovations and Zeiss, all set up for "live-cell" work. In addition, there will be a laser tweezers/scissors from Cell-Robotics, microinjection from Eppendorf and sundry other delights including a fully equipped cell biology lab (with tireless technician!!) just for students.
These microscopy systems are not just to look at but to use for over 45 hours of organized 2D and 3D live-cell laboratory sessions, plus 10 hours of evening sessions for group live-cell projects.
Although the eight "Groups-of-3" have already been set up and have chosen their "individual projects," we are able to accept 3-2 more students as long as their backgrounds will fit into one of these groups, and we may even be able to set up one additional group.
which has the rest of the story, including a preliminary program.
Then fill out the Registration Form from the site to tell us about you, Fax it to Dr. Elaine Humphrey ASAP and we will see if we can fit you in. (I will be away April 1-14)
Hope that you can join us in Vancouver this June 18-28.
Jim Pawley, Organizer -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
As pointed out by many listers, it may not be necessary to bake your instrument. Since the baking part has been well covered, I'll just add a very effective way to tell Viton o-rings from rubber ones. If you can find some Freon TF just put some in a beaker and drop in the questionable o-ring. If the o-ring floats, it is rubber and if it sinks, it is Viton.
Good luck, Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone 512/282-5507 FAX 512/280-0702
Sustaining Member - MICROSCOPY SOCIETY OF AMERICA
-----Original Message----- } From: Bruce Girrell {bigirrell-at-microlinetc.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Greetings, I am taking overlapping video images of a growing root through a horizontal light microscope. For analysis, I need to put the images back in register. I have tried incorportating various high contrasty things into the background but this introduces various problems ranging from death to the root to requiring a large overlap between successive images. As an alternative, I thought of measuring the displacement of the stage. A typical displacement between images is around 300 microns and I need to measure this displacement to within around 1 micron. I only need to work in one dimension, which can be parallel to one of the edges of the stage. I have in mind something like a "giant" afm detection set up, with a laser bouncing off the edege of stage and picked up by a detector. But this is just wild imagination on my part. Anyone out there have any *knowledge* about how this might be accomplished?
Thanks in advance, Tobias -- _ ____ __ ____ Tobias I. Baskin / \ / / \ / \ \ 109 Tucker Hall / / / / \ \ \ Biological Sciences /_ / __ /__ \ \ \__ University of Missouri / / / \ \ \ Columbia, MO USA / / / \ \ \ 65211-7400 / / ___ / \ \__/ \ ____ voice: 573-882-0173 fax: 573-882-0123
I was a student intern with Dr. Judi Walton when she was first using lead aspartate in 1977. I don't remember if we used it with tissue, but I know we used it with cell culture monolayer samples for TEM. We used it without poststaining. She developed this as Walton's Lead Aspartate. I don't know if it is still on the market....
Barbara L. Plowman Univ. of the Pacific School of Dentistry 2155 Webster Rm 632 San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu
Thank you for the answers to my inquiry regarding 3D reconstruction of TEM serial sections and / or EM tomography. They were very useful and we have a much better idea of how to proceed with our project. I have, at the request of a couple of list subscribers, summarized the answers in a Word file and sent the file to them. It is available to anyone who emails me and asks - it is kind of long, but there is good advice and contact information included for anyone who is beginning in this area.
Again - Thanks to all who helped. I wouldn't know how to cover the bases for those who depend on me without this list! Thanks to Nestor, too!
Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Dear Readers, I have a Nikon 63x oil objective that is used on an inverted scope. It is a multi user facility and I just noticed today that oil has seeped into the objective making it almost useless to use.
(i) Should I tackle the job of cleaning this lens myself or (ii) Does anyone know of a facility that would clean this lens at a reasonable price?
I used to have silicon around the top of the lens to prevent this problem but it came off and I forgot to replace it.
Thanks for any information on this topic.
Sean M. Ward Associate Professor Department of Physiology and Cell Biology University of Nevada School of Medicine Manville Medical Sciences Building Reno NV 89557 Tel: (775) 784-6061 Fax:(775) 784-6903
I've been asked to look at the microscopic distribution of a mineral oil used in plant sprays on a plant surface. The spray is dispersed in water at 1-2% so the final volumes on the surface can be quite low. Most of the techniques I have seen are concerned with the total distribution of the spray and use either fine particles or dyes in the aqueous phase and do not track the oil. I have had only limited success with the natural autofluorescence of the oil, cathodoluminescence in the SEM and with common chromatic dyes such as oil red. Visualization is just possible if we deposit pure oil on the surface but not the suspension. Has anyone any suggestions??
Thanks in advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
Yes, an interferometer could be used to measure the stage movement, but it's an expensive option. If you truly need 1 micron placement, there are few methods available. If you can accept somewhat less resolution, a machinist's dial micrometer with a stand can give you 1/10,000 of an inch measurement rather inexpensively, but with limited travel. You didn't mention the overall length of your samples.
Being based at a university, try your local campus machine shop. Machinists tend to covet those precision instruments, but perhaps you could borrow one for a short time (just don't sign in blood).
On Tuesday, March 27, 2001 2:33 PM, Tobias Baskin [SMTP:BaskinT-at-missouri.edu] wrote: } } Greetings, } I am taking overlapping video images of a growing root } through a horizontal light microscope. For analysis, I need to put } the images back in register. I have tried incorportating various high } contrasty things into the background but this introduces various } problems ranging from death to the root to requiring a large overlap } between successive images. As an alternative, I thought of measuring } the displacement of the stage. A typical displacement between images } is around 300 microns and I need to measure this displacement to } within around 1 micron. I only need to work in one dimension, which } can be parallel to one of the edges of the stage. I have in mind } something like a "giant" afm detection set up, with a laser bouncing } off the edege of stage and picked up by a detector. But this is just } wild imagination on my part. Anyone out there have any *knowledge* } about how this might be accomplished? } } Thanks in advance, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123 } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Dear collegues has anyone out there has any experiences with imaging bacteriophages in a FESEM and does anyone know of some published pictures. Thank's in advance. Manfred
Ian I have never tried this, but one possibility would be to add a fluorochrome to the system. Nile red is an oil-soluble fluorochrome, only sparingly soluble in aqueous phase, strongly fluorescent only when it partitions into an apolar environment but non-fluorescent in aqueous phase. It should be possible to image Nile-red labelled oil droplet footprints by fluorescence LM with FITC-type filter systems and possibly also by cathodoluminescence in SEM. If you are working on leaves the red autofluorescence of the chloroplasts should be filter out with a green barrier filter. The Leica GFP Plant filter sets may therefore be suitable. see Greenspan, P., Mayer,E.P. and Fowler,S.D. (1985) Nile red: a selective fluorescent stain for intracellular lipid droplets. Journal of Cell Biology 100, p965-973.
Hope this helps Chris ----- Original Message ----- } From: "IAN HALLETT" {ihallett-at-hortresearch.co.nz} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 28, 2001 5:04 AM
1 micron is a pretty demanding tolerance. Do you really need it to be this good? Stages used to be provided with x and y vernier scales, but today a stepper motor stage drive is often used controlled by a computer which counts the steps. Mitutoyo make electronic measuring devices accurate to about 5 or 10 microns which are used in electronic calipers. Similar modules are available for mounting on engineering x,y stages, for example of milling machines. Electronic supplies companies like Farnell, RS Components and others supply these.
----- Original Message ----- } From: "Tobias Baskin" {BaskinT-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, March 27, 2001 9:33 PM
The Department of Engineering Metals, Chalmers University of Technology (Göteborg, Sweden) is seeking:
DOCTORAL STUDENT Microstructure and Chemistry of Nanostructured Materials using Transmission Electron Microscopy
Any technological application of nanostructured materials is only possible if their properties and their microstructure are well understood so that they can be manufactured reproducibly. Concentrating on Ni- and Co-based materials analytical transmission electron microscopy will be applied to characterize the microstructure of the material as well as to monitor the microstructural evolution upon heating. The work will be performed in close collaboration with colleagues at the University of Toronto, Canada. The experiments will be carried out on a Zeiss 912 Omega with inbuilt imaging energy filter, however, also a Philips CM 200 FEG equipped with EDS and Gatan imaging filter (GIF) is accessible. A special heating holder is available to perform the in-situ heating experiments.
The appointment applies to full time for a maximum period of five years. In addition to the research activities departmental duties, i.e. a limited amount of educational, research and/or administrative work up to 20% of full time has to be performed. The initial monthly salary is SEK 18.000, however depending on the academic degree. The applicant must have an academic degree (MSc, BSc or equivalent) in materials science/materials engineering, physics or a similar subject area. Experience in (analytical) transmission electron microscopy is a merit. The position is available immediately.
For further information about the position, please contact: Prof. Uta Klement Chalmers University of Technology, Department of Engineering Metals Tel: +46 31-772 1264, Fax: +46 31-772 1262 E-mail: Klement-at-em.chalmers.se
A written application including the reference number 23/2001, with (attested) personal record, brief description of research interests, name and e-mail addresses to three reference persons, possible day for taking possession of the position, and the documents that you want to refer to should have arrived at the Registrar, Chalmers University of Technology, SE-412 96 Göteborg, Sweden no later than April 27, 2001.
I can't visualize exactly how you are doing this, but I assume you are using reflected rather than transmitted light.
In the dim and distant past I have used a 'traveling microscope' which consists of a lens system with a cross-hair which can be moved along a metal rail (at least 200mms long). The rail had divisions marked off accurately in mms and a sliding Vernier scale so I would guess that it would be accurate to 10um or maybe better. I suppose that it may be possible to attach a camera system instead of the lens but it would be best to try and borrow one to find out. I assume that a stage micrometre (slide marked off in fractions of 1mm or 10mm) could not be adapted to your purposes but it might be a useful way of calibrating any system you adopt.
Malcolm
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Tobias Baskin wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings, } I am taking overlapping video images of a growing root } through a horizontal light microscope. For analysis, I need to put } the images back in register. I have tried incorportating various high } contrasty things into the background but this introduces various } problems ranging from death to the root to requiring a large overlap } between successive images. As an alternative, I thought of measuring } the displacement of the stage. A typical displacement between images } is around 300 microns and I need to measure this displacement to } within around 1 micron. I only need to work in one dimension, which } can be parallel to one of the edges of the stage. I have in mind } something like a "giant" afm detection set up, with a laser bouncing } off the edege of stage and picked up by a detector. But this is just } wild imagination on my part. Anyone out there have any *knowledge* } about how this might be accomplished? } } Thanks in advance, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123
Hi, All, I take some diffraction contrast pctures. As the thickness of the samples changes sharply the brightness of the images is not uniform. Does someone know any software to adjust the brightness for the whole image? Thanks inadvance. Best regards. Feng ********************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
A good tool maker should be able to make you someting the will do an order of magnitued beter than 1/10,0000 by using a standard micrometer head and setting to drive a wedge at about 10 degrees. I don't remember the exact angle but i used it to get .00001 on a lathe wiht 0001 devisions. I dont expect you would get all the precision but you sould get .00022 to .00004
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "Allen R. Sampson" {ars-at-sem.com} To: "'Tobias Baskin'" {BaskinT-at-missouri.edu} ; {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 28, 2001 12:03 AM
Hi folks, If anyone has experience with and opinions about the product Auto- Montage by Syncroscopy, please contact me or Mark Farmer off- listserve. Thanks in advance,
John Shields EM Lab University of GA Athens jshields-at-cb.uga.edu
Hi Sean, We had the same problem with one of our objectives on the Nikon inverted. The lens should not allow oil in if it is sealed correctly, even without the "seal" you provided. Your lens is either older and the seal is worn, or you've been cleaning the oil off with a harsh solvent (I've seen some people using xylene regularly). Somewhere in the back of my mind (a dusty and unreliable source) I think silcon is not good for the seal. Another solution is to take finger cots or make them by cutting the finger tips from gloves and pull them down over the lens if you are still unsure of your seal. We ended up sending the lens to Nikon for repair. I seem to remember it cost around $200 or so to have them clean it and replace the seal, but you can call to get a quote. good luck
On 27 Mar 2001, at 17:39, Sean Ward wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Dear Readers, } I have a Nikon 63x oil objective that is used on an inverted scope. } It is a } multi user facility and I just noticed today that oil has seeped into } the objective making it almost useless to use. } } (i) Should I tackle the job of cleaning this lens myself or (ii) Does } anyone know of a facility that would clean this lens at a reasonable } price? } } I used to have silicon around the top of the lens to prevent this } problem but it came off and I forgot to replace it. } } Thanks for any information on this topic. } } } Sean M. Ward } Associate Professor } Department of Physiology and Cell Biology } University of Nevada School of Medicine } Manville Medical Sciences Building } Reno NV 89557 } Tel: (775) 784-6061 } Fax:(775) 784-6903 } }
John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
Hi Tobias, What you suggest may not be worth the hassle. Potential error introduced by temp. variations on the stage would be problematic. Is it not possible to include an internal standard, for example an embedded stage micrometer, ruler, calibrated cover plate for the incubation vessel,... Ramin
-----Original Message----- } From: Tobias Baskin [mailto:BaskinT-at-missouri.edu] Sent: Tuesday, March 27, 2001 3:33 PM To: Microscopy-at-sparc5.microscopy.com
Greetings, I am taking overlapping video images of a growing root through a horizontal light microscope. For analysis, I need to put the images back in register. I have tried incorportating various high contrasty things into the background but this introduces various problems ranging from death to the root to requiring a large overlap between successive images. As an alternative, I thought of measuring the displacement of the stage. A typical displacement between images is around 300 microns and I need to measure this displacement to within around 1 micron. I only need to work in one dimension, which can be parallel to one of the edges of the stage. I have in mind something like a "giant" afm detection set up, with a laser bouncing off the edege of stage and picked up by a detector. But this is just wild imagination on my part. Anyone out there have any *knowledge* about how this might be accomplished?
Thanks in advance, Tobias -- _ ____ __ ____ Tobias I. Baskin / \ / / \ / \ \ 109 Tucker Hall / / / / \ \ \ Biological Sciences /_ / __ /__ \ \ \__ University of Missouri / / / \ \ \ Columbia, MO USA / / / \ \ \ 65211-7400 / / ___ / \ \__/ \ ____ voice: 573-882-0173 fax: 573-882-0123
I have heard mention of replicating surfaces with carbon for analysis w/ EM, it seems like it may be history now, but I was wondering if someone could "clue me in" with when it was required.
At 5:39 PM -0800 3/27/01, Sean Ward wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You have my deepest sympathies! We had the same experience with a Zeiss lens on a multi-user Axiovert a few years ago. I would NOT recommend trying this on your own. High power lenses are complexes of many lens elements and, in my humble opinion, should only be disassembled by those who know what they are doing. We ended up sending our lens back to Germany. 6 weeks and many $$$ later, it was like new.
Ask you Nikon dealer what s/he would suggest.
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Is this correct? There isn't such a tight seal at the telescoping part of the objective barrel - this (in my experience) is where oil can get in unless a lens condom is used - or have I been woefully misled?
Tamara
On Wed, 28 Mar 2001 jshields-at-cb.uga.edu-at-sparc5.microscopy.com wrote:
} } Hi Sean, } We had the same problem with one of our objectives on the Nikon } inverted. The lens should not allow oil in if it is sealed correctly, } even without the "seal" you provided. Your lens is either older and } the seal is worn, or you've been cleaning the oil off with a harsh } solvent (I've seen some people using xylene regularly). Somewhere } in the back of my mind (a dusty and unreliable source) I think } silcon is not good for the seal. Another solution is to take finger } cots or make them by cutting the finger tips from gloves and pull } them down over the lens if you are still unsure of your seal. } We ended up sending the lens to Nikon for repair. I seem to } remember it cost around $200 or so to have them clean it and } replace the seal, but you can call to get a quote. } good luck }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
You might try an ordinary dial gauge, a device widely used for measuring small movements in machine shops. Check with your friendly machinist, or try your local hardware store. Dial gauges will easily measure 0.0005" (0.01 mm), and are simple, inexpensive, and easy to use. I am sure you can order one from Small Parts Co. I don't happen to have their address here, but you can probably find them on the internet.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Prior to the introduction of techniques for preparing and examining thin metal specimens by transmission electron microscopy, which came into general use in the late 1950s, virtually all electron microscopic studies of non-biological materials was done by examining exposed surfaces (treated in various ways to reveal the characteristics of the internal structure) using replical techniques. Early on, these replicas were made of various polymeric materials (e.g. formvar, colloidion, etc.). There were various shortcomings of these replicas, mainly their resolution was limited to something of the order of 50 Angstroms. In 1954 D. E. Bradley introduced a convenient method for depositing very thin films of carbon. These films were proven to be much better than the polymer films for producing surface replicas. They were stronger, they were electrically conductive, they could be made much thinner, but most of all could provide resolution below 20 Angstroms. Therefore they quickly became the most widely used type of replica film. You can find a rather complete discussion of the many replica techniques that were in use back then in the book 'Techniques for Electron Microscopy', D. E. Kay, Editor, Blackwell Scientific Pub., 1965. These techniques are hardly ever mentioned now-a-days, because there are so many newer techniques now available for producing thin specimens for direct examination.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Listers, I received this inquiry recently. Can anybody help this person? Hi. I am a biology student and I have to do an assignment on electron or light microscopy an it's impact on society. For example, in forensic science or medical research. If you have any information that would help with this, me please email it to me as soon as possible. It would be greatly appreciated. Thankyou, Nicole.
Nicole Forzan {phindola16-at-hotmail.com} Thanks, Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
} Date: Wed, 28 Mar 2001 13:20:14 -0800 } To: "Smartech" {smartech-at-javanet.com} } From: Mary Mager {mager-at-interchange.ubc.ca} } Subject: Re: EM, Why did they replicate surfaces w/ carbon? } Cc: Microscopy } } Dear Ric, } We routinely do carbon replicas of steel surfaces to study the very fine precipitates by TEM, free of the steel matrix. The method is: etch the steel lightly, carbon coat in the evaporator, score the carbon coat into 3 mm. squares with a razor blade, place the steel in the etchant until the carbon squares float off. Scoop up the floating carbon squares with a TEM grid. } For SEM of surfaces that are too big to go into the SEM chamber or that the cops won't let you take away, a cellulose acetate replica is used. It also removes surface material, allowing you to analyse it free of the matrix and cleaning the matrix. It is often used for fractures. The cellulose acetate film is softened in acetone and the sample surface is wetted with acetone. The acetate is placed on the acetone-wetted surface and left for about 0.5 hour to solidify. It conforms exactly to the surface, so you can study the fracture surface without putting the sample in the SEM, after coating the acetate piece to make it conductive. } That is very brief, you can contact me for more information. } At 10:17 AM 3/28/01 -0500, you wrote: } } } I have heard mention of replicating surfaces with carbon for analysis w/ EM, } } it seems like it may be history now, but I was wondering if someone could } } "clue me in" with when it was required. } } } } Thanks } } } } Ric } } } } SMARTech } } 860-491-3299 } } www.semguy.com } } 19 Cornwall Drive } } Goshen CT 06756 } } } Regards, } Mary } } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Can someone tell me what the mains supply voltage is in Sweden?
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
----- Original Message ----- } From: "Wil Bigelow" {bigelow-at-engin.umich.edu} } You might try an ordinary dial gauge, a device widely used for measuring } small movements in machine shops. Check with your friendly machinist, or } try your local hardware store. Dial gauges will easily measure 0.0005" } (0.01 mm), and are simple, inexpensive, and easy to use. I am sure you can } order one from Small Parts Co. I don't happen to have their address here, } but you can probably find them on the internet. } Wil,
Thanks for mentioning this it solves a problem I have on how to tell how much I move the stage for some experments I am doing trying to enhance the quality of video images by stacking multiple images. I need to get a little shift of the view for better averageing. I never thought using the test indicator laying 6 feet from the scope.
Looking through my machine tool catalog what you want if you go this route is called a test indicator. It is an order of magnitude more accrute than most dial indicators. They can be purchased with calimed accurcies down to 0.00012 or .003 mm. They are avalible in either mecanical dial or digital and range in cost from $100 to $500USD. They are built to be used in machine shops and should give very long service in a labrotory. The accurcy ranges from .001 to .00012 over the price range.
A lever arangement could be built to extend the resolution. It would require careful work to retain the accurcy.
If you need help in finding a vendor or how one would work I have spent countless hours using one.
good luck Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Panel components, Inc. info-at-panelcomponents.com has a great catalog with info on power,plugs, receptacles, etc. for most countries.
I have no $ interest...... but have purchased items from them.
Don Marshall
} From Microscopy-request-at-sparc5.microscopy.com Wed Mar 28 21:07:09 2001 From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } Organization: Dept of Geology, Univ of Auckland } To: Microscopy-at-sparc5.microscopy.com } Date: Thu, 29 Mar 2001 14:07:04 GMT+1200
} Hi } } Can someone tell me what the mains supply voltage is in Sweden? } } thanks } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to dmrelion-at-aol.com. Thank you.)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all those who responded to my question regarding enrobing of cells in agarose. Bruce Bruce Cutler Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id MAA03114 for dist-Microscopy; Thu, 29 Mar 2001 12:22:45 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id MAA03110 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 29 Mar 2001 12:22:14 -0600 (CST) Received: from srvr20.engin.umich.edu (srvr20.engin.umich.edu [141.213.75.22]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id MAA03103 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 29 Mar 2001 12:22:03 -0600 (CST) Received: from [141.213.21.161] ([141.213.21.161]) by srvr20.engin.umich.edu (8.9.3/8.9.1) with ESMTP id NAA02225; Thu, 29 Mar 2001 13:19:58 -0500 (EST) User-Agent: Microsoft-Entourage/9.0.2509
Let me actually try and answer your question.
In keeping with most of the rest of Europe I think it is 220/240.
On 3/29/01 8:47 AM, "donald j marshall" {dmrelion-at-world.std.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ritch, } } } Panel components, Inc. info-at-panelcomponents.com has a great } catalog with info on power,plugs, receptacles, etc. for most countries. } } I have no $ interest...... but have purchased items from them. } } } } Don Marshall } } } } } From Microscopy-request-at-sparc5.microscopy.com Wed Mar 28 21:07:09 2001 } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } } Organization: Dept of Geology, Univ of Auckland } } To: Microscopy-at-sparc5.microscopy.com } } Date: Thu, 29 Mar 2001 14:07:04 GMT+1200 } } } } Hi } } } } Can someone tell me what the mains supply voltage is in Sweden? } } } } thanks } } } } rtch } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } Department of Geology Fax : 64 9 3737435 } } The University of Auckland email : r.sims-at-auckland.ac.nz } } Private Bag 92019 } } Auckland } } New Zealand } } } } Donald J. Marshall } Relion Industries } P.O. Box 12 } Bedford, MA 01730 } Ph: 781-275-4695 } FAX: 781-271-0252 } email dmrelion-at-world.std.com } } Cathodoluminescence, mass spectroscopy, electron beam technology } } } "A weed is a flower out of place." } } (Please note: Do not send email with attachments to this address. Instead, } send it to dmrelion-at-aol.com. Thank you.) } }
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
} From http://www.sverigeturism.se/smorgasbord/smorgasbord/service/electricity.html:
The voltage is 230V in Sweden. All new buildings erected from January 1, 1994, will be in accord with EU standards. That is, all sockets are earthed. In every public milieu there will only be earthed sockets. In all other buildings erected before the date of January 1, 1994, earthed sockets will be found in bathrooms, as well as in kitchens. Otherwise there are simple sockets. In case one needs an adapter it can be acquired in an electricity store. There are no transformers for 130 volt to 230 volt to be found.
"John F. Mansfield" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Let me actually try and answer your question. } } In keeping with most of the rest of Europe I think it is 220/240. } } On 3/29/01 8:47 AM, "donald j marshall" {dmrelion-at-world.std.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } ritch, } } } } } } Panel components, Inc. info-at-panelcomponents.com has a great } } catalog with info on power,plugs, receptacles, etc. for most countries. } } } } I have no $ interest...... but have purchased items from them. } } } } } } } } Don Marshall } } } } } } } } } From Microscopy-request-at-sparc5.microscopy.com Wed Mar 28 21:07:09 2001 } } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } } } Organization: Dept of Geology, Univ of Auckland } } } To: Microscopy-at-sparc5.microscopy.com } } } Date: Thu, 29 Mar 2001 14:07:04 GMT+1200 } } } } } } } Hi } } } } } } Can someone tell me what the mains supply voltage is in Sweden? } } } } } } thanks } } } } } } rtch } } } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } } } Department of Geology Fax : 64 9 3737435 } } } The University of Auckland email : r.sims-at-auckland.ac.nz } } } Private Bag 92019 } } } Auckland } } } New Zealand } } } } } } } Donald J. Marshall } } Relion Industries } } P.O. Box 12 } } Bedford, MA 01730 } } Ph: 781-275-4695 } } FAX: 781-271-0252 } } email dmrelion-at-world.std.com } } } } Cathodoluminescence, mass spectroscopy, electron beam technology } } } } } } "A weed is a flower out of place." } } } } (Please note: Do not send email with attachments to this address. Instead, } } send it to dmrelion-at-aol.com. Thank you.) } } } } } } -- } } Dr. John Mansfield CPhys MInstP } North Campus Electron Microbeam Analysis Laboratory } 417 SRB, University of Michigan } 2455 Hayward, Ann Arbor MI 48109-2143 } Phone: (734) 936-3352 FAX (734) 763-2282 } Cellular Phone: (734) 358-7555 } (Leaving a phone message at 936-3352 is preferable to 358-7555) } Email: jfmjfm-at-engin.umich.edu } URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html } Location: Lat. 42° 16' 48" Long. 83° 43' 48"
-- Henrik Kaker, Ph.D. Metal Ravne d.o.o. SEM-EDS Lab Koroska cesta 14, 2390 Ravne Slovenia Phone: +386 602 21 131 Fax: +386 602 20 436 http://www.kaker.com
I would like to know what computer programs are out there for taking serial sections and creating a 3 D image. Can any one help? Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
{html} {font face="Times, Times"} Dear Listers, {br} {br} Immunogold Workshop Anouncement {br} {br} The Electron Microscopy Facility for the Life Sciences, a shared technology laboratory in the Life Sciences Consortium at Penn State University is hosting a three-day workshop on immunogold techniques from May 21-23, 2001. Dr. Jan Luenissen from the Aurion Immunogold Reagent & {br} Accessories, an internationally known expert in the field, will be the instructor for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The following is the workshop main curriculum. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-727-8692) or email (hyi-at-emory.edu). {br} {br} {br} MAIN CURRICULUM {br} {br} The properties of gold particles and their protein conjugates. {br} Theories underlying immunogold labeling protocols. {br} Silver enhancement of gold particles {br} Immunogold labeling on a variety of sample preparations for LM. {br} Immunogold labeling for EM {br} a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. {br} b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmalll gold conjugates. {br} Pre- and post-embedding double immunogold labeling. {br} Background minimization in immunogold labeling {br} Signal amplification in immunogold labeling. {br} {br} {br} Rosemary Walsh, Manager {br} The Electron Microscope Facility for the Life Sciences {br} A Shared Technology in The Life Sciences Consortium {br} 1 South Frear Lab {br} Penn State University {br} University Park, PA 16802 {br} (815) 865-0212 {br} {a href="http://www.lsc.psu.edu/stf/em/home.html" eudora="autourl"} http://www.lsc.psu.edu/stf/em/home.html {/a} {br} {br} {br} {br} {br} {/font} {/html}
There exists a homepage of 3-Dimensional Microscopy Labs (http://3dem.sdsc.edu/) in which are listed various packages for 3D-EM Image Analysis (including reconstruction of serial secions). Additionally a mailing list exists.
BTW: A nice animation can be found in a Web-Page of the University of Utrecht (The Netherlands): http://emsaserv.bio.uu.nl/3dem/ANIMATED_INTRODUCTION/animated_introduction_1.html
I hope this helps.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Herbststrasse 7 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: http://www.tvips.com/ Email: ingo.daberkow-at-tvips.com
-------- Original Message --------
Dear Listers:
I would like to know what computer programs are out there for taking serial sections and creating a 3 D image. Can any one help? Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Dear collegues, I need to perform an autoradiographic study in electron microscopy. From some papers in the literature it seems usuful to employ a phenidon developer and a gold latensification solution; since I did not find any commercial product available, I need to prepare it in my lab. I would be very grateful to you if you can provide me with some information about the composition and detailed method of preparation and storage of the different solutions. Thank you very much. Maria Meli
I am pleased to announce the completely revised and updated edition of "Microscopy & Imaging Resources on the WWW" is now available at a new URL. The goal of this site has always been to provide resources for University students, staff and faculty who want to learn more about microscopy and imaging. The site includes K-12 educational links, information on Light Microscopy, Histology, Confocal Microscopy, Fluorescence Techniques, Electron Microscopy, and Digital Imaging.
http://swehsc.pharmacy.arizona.edu/exppath/
"Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest Environmental Health Sciences Center, which is funded by the National Institute of Environmental Health Sciences (NIEHS is one of the National Institutes of Health, part of the U.S. Department of Health and Human Services).
Yours, Doug Cromey Manager, Experimental Pathology Core, SWEHSC .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : :...................................................................: http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
Listerservers, I have heard that there are a set of fonts called 'Fraser fonts' with over bars etc. that are useful for indices. Does it exist. Where can such a font be obtained?
Thanks in advance, Steven
-- Dr. Steven Celotto Department of Engineering Materials Science & Engineering University of Liverpool Brownlow Hill Liverpool L69 3BX UNITED KINGDOM
ok taking a little survey. i am in a diagnostic EM lab. we mount out sections on formvar coated slotted grids, so he can shoot pics of the whole glomerlus. ok my question. how may of you out there doing diagnostis EM on renals do this? john
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} ok taking a little survey. i am in a diagnostic EM lab. we mount out sections {BR} on formvar coated slotted grids, so he can shoot pics of the whole glomerlus. {BR} ok my question. how may of you out there doing diagnostis EM on renals do {BR} this? {BR} john {/FONT} {/HTML}
We have just acquired a used microscope made by ISI Inc. (their model alpha - 1980). Would anyone please have information on how we can obtain spare parts, filaments etc., for this machine. Many thanks for your help...
John: I did renal biopsies by E-M for 12 years and knew the other two people in town who were also doing diagnostic E-M of renal biopsies. We all used grids with grid bars ( I always used naked grids). I chose grids with slender bars for all my work to maximize viewing without using slotted grids. We did not view "whole" glomeruli slices (quite low power I would think). I don't feel diagnoses were compromised versus using slotted grids. Since several sections are mounted on the 200 mesh grids, one should be able to view areas of interest just fine.
} } } {"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com} 03/30 11:20 AM } } } ok taking a little survey. i am in a diagnostic EM lab. we mount out sections on formvar coated slotted grids, so he can shoot pics of the whole glomerlus. ok my question. how may of you out there doing diagnostis EM on renals do this? john
will add this link to the upcoming SMECC website. thanks Ed Sharpe
} Subj: Web site announcement } Date: 3/30/01 11:16:39 AM US Mountain Standard Time } From: Cromey-at-Arizona.edu (Doug Cromey) } To: microscopy-at-sparc5.microscopy.com, confocal-at-listserv.acsu.buffalo.edu } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } } I am pleased to announce the completely revised and updated edition of } "Microscopy & Imaging Resources on the WWW" is now available at a new } URL. The goal of this site has always been to provide resources for } University students, staff and faculty who want to learn more about } microscopy and imaging. The site includes K-12 educational links, } information on Light Microscopy, Histology, Confocal Microscopy, } Fluorescence Techniques, Electron Microscopy, and Digital Imaging. } } http://swehsc.pharmacy.arizona.edu/exppath/ } } "Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest } Environmental Health Sciences Center, which is funded by the National } Institute of Environmental Health Sciences (NIEHS is one of the National } Institutes of Health, part of the U.S. Department of Health and Human } Services). } } Yours, } Doug Cromey } Manager, Experimental Pathology Core, SWEHSC } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : } :...................................................................: } http://swehsc.pharmacy.arizona.edu/exppath/ } Home of: "Microscopy and Imaging Resources on the WWW" } } } } } ----------------------- Headers --------- }
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} will add this link to the upcoming SMECC website. {BR} thanks Ed Sharpe {BR} {BR} {BR} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} Subj: {B} Web site announcement {/B} {BR} Date: 3/30/01 11:16:39 AM US Mountain Standard Time {BR} {I} From: Cromey-at-Arizona.edu (Doug Cromey) {BR} To: microscopy-at-sparc5.microscopy.com, confocal-at-listserv.acsu.buffalo.edu {BR} {/I} {BR} {BR} {BR} {BR} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} -----------------------------------------------------------------------. {BR} {BR} {BR} Dear Colleagues, {BR} {BR} I am pleased to announce the completely revised and updated edition of {BR} "Microscopy & Imaging Resources on the WWW" is now available at a new {BR} URL. The goal of this site has always been to provide resources for {BR} University students, staff and faculty who want to learn more about {BR} microscopy and imaging. The site includes K-12 educational links, {BR} information on Light Microscopy, Histology, Confocal Microscopy, {BR} Fluorescence Techniques, Electron Microscopy, and Digital Imaging. {BR} {BR} http://swehsc.pharmacy.arizona.edu/exppath/ {BR} {BR} "Microscopy & Imaging Resources on the WWW" is an outreach of the Southwest {BR} Environmental Health Sciences Center, which is funded by the National {BR} Institute of Environmental Health Sciences (NIEHS is one of the National {BR} Institutes of Health, part of the U.S. Department of Health and Human {BR} Services). {BR} {BR} Yours, {BR} Doug Cromey {BR} Manager, Experimental Pathology Core, SWEHSC {BR} .................................................................... {BR} : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : {BR} : Research Specialist, Principal University of Arizona : {BR} : (office: AHSC 4212A) P.O. Box 245044 : {BR} : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : {BR} : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : {BR} :...................................................................: {BR} http://swehsc.pharmacy.arizona.edu/exppath/ {BR} Home of: "Microscopy and Imaging Resources on the WWW" {BR} {BR} {BR} {/XMP} {/FONT} {FONT COLOR="#0f0f0f" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {BR} ----------------------- Headers --------- {BR} {/BLOCKQUOTE} {BR} {/FONT} {/FONT} {FONT COLOR="#000000" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {/FONT} {/HTML}
ISI became Topcon, and then dropped out of the American market directly awhile ago. Any of the SEM supply houses can provide supplies or you can address your requests to Aspex, which was R. J. Lee until recently. They currently represent the Topcon lines in North America and can be found at http://www.rjleeinst.com/. Confused yet? It gets better. Good luck finding anything about ISI or Topcon products on their website. Try the phone lines.
Frankly, I hate having to trace the providence of this and other companies that give up well known brand names for reasons known only to them. ISI made inroads in this country primarily a s a low-bidder, thus many instruments were originally in government labs. Had they made some attempt to produce a product for the middle of the pack they may have done better and may still be a player.
I'm not saying that they didn't produce a decent instrument - I still have many that perform quite well. But they cut their own throats by playing to an audience that has suffered devastating budgetary cutbacks in an era when other segments were willing to pay vastly more for instruments that offer little basic operational improvements.
On Friday, March 30, 2001 2:53 PM, Rishi Raj [SMTP:Rishi.Raj-at-Colorado.edu] wrote: } ------------------------------------------------------------------------ } } We have just acquired a used microscope made by ISI Inc. (their model } alpha - 1980). Would anyone please have information on how we can obtain } spare parts, filaments etc., for this machine. Many thanks for your } help... } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
-----Original Message----- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, March 30, 2001 2:20 PM To: microscopy-at-sparc5.microscopy.com
-----Original Message----- } From: Garrison, Becky Sent: Saturday, March 31, 2001 8:33 AM To: '"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com'; microscopy-at-sparc5.microscopy.com
John: We do around 500 renal biopsies per year and all the sections are mounted on 200 mesh uncoated copper grids. We have an 8 year old Hitachi 7100 and use 60kv. The majority of the glomerulus can be viewed with the 3-4 serial sections lying randomly across the grid bars. We do not need a picture of the whole glomerulus, rather most pictures are between 3,000 and 10,000X. Dr. Tibor Nadasdy is the renal pathologist and decided last year that all our renal biopsies would be captured with the digital camera onto a computer and sent up to him via a network to his computer. So, at the present time we use very little EM film. He diagnoses each biopsy and e-mails representative digitized images to the nephrologists.
Karen L. Jensen, M.S. Project Manager & Associate Scientist Electron Microscopy Research Core
-----Original Message----- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, March 30, 2001 2:20 PM To: microscopy-at-sparc5.microscopy.com
Hello -
Has anyone out there tried to adhere xylem sections to pre-coated microscope slides like Fisher Probe-On Plus slides? I've tried and had a zero percent section retention. I can imagine that this is because of the scarcity of live cells (and plasma membranes). I've tried slow air drying and various temperatures on the slide warmer. The sections are 15 microns, and are from fixed (buffered paraformaldehyde) but not embedded samples. I'm about to try Poly-L-lysine and amino-acyl silane treated slides, but I'm not too hopeful because they (at least Poly-L) rely on the same positively-charged surface principle. I want to avoid gelatin or albumin subbing because I'm treating sections with protease, and also want to minimize background staining. Any tips would be greatly appreciated.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Rishi Raj wrote: ============================================================ We have just acquired a used microscope made by ISI Inc. (their model alpha - 1980). Would anyone please have information on how we can obtain spare parts, filaments etc., for this machine. Many thanks for your help... ============================================================ The business of the ISI SEM's is now being handled by
Aspex Instruments LLC Formerly: RJ Lee Instruments Ltd. 175 Sheffield Drive Delmont, PA 15626 USA Tel: 1-724-468-5400 Fax: 1-724-468-0225 E-mail: pssales-at-rjleeinst.com
The former manager of the SEM operation when it was still ISI, Michael McCarthy, is now with Aspex. Michael might be the single most knowledgeable person in terms of spare parts for the column and vacuum system. Several of the main suppliers of consumables, like SPI Supplies, also offer filaments, new and retipped, apertures, and the other items of that nature you would be needing to maintain the microscope.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I desperately want to be a materials researcher/Electron Microscopist. We have a Newly established Electron Microscope Laboratory. We need knowledge and skills. Please assist for a scholarship/suport as we have no funds, nor courses of the like at our University.
Hello everybody, We have an JEOL JXA 8600 EPMA in our Institute, with three WDS spectrometers. We are planning to buy an other one, and we where thinking about the H-type x-ray spectrometer.
Is there anybody in the list who has experience with this kind of spectrometers that could give me some information about them (their performance in general and also comparatively to standard spectrometers, etc.)?
Thanks
Laura Hernandez Laura Hernandez Laboratorio Microsonda Electronica Instituto GEA Universidad de Concepcion Casilla 160C Concepcion CHILE
Try using Vectabond treated slides. vectabond is available from vector laboratories. We had quite good results with leaf, stem, root sections sticking to these slides. Vector lab: 1-800-227-6666
Good luck,
Soumitra
I have no financial interest in Vector Laboratories.
On Sat, 31 Mar 2001, Rachel Spicer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello - } } Has anyone out there tried to adhere xylem sections to pre-coated } microscope slides like Fisher Probe-On Plus slides? I've tried and had a } zero percent section retention. I can imagine that this is because of the } scarcity of live cells (and plasma membranes). I've tried slow air drying } and various temperatures on the slide warmer. The sections are 15 microns, } and are from fixed (buffered paraformaldehyde) but not embedded samples. } I'm about to try Poly-L-lysine and amino-acyl silane treated slides, but } I'm not too hopeful because they (at least Poly-L) rely on the same } positively-charged surface principle. I want to avoid gelatin or albumin } subbing because I'm treating sections with protease, and also want to } minimize background staining. Any tips would be greatly appreciated. } } Rachel } } } } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
You can do a brightness leveling image process with a number of a standard packages.
The procedure is simply to do a Gaussian blur that gives you the slowly varying component of the background and subtract that from your original image.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Feng Wu [mailto:fwu-at-bgumail.bgu.ac.il] } Sent: Wednesday, March 28, 2001 6:14 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: adjust the intensity of the image } Sensitivity: Confidential } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hi, All, } I take some diffraction contrast pctures. As the thickness of } the samples } changes sharply the brightness of the images is not uniform. } Does someone know } any software to adjust the brightness for the whole image? } Thanks inadvance. } Best regards. } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } } fwu-at-bgumail.bgu.ac.il } ********************************************** } } }
I'm posting this message for a colleague; as always, I appreciate your help. Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL Richland, WA (509) 372-0692
} My graduate student has made TEM images of our plasma polymerized aniline } films. The films seem to have a "cauliflower" structure that could } probably be described as a fractal pattern. Have you ever made plasma } polymerized aniline films and if so did you see a cauliflower structure? } Your comments would be helpful. Thanks. } Pat } } Patrick D. Pedrow, pedrow-at-eecs.wsu.edu, www.eecs.wsu.edu/~pedrow
} Greetings, } I have found that polyethlyene-imine (PEI) is much stickier } than poly-lysine. I have no idea about the resin you mentioned, but } PEI is sticky stuff. It comes as a liquid. I make a 0.1% soloution in } ddwater, which I freeze in aliquots. Then I keep a working one in the } fridge. I coat coverslips in the stuff by floating them on a drop of } the PEI solution for about 10 sec, and then blotting off the excess } and letting them air dry. In that conditions, the coated 'slips are } good for at least months. } } Hope this helps, } Tobias Baskin }
Tobias:
Would this be Sigma catalogue number P-3143?
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I am looking for a X-ray radiation detector used on TEMs. Currently we have a Geiger counter that is only sensitive to low energy X-ray. The purpose of the new detector is to sense high energy X-ray leakage from a 200 kV scope.
Any suggestion or clue will be greatly appreciated. Please contact off-line.
Dear Haifeng, The x-ray monitors on our HVEM are wrapped in a metalic sheath so they will be sensitive to the brehmsstrahlung spectrum of 1.2 MeV electrons. I don't think that they could be easily connected to your scope, but you might look into making a similar sheath for your existing Geiger counter. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I have worked with CAMECA SX50 with the horizontal spectrometer. It was helpful when working with fractures, for example for identification of nonmetallic inclusions. Another it's advantage was that it was the only spectrometer with all 4 crystals, so it had more flexibility in maps or line scans acquisition. In quantitative analysis I did not use it both major and trace elements acquisition and never got any problems with its performance.
Vladimir Dusevich
-----Original Message----- } From: Laura Hernandez To: Microscopy-at-sparc5.microscopy.com Sent: 4/2/01 7:41 AM
Hello everybody, We have an JEOL JXA 8600 EPMA in our Institute, with three WDS spectrometers. We are planning to buy an other one, and we where thinking about the H-type x-ray spectrometer.
Is there anybody in the list who has experience with this kind of spectrometers that could give me some information about them (their performance in general and also comparatively to standard spectrometers, etc.)?
Thanks
Laura Hernandez Laura Hernandez Laboratorio Microsonda Electronica Instituto GEA Universidad de Concepcion Casilla 160C Concepcion CHILE
I am trying to determine a method to create or embed points of reference on an SEM sample. To give some background, the samples are sections of microprocessor packages that are placed in a modified stage with a three-point bend fixture. What I'm trying to do is monitor the movement of points on the specimen surface as the load is increased. I tried using the spot mode on the scope to see if I could remove some of the sputter coat, since most of the material underneath is non-conductive, but that was unsuccessful.
Imaging will most likely be done between 300 and 1000x. I need to have multiple reference points on the screen at one time, since I will be measuring relative displacements. The reference points do not need to be distributed in a uniform pattern.
I have tried applying some powder to the surface, since I read about this being done before, but the results were not acceptable. I may just be using the wrong type of powder, but I haven't been able to find out what kind of powders would work best
So, any ideas on how I may accomplish this?
Thanks,
Norman Kay Graduate Student AME Dept. The University of Arizona
I want to put a (cheap) video camera onto the optical microscope of my JEOL 840.
I want to leave the eyepiece lens on, so that users can have the option of easily removing the camera to use the microscope conventionally.
I have tried presenting several different models of CCD video cameras up to the eyepiece, both with and without the camera lens attached, but none gives me anything better than a smallish bright circle in the centre of the (black) field of view.
I presume that I need some sort of intermediate lens, but my understanding of physical optics has largely evaporated over the years.
Can someone point me towards a suitable text or other information source?
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } I want to put a (cheap) video camera onto the optical microscope of } my JEOL 840. } } I want to leave the eyepiece lens on, so that users can have the } option of easily removing the camera to use the microscope } conventionally. } } I have tried presenting several different models of CCD video cameras } up to the eyepiece, both with and without the camera lens attached, } but none gives me anything better than a smallish bright circle in } the centre of the (black) field of view. } } I presume that I need some sort of intermediate lens, but my } understanding of physical optics has largely evaporated over the } years. } } Can someone point me towards a suitable text or other information } source? } } thanks } } rtch
A CCD camera without a lens looking into and eyepiece usually has the opposite problem of having too much magnification. The image coverage of the image on the CCD camera can be increased when no lens is present on the camera simply by moving the camera further away from the eyepiece. My set up uses a 2.6x eyepiece for the video camera and a 10 x eyepiece to view the distance between the 2.6 eyepiece and the CCD element is about 2 inches or a little more and I have almost twice the magnification on the CCD camera as I see through the 10X eyepiece.
So just adding a spacer between your camera and your eyepiece should solve your problem.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
If you have coat your specimen with a thin carbon layer for conduction then add another layer of gold through a TEM grid as a mask you should be able to see the grid bars. Check out your local EM supplier's catalogue for the most suitable grid design.
Good luck, Ron
On Mon, 2 Apr 2001 15:48:48 -0700 SEM Machine {SEM-at-ACATC.AME.Arizona.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying to determine a method to create or embed points of reference on } an SEM sample. To give some background, the samples are sections of } microprocessor packages that are placed in a modified stage with a } three-point bend fixture. What I'm trying to do is monitor the movement of } points on the specimen surface as the load is increased. I tried using the } spot mode on the scope to see if I could remove some of the sputter coat, } since most of the material underneath is non-conductive, but that was } unsuccessful. } } Imaging will most likely be done between 300 and 1000x. I need to have } multiple reference points on the screen at one time, since I will be } measuring relative displacements. The reference points do not need to be } distributed in a uniform pattern. } } I have tried applying some powder to the surface, since I read about this } being done before, but the results were not acceptable. I may just be using } the wrong type of powder, but I haven't been able to find out what kind of } powders would work best } } So, any ideas on how I may accomplish this? } } } Thanks, } } Norman Kay } Graduate Student } AME Dept. } The University of Arizona } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
The magnifications you are using present the problem. I'm sure that you are also wanting as much resolution as possible from image processing of the resultant images.
Producing a non-conductive spot on the sample is a good idea, as it should stand out well, particularly during a slow record mode scan.
My vote - copier or laser printer toner. Very small particle size, non-conductive plastic composition. Also, once the powder is sprinkled on, it can be adhered to the surface with a little heat to ensure that it doesn't move around.
On Monday, April 02, 2001 5:49 PM, SEM Machine [SMTP:SEM-at-ACATC.AME.Arizona.edu] wrote: } } } I am trying to determine a method to create or embed points of reference on } an SEM sample. To give some background, the samples are sections of } microprocessor packages that are placed in a modified stage with a } three-point bend fixture. What I'm trying to do is monitor the movement of } points on the specimen surface as the load is increased. I tried using the } spot mode on the scope to see if I could remove some of the sputter coat, } since most of the material underneath is non-conductive, but that was } unsuccessful. } } Imaging will most likely be done between 300 and 1000x. I need to have } multiple reference points on the screen at one time, since I will be } measuring relative displacements. The reference points do not need to be } distributed in a uniform pattern. } } I have tried applying some powder to the surface, since I read about this } being done before, but the results were not acceptable. I may just be using } the wrong type of powder, but I haven't been able to find out what kind of } powders would work best } } So, any ideas on how I may accomplish this? } } } Thanks, } } Norman Kay } Graduate Student } AME Dept. } The University of Arizona } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
My collegue C. Ulhaq want to know who practice Lorentz Microscopy on TEM (in Europe particulary).
The questions would be : which kind of microscope you use, do you use special polar pieces, and did you buy it or were they "home made". Same question about the sample holder. What are the max magnification accessible ? We have a ABT Topcon 002B, with the the possibility to change the polar pieces.
You can answer direct to my collegue (corinne.ulhaq-at-ipcms.u-strasbg.fr), or on the list.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Thanks for the replies and the advice that you have offered.
I have Some additional information. I am a BSc.[Physics, Mathematics] holder. Currently I am working at the E.M laboratory of the university of Dar es Salaam as a supporting staff, and I am looking for the opportunity to pursue an MSc. and hence a Ph.D that will qualify me to research fellow. Funds are a hindrance. We have one old ZEISS9S2 TEM, and a new LEO 910 analytical TEM. An MSc. course in TEM or a research in metals/ceramics will do.
Hello, We have a client who needs to embed a cell culture in paraffin. In our lab we embed cultures in agarose and then embed for TEM. We do not have experience embedding paraffin. My question is, since xylene is used for paraffin how do you keep the culture from dispersing. Any suggestions? Thank you
Karen Kelley Senior Electron Microscopist University of Florida ICBR Electron Microscopy Core Lab Box 118525 Gainesville Florida Lab: 352-392-1184 fax: 352-846-0251 email: klk-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/staff/karenpage.html
I have not done Lorentz microscopy (here LEM) in a long time and I am not very familiar with the configuration of the Topcon 002B. This is what I remember ,for LEM to work the sample has to be outside the strong magnetic field of the OL pole piece. In the good old days, we used a modified top entry-type specimen holder which was longer than usual so that the specimen would sit below its normal position. In addition, the IL electronics had to be modified so that focus could still be attained when the specimen was in this lower position. We normally used magnifications of up to about 20 KX if I recall.
The second way of doing this is to essentially turn off the objective lens , but you have to be able to focus with the IL . Some of the newer scopes might not be able to do this without modifications to the electronics. In our case all the modifications were done by the manufacturer.
Hope this helps
Jordi Marti
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Tuesday, April 03, 2001 3:25 AM To: Microscopy Society of America
Hi everyone, We are working with suspension cells (Jurkats or H9s) and we need a protocol to prepare them for fluorescent microscope (both fixed and live cell imaging). Thanks in advance for your help.
Asli Oztan
asost2-at-pitt.edu University of Pittsburgh Molecular Virology and Microbiology
You can try spinning (centrifuging) the cells down so they become a packed ball of cells, then carefully put them into a bag (i think they are nylon bags) for processing and subsequently into paraffin block.
Frank Lee
Karen Kelley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } We have a client who needs to embed a cell culture in paraffin. In our lab } we embed cultures in agarose and then embed for TEM. We do not have } experience embedding paraffin. My question is, since xylene is used for } paraffin how do you keep the culture from dispersing. Any suggestions? } Thank you } } Karen Kelley } Senior Electron Microscopist } University of Florida } ICBR Electron Microscopy Core Lab } Box 118525 Gainesville Florida } Lab: 352-392-1184 fax: 352-846-0251 } email: klk-at-biotech.ufl.edu } http://www.biotech.ufl.edu/~emcl/staff/karenpage.html
Dear Norman, When we had a similar problem of creating a reference for studying crack growth, we used the gold-evaporated grid method, as Ron Doole mentioned. However, this gave problems because the grid is uniform, so after you've traveled a little way, you had no unique reference to keep you placed. We solved this by also puting a drop of latex sphere suspension on the surface, before sputter coating. This is a suspension of latex spheres of specific size, I believe we used one micron, but you can use a size suitable to your magnification. The suspension dries to form a random pattern of dots that can be compared in photos. You amy have to experiment with the concentration of spheres to get the right coverage. We were lucky enough to have a researcher making latex spheres who gave us some of her duds, but these suspensions can be purchased. I hope this helps. At 03:48 PM 4/2/01 -0700, you wrote: } } I am trying to determine a method to create or embed points of reference on } an SEM sample. To give some background, the samples are sections of } microprocessor packages that are placed in a modified stage with a } three-point bend fixture. What I'm trying to do is monitor the movement of } points on the specimen surface as the load is increased. I tried using the } spot mode on the scope to see if I could remove some of the sputter coat, } since most of the material underneath is non-conductive, but that was } unsuccessful. } } Imaging will most likely be done between 300 and 1000x. I need to have } multiple reference points on the screen at one time, since I will be } measuring relative displacements. The reference points do not need to be } distributed in a uniform pattern. } } I have tried applying some powder to the surface, since I read about this } being done before, but the results were not acceptable. I may just be using } the wrong type of powder, but I haven't been able to find out what kind of } powders would work best } } So, any ideas on how I may accomplish this? } } } Thanks, } } Norman Kay } Graduate Student } AME Dept. } The University of Arizona } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Can someone identify a supplier for small amounts of methlyamine tungstate powder (used for negative staining in TEM)? I have tried EMS, Ted Pella, and Polysciences (the supplier of our original decade-old vial). Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
I'm trying to get a handle on the value of an old Auger/ESCA spectrometer. The model is PHI 558, by Perkin-Elmer. It was built around 1985 or 86, and the electronics are quite old, but it's intact and fully functional. It also has a LEED detector and has been upgraded over time. A partial list of parts is included below. Does anyone have an idea what this might be worth?
Thank you, Ellen Carrillo-Heian
emheian-at-engr.ucdavis.edu Dept. of Chem. Eng. and Mat. Sci. UC Davis Davis, CA 95616 USA ---------------------- Partial list of components:
} 32-010 Lock In Amp } 20-0275 Electron Multiplier Supply } 32-095 X Ray Source Control } 22-040 DC Power Supply } 16-020 Heat Exchange / Deionizer } 20-805 Analyzer Control } 32-100 Electron Multiplier Supply } 11-065 Ion Gun Control } 20-115 Ion Gun Control } 11-010 Electron Gun Control } 11-055 ESCA / Auger System Control } 11-500A Auger System Control } Inficon 012-214 } 04-303 Differential Ion Gun } 04-548 Dual Anode X-ray Source } 15-255 GAR Precision Electron Energy Analyzer } Ultek DI Pump } 218075B-26 UHV Instruments (Tag in front of the screw/bellows assembly, on } frame.) } } And a 386 Compaq controlling it. } } The chamber has the dual chamber translation set up. (With the 4 ft } screw/bellows) }
We do this using contamination spots from the microscope itself. We put down a grid array of spots using our Isis system to control the stage and spot positons. Focus the beam to a spot and let is sit there for a minute or so and a contamination spot should appear. We are using electropolished aluminum. I dont know if it will work for your material but its worth a try Then we strain our material and look at it again. Works well.
I would like to do high resolution TEM on spherical, semiconductor nanoparticles. The diameters of these particles range from 1 to 10 nm. Our goal is to acquire good digital images of these particles so that we can size them in house. Please let me know if you can help and how much you charge per sample or per hour. Your help will be greatly appreciated.
Dear Dr. Palmer, Nanoprobes sells the methylamine tungstate you desired under the name "NanoW". It is an excellent negative stain. They also make methylamine vanadate, a similar, but lower atomic number stain they call "NanoVan". More information is at www.nanoprobes.com. J. Hainfeld
Dr. James F. Hainfeld Brookhaven National Laboratory Biology Dept. Bldg. 463 Upton, NY 11973 USA Tel. 631-344-3372 Fax. 631-344-3407 email: hainfeld-at-bnl.gov website: http://bnlstb.bio.bnl.gov/biodocs/stem/stem.html
Generally, equipment depreciates at the rate of 30% per year on the balance. In other words, an SEM with an initial cost of 100K is worth 70K after the first year, 49K the second year, 34.3k the third year, etc.
These numbers are based upon the experience I have had with used equipment sales during the past five years. Other allowances are made for equipment that has been abused, etc. Strangely enough, accessories add little to the resale value of equipment.
I am sure there are others who would differ as it would not fit into their accounting procedures but my experience has been that scientific equipment depreciates more than computers.
Regards,
Earl Weltmer
I have no financial interest in this thread only experience.
----- Original Message ----- } From: "Ellen Carrillo-Heian" {emheian-at-engr.ucdavis.edu} To: "microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 03, 2001 11:25 AM
Hi everybody (again),
I am looking for a program that allows me to quantify a given group of nanoparticles that have various shapes and sizes. These particles range from 1 to 10 nm in diameter. Your help will be greatly appreciated. Thank you.
We sell this as a 2 % aqueous solution (suitable for use directly) - the product is called "Nano-W." It's listed on our web site catalog under "Negative staining."
Regards,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
What kind of computer depreciates (looses value) slower than scientific equipment? A regular PC or Mac drops by over 50% the first year. After an additional six months, the system worth next to nothing. But of course, the new and improved model is $2K or more. Either way, the standard IRS depreciation schedule for computers and scientific equipment is five years. I would say that this is more appropriate for scientific equipment than it is for computers. But YMMV.
gary g.
At 05:16 PM 4/3/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is an interesting thread. Our unit is supposed to operate at "full cost recovery", and as such we are charged depreciation of the instruments (funded from a central equipment grant), which are depreciated over 15 years. Only computers are depreciated over 3 years. If anyone else is aware of some vaguely standard depreciation times for TEMs, SEMs, confocal, light microscopes, I'd appreciate hearing about this.
Thanks, rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
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Rosemary said - } This is an interesting thread. Our unit is supposed to operate at "full } cost recovery", and as such we are charged depreciation of the instruments } (funded from a central equipment grant), which are depreciated over 15 } years.
FWIW, I understand that that our organization (A Canadian federal government one) also depreciates this kind of major capital equipment over 15 years. (Except, apparently, helicopters - our Navy is still using ones which are, literally, older than most of the pilots flying them....)
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
John Russ recently demonstrated an adaptive equalization technique for us that works quite well. Basically it is histogram equalization over a local area of the image that is then stepped over the whole image. He has implemented this method in his IP Toolkit and Fovea Pro packages that plugin to PhotoShop. It is also described in The Image Processing Handbook, 2nd edition on page 222.
Just a satisfied customer...
Henk Colijn
At 11:42 AM 4/2/01 -0400, Walck, Scott D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
An interesting thread indeed! We are in the midst of surplusing out a lot of used histology equipment, and find that there are two different methods of determining value. Depreciation of equipment for tax purposes is done according to state/federal tax laws--and I think that means after 5 or 10 years (the time seems to change, and I can never keep up with) the _tax_ value is zero. However. When it comes to disposing of the equipment, our business folks are using a different "book value" that has values significantly greater than current market value. So.... Draw your own conclusions.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Wed, 4 Apr 2001 15:47:39 +1000, Rosemary White wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | This is an interesting thread. Our unit is supposed to operate at "full | cost recovery", and as such we are charged depreciation of the instruments | (funded from a central equipment grant), which are depreciated over 15 | years. Only computers are depreciated over 3 years. If anyone else is | aware of some vaguely standard depreciation times for TEMs, SEMs, confocal, | light microscopes, I'd appreciate hearing about this. | | Thanks, | rosemary | | | Rosemary White | Microscopy Centre | CSIRO Plant Industry | GPO Box 1600 | Canberra, ACT 2601 | Australia | | phone 61-2-6246 5475 | fax 61-2-6246 5000 | email r.white-at-pi.csiro.au | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I think that CSIRO is on the high side. In the Canadian government 10 years seems to have been the semi-official standard for as long as I can remember. In my lab, though, we informally think of two 'effective lifetimes', 7 years or thereabouts for the TEM and SEM, and 10 for the other beam instruments (EPMA, SIMS, XPS and SAM). Furthermore, we regard these as upper limits for two reasons:
- vendors these days are operating on ever-tighter parts inventories, thus a beam instrument may have some good years left in it, but you find you can't get a crucial part any longer. This happened to us recently with our 12 year old Cameca SIMS, when a flight tube developed ultrafine cracks and we discovered that they are not kept in stock any more. After a lot of arm-twisting and calling of favors, we tracked down one of the few remaining ones in Europe, otherwise we would have been down for 3 months awaiting a custom-built one.
- we do a lot of work with the private sector, some on a contract basis. They often come to us to get state-of-the-art data quality which has been collected in a timely fashion. The first is what usually gets their attention in the first place, while the second is crucial to keeping their attention.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Rosemary White } Sent: Wednesday, April 04, 2001 1:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: equipment depreciation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is an interesting thread. Our unit is supposed to operate at "full } cost recovery", and as such we are charged depreciation of the instruments } (funded from a central equipment grant), which are depreciated over 15 } years. Only computers are depreciated over 3 years. If anyone else is } aware of some vaguely standard depreciation times for TEMs, SEMs, } confocal, } light microscopes, I'd appreciate hearing about this. } } Thanks, } rosemary } } } Rosemary White } Microscopy Centre } CSIRO Plant Industry } GPO Box 1600 } Canberra, ACT 2601 } Australia } } phone 61-2-6246 5475 } fax 61-2-6246 5000 } email r.white-at-pi.csiro.au } } }
I am processing samples of white fat for TEM and have been having trouble with a dense precipitate that covers the cytoplasm. Nuclei and blood vessels are relatively unaffected. So far I've tried 3 different fixatives: Trump's and 2.5% glut/2% para in either 0.1M cacodylate of 0.1M phosphate buffer. The samples have been osmicated for one hour and embedded in either Spurr's resin or Epon. The precipitate is present in unstained sections as well as those stained with UA alone or UA and lead. Treatment with .5N HCl for 2 minutes alleviates the problem somewhat but bleaches the sections so much that they are very difficult to see. If anyone has encountered similar problems and has suggestions for me I would be most grateful.
Thanks in advance
Germaine G. Boucher TEM Lab Pfizer Global Research and Development Groton, CT
We are looking to replace a very old EDS system on our Hitachi S-2460N variable pressure SEM. I would like to know about your preferences for and experiences with different companies, both good and bad. Please reply directly to me. If anyone else is interested, I could submit a summary to the list after I've collected all the responses.
Thanks in advance.
Jean Ross Central Microscopy Research Facility University of Iowa
I have two questions concerning service contracts on confocals. Let me start by saying i have had a confocal for about 8 years and would never consider going without one. I have a Biorad 2000 going off warranty and need to make a decision.
First question: Biorad no longer guarantees a response time - they now promise to get to you as fast as they can but no longer promise a 48 or 72 hr response. Have other confocal manufacturers done this also?
Second question: My university is pushing replacing service contracts with "insurance" contracts with a major vendor who then pays for a service visit from the manufacturer on an hourly basis. All parts, travel, service repair time, etc are covered at a price that is typically 75% less than the manufacturer's service contract. They guarantee the price and coverage for 3 years. Personally I don't know how they could make money on this deal since we average a fair number of visits and spare parts (e.g. lasers) in a typical year. Does anyone have experience with this type of situation with confocals? The company the University is dealing with is CIC but there are several other ones out there.
Thanks for any input. -- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
dear listservers.... i need some advice.... we use Permount as our mountant...however, as we cut our slides extremely thick (120 micra) for our staining method (Golgi-impregnations of neurons), it often takes 2-3 weeks for the Permount to dry sufficiently so that we can actually use the slides without it getting on our microscope stage, or the coverslip moving around under our oil-immersion lenses.... so....my question is --- does any microscope maven out there know if there is anything that can be done to accelerate the drying/hardening the Permount....??? many thanks for any help.... regards, Ron Mervis ~~~~~~~~~~ Ronald F. Mervis, Ph.D. Neuro-Cognitive Research Laboratories 2109 West Fifth Avenue Columbus, Ohio 43212 USA ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ..independent nonprofit contract laboratories dedicated to quantitiative neurostructural analysis to promote our knowledge and understanding of human neurological diseases, neurodegeneration, and neuroplasticity.... ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Tel: (614)-486-6404; lab: (614)-486-6080 Fax: (614)-486-6020 e-mail: RonMervis-at-aol.com (or) RonMervis-at-Neuro-Cognitive.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ "...can the human soul be glimpsed through a microscope? Maybe, but you'd definitely need one of those very good ones with two eyepieces." - Woody Allen
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} dear listservers.... {BR} i need some advice.... {BR} we use Permount as our mountant...however, as we cut our slides extremely {BR} thick (120 micra) for our staining method (Golgi-impregnations of neurons), {BR} it often takes 2-3 weeks for the Permount to dry sufficiently so that we can {BR} actually use the slides without it getting on our microscope stage, or the {BR} coverslip moving around under our oil-immersion lenses.... {BR} so....my question is --- does any microscope maven out there know if there is {BR} anything that can be done to accelerate the drying/hardening the {BR} Permount....??? {BR} many thanks for any help.... {BR} regards, {BR} Ron Mervis {BR} ~~~~~~~~~~ {BR} Ronald F. Mervis, Ph.D. {BR} Neuro-Cognitive Research Laboratories {BR} 2109 West Fifth Avenue {BR} Columbus, Ohio 43212 USA {BR} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ {BR} ...independent nonprofit contract laboratories dedicated to quantitiative {BR} neurostructural analysis to promote our knowledge and understanding of human {BR} neurological diseases, neurodegeneration, and neuroplasticity.... {BR} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ {BR} Tel: (614)-486-6404; lab: (614)-486-6080 {BR} Fax: (614)-486-6020 {BR} e-mail: RonMervis-at-aol.com (or) RonMervis-at-Neuro-Cognitive.org {BR} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ {BR} "...can the human soul be glimpsed through a microscope? Maybe, but you'd {BR} definitely need one of those very good ones with two eyepieces." {BR} - Woody Allen {BR} {/FONT} {/HTML}
I have seen some beautiful SEM pictures of semiconductor interconnect lines (Copper and aluminum) in which the dielectric material (Silicon dioxide I assume) has been completely removed. Can someone tell me what was used to remove the dielectric without affecting the metallization?
Hello friends, I have a gold bronze composition material coming in for imaging by SEM, x-ray mapping, and optical microscopy. I could us a sample prep recommendation, particularly for a etchant or so I think. This is primarily a failure analysis project in which we want to clearly observe and differentiate grain boundaries. Recommendations would be greatly appreciated.
thanks, Bruce Brinson Optical Analyst Rice University
Germaine, If you don't rinse in buffer well enough after the initial fixation , the glut will form a precipitate with osmium. Try washing 4x or 5x for 15 minutes each between glutaraldehyde and osmium. Good luck,
At 10:34 AM 4/4/01 -0400, Boucher, Germaine G wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
Thanks for all the recommendations for suppliers of cameras and interfaces, but what I was wanting was a pointer to a text or somesuch from which I could figure out myself what I need.
There must be someone in this learned and experienced community who's been there and done that, isn't there?
"That", for those who may have missed my first post, being the problem of how to work out what sort of intermediate lens would be needed to interface a small cheap CCD video camera (or a webcam) so that it gives a good image when looking into the existing eyepiece lens of a given optical microscope (in this case the OM of a JEOL 840 SEM).
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear colleagues, I would like to buy a digital camera for my Axiolab Zeiss microscope. Unfortunately I am a bit confused in the amount of available data. I would need a digital camera of the resolution that matches the quality of film cameras in order I need not scan photos or negatives. Could you please be so kind and give me a piece of advice? I have Olympus Camedia 3030 with 3,34 mil pixels. Will this camera and resolution do or do I need to buy another one? If yes of what resolution and type? I will be very indebted for an advice because I receive often contradicting information for different distributors and I am not much wise about it. Many thanks in advance. With best wishes Jiri Kalvoda Department of Geology and Paleontology Masaryk University Kotlarska 2 61137 Brno Czech republic
It is easily done with a plasma etch using CF4 + O2.
The top layer of "glass" is typically silicon nitride over silicon dioxide or in earlier devices, it is boron phosphor silicon glass (BPSG).
I have some colorized shots on my web site at:
http://photoweb.net
More get added and some get swapped over time.
gary g.
At 11:48 AM 4/4/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rod McCabe wrote: ======================================================= I have seen some beautiful SEM pictures of semiconductor interconnect lines (Copper and aluminum) in which the dielectric material (Silicon dioxide I assume) has been completely removed. Can someone tell me what was used to remove the dielectric without affecting the metallization? ======================================================= While HF and Q-Tips can be used to swab off the SiO2, the better way (in our opinion) is with reactive plasma etching, using CF4 as the reactive gas. This way the layer is removed in a way that does not disrurb, for example, corrosion product that might have formed underneath. If you use the wet chemical approach, you can dissolve and swab away features of interest, such as corrosion product. And you have removed that which you might otherwise have been able to analyze with EDS or even Auger.
Several manufacturers offer table top plasma etchers, SPI Supplies being one of them. You can find information about the SPI Plasma Prep™ II on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am writing for comments -- off-list please -- about the CleanZone LF clean air workstation manufactured by IQAir, which, I am told, is used in Switzerland and Germany, but only recently has been introduced in the United States. With thanks,
James Martin Orion Analytical, LLC www.orionanalytical.com martin-at-orionanalytical.com
A colleague has asked for recommendations for setting up a digital darkroom (fun to spend someone else's money!). This person would benefit from a really good scanner that could deal with prints, large format negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked into an Agfa Duoscan T2500. Do any of you have an opinion about this or other suitable scanners?
I know this subject comes up regularly, but I don't feel bad about introducing it again, since technology evolves so quickly!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
thanks to everyone for the advices about all type of glues for tripod polishing. I tried to make some sort of summary from the different answers I have received. I would be happy if it could be any use for other tripod beginners as well.
Cheers! Csaba
1.) The best mail which decribes well the everyday life and joy of a microscopist comes first.
You really can't look at a spec sheet and know that it works - it's all trial and error.
2.) About the type of glue to be used:
The best material to use is super glue which is a cyanoacrylate material. There are different types available under various names. They all have slightly different properties and vary in such things as viscosity. The most important point is that the super glue is fresh. Once it is opened, you can use it for a short time, but then need to replace it. Sometimes, the glue from an unopened package can also be bad if it has been on the shelf for a long time. The trouble with using consumer products is that they sometime change without warning.
You should buy it from a place that has a high turnover rate of it so that it is fresh. The IBM guys recommend keeping the stuff unopened in the refrigerator for any length of time. Once it has been opened, you can't use it for very long (a day or two). I have used the Loctite a little longer because it has a very good sealing top. The Loctite product is also available in a "pen" type applicator which seems to have the best bench life of any of the applicators I've used. My tube usually is swiped off the bench long before it goes bad.One of the advantages to the cyanoacrylic cement is it dissolves, in a reasonable time, in acetone.If you were to use a crosslinked epoxy, you'll need to devise a cleaning scheme that will exhaustively remove the epoxy without altering your sample.
Basically what we're looking for is an intermediate viscosity glue (somewhere between maple syrup and water) that bonds in about 30-40 seconds and is very strong. Don't use any type of super glue gel! It doesn't work, you have to use the thin stuff.
In the end we are using a Loctite Prism 460.
Crystalbond 509. This is an acetone soluble glue which melts at 150*C and becomes quite viscous. We use it quite often here at Queens as a temporary glue for ceramics that have to be ground and polished on all four sides. If the bond breaks you simply heat it up and reset the part. This stuff comes in sticks that last for a very long time. Their website is as follows. {http://www.aremco.com/}
I like to glue specimens with epoxy resin because the substance does not harden too fast, and you always have time to find the best location for your sample, so that the latter does not break.
I have tried the "super glue" approach with no luck. Many technicians advocate Lock Tite brand of super glue that many companies sell with the tripod polish kit. I have always utilized Crystalbond adhesive. It is a low melting point (77 C) wax that is dissolved in acetone. If your sample is heat sensitive, super glue is the only other choice I know.
3.)Glueing advices:
--the biggest reason for sections falling off is the cleanliness of the pedastal. It must be cleaned with clean solvents that do not leave any trace of a contamination film on the glass. Check for cleanliness by holding the tool such that light reflects from the surface.
--the suface that you are glueing to should be as rough as possible to obtain a tooth or larger surface area for the glue to adhere to.
--any way you can improve the surface area would be great.
--the same above would be for the specimen
--pressure on the sample while it cures might be important, but the IBM guys just wick the extra stuff away from the sample and don't put a lot of pressure on the sample.
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
I've followed with interest the discussion on equipment depreciation and service contracts . I've noticed over the years in both metals research and now the semiconductor industry with increasing sophistication/specialisation of equipment running a lab to a given budget seems to mean accepting manufacturers service contracts with fewer 'independent' sources being able or willing to offer maintenance assistance .
My question arises from a recent 'confocal service contract' letter and I wonder does an insurance contract service alternative as offered by CIC exist in the UK ? and if so could anyone let me know where I could get more information ? .
Martyn Harris Device Engineer ESM Ltd , Cardiff Rd Newport , South Wales UK NP10 8YJ .
First, a disclaimer. I am a third party service provider for a variety of equipment, but not confocals. I have worked for a number of manufacturers in the past and been self-employed for around 20 years.
In regards to the first question - service departments in general are getting squirrellier. Contract prices have gone up greatly over the last decade or so while quality and parts stores have gone down. A generalization, granted, but one I'd be happy to back up.
As for the second question - don't get me started again. Given recent problems with the industry in general, I can only suggest that your university carefully study the question and check with several of their customers who have been with them for at least a couple of years insuring similar equipment. That goes for any such provider. I've been a vocal antagonist here of the concept, and from what I have heard from some customers, perhaps rightfully so. Not right, so far, in my feelings regarding the potential long term problems. Rather in the broad approach they have taken. Perhaps in trying to be a jack of all trades, they are a master of none.
To those of you who might be surprised by my abnormally low tone in this posting, please understand that we are getting to a point in time where these organizations are getting quite large in a very short period of time. As with any company or industry, they do have problems that they will not publicize. They also have increasingly large budgets for legal teams that would probably be anxious to root out any libel. They do save many organizations large amounts of money, but you have to ask, at what cost?
On Wednesday, April 04, 2001 10:20 AM, Tom Phillips [SMTP:PhillipsT-at-missouri.edu] wrote: } } } I have two questions concerning service contracts on confocals. Let } me start by saying i have had a confocal for about 8 years and would } never consider going without one. I have a Biorad 2000 going off } warranty and need to make a decision. } } First question: Biorad no longer guarantees a response time - they } now promise to get to you as fast as they can but no longer promise a } 48 or 72 hr response. Have other confocal manufacturers done this } also? } } Second question: My university is pushing replacing service } contracts with "insurance" contracts with a major vendor who then } pays for a service visit from the manufacturer on an hourly basis. } All parts, travel, service repair time, etc are covered at a price } that is typically 75% less than the manufacturer's service contract. } They guarantee the price and coverage for 3 years. Personally I } don't know how they could make money on this deal since we average a } fair number of visits and spare parts (e.g. lasers) in a typical } year. Does anyone have experience with this type of situation with } confocals? The company the University is dealing with is CIC but } there are several other ones out there. } } Thanks for any input. } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
I don't know if this is exactly what you are looking for, but Reetz and Coworkers have recently published a high troughput routine for nanaoparticle analysis: Reetz, Manfred T.; Maase, Matthias; Schilling, Tobias; Tesche, Bernd. Computer Image Processing of Transmission Electron Micrograph Pictures as a Fast and Reliable Tool To Analyze the Size of Nanoparticles. J. Phys. Chem. B (2000), 104(37), 8779-8781.
Good Luck,
Andreas
************************************************* Dr. Andreas Taubert Dept. of Materials Science and Engineering 3231 Walnut Street University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
Tina: We have the Duoscan 1200 but the 2500 is also a very nice unit--additional (real) resolution and a high O.D. range plus 14 or 16 bit image depth. The Agfa units also come with a built-in transparency plate (rather than having to add on a separate transparency adaptor). I find that color fidelity is very good with the Duoscan, and Agfa provides both reflective and transparency calibration standards. I am currently scanning in 3x4 TEM negatives at 12 bits (yield is about 26MB per image), and scan time is fairly rapid. There is one option that you might want to consider: the DIImage unit is made for up to 4x5 negatives, and I think (can't remember the last time I read the specs--the neurons aren't firing today) that resolution is in the 2700 dpi range--even for the 4x5 size.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
The opinions expressed are solely my own and do not constitute an endorsement of any vendor or manufacturer. I have no fiduciary interest in either company.
On Wed, 4 Apr 2001 17:22:23 -1000 (HST), Tina Carvalho wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi, All- | | A colleague has asked for recommendations for setting up a digital | darkroom (fun to spend someone else's money!). This person would benefit | from a really good scanner that could deal with prints, large format | negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked | into an Agfa Duoscan T2500. Do any of you have an opinion about this or | other suitable scanners? | | I know this subject comes up regularly, but I don't feel bad about | introducing it again, since technology evolves so quickly! | | Mahalo, | Tina | | http://www.pbrc.hawaii.edu/bemf/microangela | **************************************************************************** | * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * | * Biological Electron Microscope Facility * (808) 956-6251 * | * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* | **************************************************************************** | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce.
Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
Nominal grain size of film is about 10 microns (varies with film speed etc but this is the right order of magnitude). Thus to digitize the film to it's nominal limits your scanner should be able to digitize to better than this spatial dimension.
A simple back of the envelope calculation says a spatial resolution of 10 microns is 2540 - dpi..... and as we all know that must be the optical resolution of the scanner not the interpolated resolution. Scanners at this end are obviously more than you need to digitize photo's and get expensive quickly. Also when you see 2 numbers listed as the scanners resolution, believe only the first number, that is the CCD resolution.
Now add your bit depth. 12 bits is the minimium I would shoot for greyscale image, but if your attempting diffraction work the higher the better (i.e. 14 -16 bits+). For color work obviously multiple the bit depth by 3 one for each primary color (RGB). I've seen a number of 36 bit color scanners but not too many 48 bit ones at } 2540 dpi.
Lastly, dit depth is irrelevant if you don't have a high optical density capabilities otherwise your just digitizing noise. The highest value I believe is an OD of 4.0 but this is for DRUM scanners. Flatbed scanners typically run as low as 2.8, upwards to about 3.4 for the best I've seen in a flatbed.
At 09:40 AM 4/5/01 +0000, Ritchie Sims wrote: } "That", for those who may have missed my first post, being the } problem of how to work out what sort of intermediate lens would be } needed to interface a small cheap CCD video camera (or a webcam) so } that it gives a good image when looking into the existing eyepiece } lens of a given optical microscope (in this case the OM of a JEOL 840 } SEM).
You might try a web search for "afocal coupling". See http://www.photosolve.com/xtendascope.asp for example.
This page, and the message below, are for coupling to the eyepiece of telescopes, but the process should be similar with a microscope eyepiece. If you're looking for "small and cheap", put in a wide field eyepiece and hold the camera close to it and see what happens. I suspect you'll have some cropping of the image due to optical mismatch.
- John
} A friend of mind recently bought a fairly expensive SLR digital still } camera, an Olympus C-2500L, } } http://www.olympusamerica.com/product.asp?c=57&p=16&s=12&product=380 } } and has experimented with afocal photography through both his 8-inch f/6 } Dob and a microscope. All he sees in the camera is a small central disk } of light. He got the following explanation from a post on rec.photo. } digital. } } For cameras with LARGE taking lenses (f 2.8, f2.0) [...] there will } be a problem since their entry pupil ( the area of the lens that lets } the image into the lens) is so large compared to the exit pupil (the } opening on the eyepiece that lets the image out of the microscope) } that a large amount of the image will be lost!! This results in } severe vignetting, and so only a small central spot of image in a } dark field is recorded by the digital camera. } } Does this sound reasonable? The optics of this situation are pretty } mysterious to me, so I can't judge. But it seems like increasing the } focal length of the camera lens (zooming in) should compensate for the } effect of the small exit pupil, and that the problem might be more one } of eye relief (he just can't get the lens close enough to the eyepieces, } which I think are a couple of Plossls and the 25mm SMA that comes with } Celestron Dobs).
And here's the response so far.
} Subject: Re: afocal astrophoto problem: exit pupil? } Date: Wed, 4 Oct 2000 17:52:19 -0400 } From: "Michael A. Covington" {See http://www.CovingtonInnovations.com for address} } Organization: MindSpring Enterprises } Newsgroups: sci.astro.amateur } } } Does this sound reasonable? } } No. The camera lens is *supposed* to have a larger entrance pupil than the } exit pupil of the telescope. When doing afocal photography with an SLR the } entrance pupil of the lens is an inch or more in diameter. } } If there is vignetting, it's probably because the camera is the wrong } distance from the eyepiece -- either too close or too far. } } -- } } Clear skies, } } Michael A. Covington / AI Center / The University of Georgia } Author, ASTROPHOTOGRAPHY FOR THE AMATEUR } http://www.CovingtonInnovations.com/astro {} { } } } Subject: Re: afocal astrophoto problem: exit pupil? } Date: Wed, 4 Oct 2000 14:25:48 -0700 } From: "Bob May" {bobmay-at-nethere.com} } Organization: Posted via Supernews, http://www.supernews.com } Newsgroups: sci.astro.amateur } } Bet that when you look at the viewfinder, you will see an image just } like one that you would see if your eye were at a certain distance } from the EP. If that image looks like the eye is too far away from } the EP then it's a sure thing that the camera's lens is too far away } from the EP. That's how it's all done. The camera is nothing more } than an aritificial eye. } -- } Bob May } Remember that computers do exactly what you tell them to do, not what } you think that you told them! } Bob May } } } Subject: Re: afocal astrophoto problem: exit pupil? } Date: Wed, 04 Oct 2000 19:43:43 GMT } From: "Chuck Olson" {chuckolson01-at-home.com} } Organization: -at-Home Network } Newsgroups: sci.astro.amateur } } Yes, it is critical that the eyepiece and camera lens be somewhat } physically compatible with each other. The eyepiece must put the } exit pupil about in the plane of the camera iris opening, which } in most instances requires the eyepiece to be virtually in } contact with the camera lens front element. For instance, the } Nikon Coolpix 950 and 990 have relatively small lens fronts and a } nice 28mm (I think) thread that adapts readily to the T-thread } that is often used in astrophotography. As a result, the CP950 } easily looks through 17mm , 26mm, or 32mm Plossl eyepieces. Even } there, as you point out, the camera needs to be operating at the } tele end of its zoom range to fill the rectangular frame, rather } than showing a small, circular, fuzzy-edged, wide-angle field. } } I'm not sure what the C-2500L looks like, but it may have a } physically larger lens that has its iris deep behind the front } surface. This might require a very long focus eyepiece, like a } 40mm Plossl, conceivably, or one with even greater eye relief, to } accomplish the optical hook-up more favorably, and may limit the } operation of the overall system to somewhat lower magnifications. } Oh, once you have a compatible eyepiece, then you can use Barlow } lenses to get back needed magnification for your desired image } scale. The only probmen there is the setup gets pretty long as } these lenses are stacked up, and stability may suffer. } } The Nikon, as mentioned, has been found by many to be almost } ideal for afocal photography of the moon and planets. } } Chuck }
************************************************************************* {/b= igger} {/bold} Interest in the sophisticated fluorescence imaging techniques of Fluorescence Resonance Energy Transfer (FRET) and Fluorescent Lifetime Imaging Microscopy (FLIM) amongst the biological research community has grown in recent years. FRET imaging provides a tool to solve complex structural associations at resolution limits beyond conventional optical imaging. FLIM allows the measurement of FRET without the significant problems associated with intensity based FRET measurement, as well as faster, more accurate and quantitative measurement of cell physiology. These techniques are also being implemented in high-throughput screening regimes for drug discovery. Invited lectures by a distinguished group of scientists will concentrate on new technical developments in these areas and demonstrate successful application of these techniques in biological and industrial settings.=20
A poster session has been organized for June 9th so that registrants may present their experiences with FRET and FLIM. =20
We anticipate that this will be a most enjoyable as well as intellectually stimulating symposium.=20
************************************************************************* {/b= igger} {/bold} Interest in the sophisticated fluorescence imaging techniques of Fluorescence Resonance Energy Transfer (FRET) and Fluorescent Lifetime Imaging Microscopy (FLIM) amongst the biological research community has grown in recent years. FRET imaging provides a tool to solve complex structural associations at resolution limits beyond conventional optical imaging. FLIM allows the measurement of FRET without the significant problems associated with intensity based FRET measurement, as well as faster, more accurate and quantitative measurement of cell physiology. These techniques are also being implemented in high-throughput screening regimes for drug discovery. Invited lectures by a distinguished group of scientists will concentrate on new technical developments in these areas and demonstrate successful application of these techniques in biological and industrial settings.=20
A poster session has been organized for June 9th so that registrants may present their experiences with FRET and FLIM. =20
We anticipate that this will be a most enjoyable as well as intellectually stimulating symposium.=20
} Dear colleagues, I would like to buy a digital camera } for my Axiolab Zeiss microscope. } Unfortunately I am a bit confused in the amount of } available data. I would need a digital camera of the } resolution that matches the quality of film cameras } in order I need not scan photos or negatives. } ...
To give you an idea of what you are asking: For comparable resolution, the camera would need deliver more than 6M pixels ... and there is also the question of a digital camera capturing the gamut of color capable of film. For example, the camera you mention, which is aimed at consumers, probably delivers a gamut aimed at the "sRGB" color space. Only a film scanner can capture a color gamut comparable to "Ektaspace RGB". Still, your camera is likely to do a very good job if properly adapted to the microscope. You will need a 1X C-mount adapter for the microscope head, and an adapter for mounting the camera on the C-mount. These are readily obtained for Nikon Coolpix cameras, possibly yours too.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
Hello Tina, I have the scanner you are looking at & like it a lot. To be quite honest I do not find that I need to exploit it'll full capably. If I were in the market again, looking at newer technology I would be interested in a faster scanner of similar quality. Yes I want my cake & to eat it too :). I'll give you this analogy. If I have 10 negatives I will franchise my time, that is let things scan while I hang out in the office doing other things. If I have 20 negatives, I'll probably goto the darkroom to make photos. It is quicker & paper is cheaper. BTW I have an Epson 870 inkjet that produces nice quality images... cost is down to $180 US, (now the Epson 880)....no financial interest in these companies.
Oh yea, get the fastest computer you can afford.
good luck, Bruce Brinson Rice U.
Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, All- } } A colleague has asked for recommendations for setting up a digital } darkroom (fun to spend someone else's money!). This person would benefit } from a really good scanner that could deal with prints, large format } negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked } into an Agfa Duoscan T2500. Do any of you have an opinion about this or } other suitable scanners? } } I know this subject comes up regularly, but I don't feel bad about } introducing it again, since technology evolves so quickly! } } Mahalo, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
Tina, Have your colleague check out the Imacon Flextight Precision II scanner. The optical resolution is 5760 dpi for slide-sized objects; I believe it drops to 4800 dpi for objects the size of her larger negatives. The scanner collects 14 bits of usable data per channel, which can be exported as a two bytes per channel, and has a dynamic range of 3.9 OD units (4.1 OD max). The machine is also very fast. The URL is:
At 05:22 PM 4/4/2001 -1000, Tina Carvalho wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tina,
I have the Duoscan and a Nikon slide scanner. The Duoscan can scan slides on the special tray feature but side by side comparisons of the Duoscan and Nikon show that the Nikon scan is much better. For the larger negs we had a special tray made for the Duoscan and we scan in our EM negs. The Nikon has gotten much cheaper and an excellent scanner can be had for $700 with Digital ICE, something you want.
Get two scanners.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
I forgot to mention John that we do not use formar coated grids. For better assessment of the renal biopsy we do not mince the sample, either. Please see our Web page for more details. {pathology.lsuhsc.edu/Pathist/dx_home.htlm} click on M.diagnostic service and then on renal biopsy. Our diagnostic em lab continues to send poloroid HRLM pictures and TEM B&W contact prints with each report.
Lehigh University seeks an Electron Microscopy Technician to perform duties in support of the Microscopy Center of the Materials Science and Engineering Department. The person appointed will work with other technical staff to instruct students in the operation of microscopes and other equipment; maintain and repair instruments; carry out upkeep of the lab; support research professors and students; analyze samples; give tours and demonstrations; maintain a safe environment and perform other assigned duties. A bachelor's degree in physical science and/or 4+ years related work experience is required. Candidates should be familiar with electron microscopes, mechanical and electronic equipment, vacuum systems, computers (PC and/or Mac) and EDS/WDS systems. Experience with a microprobe would be especially valuable. Good communication and interpersonal skills are essential.
Lehigh University offers excellent benefits including medical, vision and tuition. Interested candidates should forward their resume to Jennifer Mohney, Human Resources, 428 Brodhead Avenue, Lehigh University, Bethlehem PA 18015. EEO/AA
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
One of our students is trying to stain the DNA of osteoblasts using bisBenzamid, Hoechst no. 33258 trihydrochloride. She is finding conflicting information about the concentration to use and the lethal dose.
I would appreciate it if someone can provide or point us to a protocol they have used successfully. We are primarily a materials lab and do not have a lot of biological reference material at hand.
One of our students is trying to stain the DNA of osteoblasts using bisBenzamid, Hoechst no. 33258 trihydrochloride. She is finding conflicting information about the concentration to use and the lethal dose.
I would appreciate it if someone can provide or point us to a protocol they have used successfully. We are primarily a materials lab and do not have a lot of biological reference material at hand.
I was initially hesitant to introduce a subject that gets periodically posted here, but received so many enthusiastic messages about print and negative scanners that I thought I'd continue the thought. I will be happy to summarize the responses and throw in some opinions of my own. As more and more of us convert to "digital darkrooms" we can share more of our experiences with hardware and software.
I am helping a former traditional photographic media user/computerphobe set up digital imaging capabilities since her university/museum department is closing their darkroom facilities and reassigning personnel (sigh). She has what appears to be a decent budget (until I started pricing the good stuff!). I am proposing she get a fast (733MHz) G4 Mac with maximum (1.5GB) RAM, and she saw and fell in love with the Apple 22" cinema display (as did I when I saw it in person). People seem to like the Agfa T2500 scanner for prints and negatives. A moderate color printer, since she has access to other really good printers in her department. A Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop 6.0, for which I'll train her. Corel Draw for vector graphics?
Additions, subtractions and comments will be welcome. I'll summarize after a reasonable amount of time and we'll see if there is a consensus on the ideal digital darkroom!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
How does your institution handle short term storage in the lab of waste osmium tetroxide. Does it need to be in a fume hood? We keep it in closed glass containers mixed with vegetable oil. Does anyone have a reference regarding whether or not it needs to be stored in a hood? Thank you for any responses.
John, I forgot to mention that we do not use formar coated grids. For better assessment of the renal biopsy we do not mince the sample, either. Please see our Web page for more details. {pathology.lsuhsc.edu/Pathist/dx_home.htlm} click on M.diagnostic service and then on renal biopsy. Our diagnostic em lab continues to send poloroid HRLM pictures and TEM B&W contact prints with each report.
I am a laboratory instructor at Brookdale Community College in Lincroft, NJ who operates a Zeiss 9C TEM which was donated to the college. We are still in the process of being able to make our own grids. Does anyone have some grids that they would be willing to sell or donate to the College? Grids of anything would be greatly appreciated, but basic cellular structures is really what we are looking for as we are a community college and only have first and second year students. Thanks in advance for your help.
I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at:
http://www.agfa.com/scanners/duoscan_HiD.html
Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP injet printers with great success.
Cheers! Csaba
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at:
http://www.agfa.com/scanners/duoscan_HiD.html
Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP injet printers with great success.
Cheers! Csaba
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at:
http://www.agfa.com/scanners/duoscan_HiD.html
Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP injet printers with great success.
Cheers! Csaba
-- ____________________________________________ Csaba Cserhati Univ.of Debrecen / Dept. of Solid State Phys. Hungary tel/fax: 36 52 316073 e-mail: cserhati-at-delfin.klte.hu ____________________________________________
Based on historical precedent we keep used osmium tetroxide in the fridge in a "Kilner" jar (for pickled fruit and veg.), which has a rubber seal.
BTW what ratio of vegetable oil to osmium solution do you use?
Dave
On Thu, 5 Apr 2001 20:30:30 -0500 Michele von Turkovich {mvonturk-at-zoo.uvm.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } How does your institution handle short term storage in the lab of waste } osmium tetroxide. Does it need to be in a fume hood? We keep it in closed } glass containers mixed with vegetable oil. Does anyone have a reference } regarding whether or not it needs to be stored in a hood? Thank you for any } responses. } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
In respose to Tina's post, I have not seen any mention on the list of the scanner I purchased a few weeks ago, the Epson Expression 1640XL. It has 1600dpi optical resolution (scans at a hardware resolution of 1600x3200 dpi) 42 bit color (14 bit gray) and Dmax of 3.6. It is large format, and the transparency adapter comes with a range of negative holders. Has SCSI or USB interfaces with firewire as an optional extra (I use USB on a Win 2000 system). Of course, you pay for what you get - it isn't cheap.
We are only just beginning to learn how best to use all the resolution and bit depth we now have, but I and my users love it!
This is not a comparison, of course (I haven't used the other models) but just to say we are happy with what we have.
Tony.
} } Hi, All- } } A colleague has asked for recommendations for setting up a digital } darkroom (fun to spend someone else's money!). This person would benefit } from a really good scanner that could deal with prints, large format } negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked } into an Agfa Duoscan T2500. Do any of you have an opinion about this or } other suitable scanners? } } I know this subject comes up regularly, but I don't feel bad about } introducing it again, since technology evolves so quickly! } } Mahalo, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
Hysterical precedents aside. The idea of the oil is to absorb all tetroxide vapours and render the substance harmless. Metallic Os is essentially non-toxic, so the treated tetroxide waste should smell of whatever vegetable oil and not emit any of the musky smell emitted by osmium tetroxide. Neither should the treated material be regarded as hazardous waste - but I expect that no safety officer would have the courage to make that declaration. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, April 06, 2001 6:52 PM, Patton, David [SMTP:David.Patton-at-uwe.ac.uk] wrote: } } } } Based on historical precedent we keep used osmium tetroxide } in the fridge in a "Kilner" jar (for pickled fruit and } veg.), which has a rubber seal. } } BTW what ratio of vegetable oil to osmium solution do you } use? } } Dave } } } On Thu, 5 Apr 2001 20:30:30 -0500 Michele von Turkovich } {mvonturk-at-zoo.uvm.edu} wrote: } } } } } } } How does your institution handle short term storage in the lab of waste } } osmium tetroxide. Does it need to be in a fume hood? We keep it in closed } } glass containers mixed with vegetable oil. Does anyone have a reference } } regarding whether or not it needs to be stored in a hood? Thank you for any } } responses. } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } }
between membranes in all cellular components that have two or more membrances and
also seeking "volume and size" of all known subcellular organelles with or without membranes. also seeking " other physical measurement parameters" of cellular components by cell type.. by cell age.. etc. thanks.
Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
Adobe Illustrator or Freehand for vector graphics. Corel Draw is less common and has a proprietary file format that has caused me troubles preparing articles for Microscopy Today. Corel can save in other formats, but people have to use that option. Also, Adobe has educational pricing, and special package prices that bundle full versions of Photoshop and Illustrator.
Phil
} Hi, again- } } I was initially hesitant to introduce a subject that gets periodically } posted here, but received so many enthusiastic messages about print and } negative scanners that I thought I'd continue the thought. I will be happy } to summarize the responses and throw in some opinions of my own. As more } and more of us convert to "digital darkrooms" we can share more of our } experiences with hardware and software. } } I am helping a former traditional photographic media user/computerphobe } set up digital imaging capabilities since her university/museum department } is closing their darkroom facilities and reassigning personnel (sigh). She } has what appears to be a decent budget (until I started pricing the good } stuff!). I am proposing she get a fast (733MHz) G4 Mac with maximum } (1.5GB) RAM, and she saw and fell in love with the Apple 22" cinema } display (as did I when I saw it in person). People seem to like } the Agfa T2500 scanner for prints and negatives. A moderate color printer, } since she has access to other really good printers in her department. A } Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop } 6.0, for which I'll train her. Corel Draw for vector graphics? } } Additions, subtractions and comments will be welcome. I'll summarize after } a reasonable amount of time and we'll see if there is a consensus on the } ideal digital darkroom! } } Mahalo, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
We keep used osmium in small sealed bottles in the hood. Our environmentals services office would rather have **undiluted/unmixed** (no oil) waste for them to neutralize. They have no way of knowing whether all the osmium has been denatured by attachment to oil and would have to treat the whole bottle as osmium waste making for more volume to have to dispose of. Check with your hazardous waste office.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Email: inikolak-at-mred.tuc.gr or eisodos-at-otenet. Name: Irene Nikolakaki
Organization: Technical University of Crete
Education: Graduate College
Location: Chanea, Crete,Greece
Question: Dear sir/madam,
I am a postgraduate student at the Technical University of Crete and I seek information (as detailed as possible) on how enumeration of airborne microorganisms collected on Nuclepore filters is done with the use of epi-fluorescence microscopy.I would be grateful if you could provide me with this infromation or any kind of help as to where I can find it.
I both run the microscope labs here and am a researcher in materials engineering.
First I want to say that I strongly support the use of the digital laboratory. While we still occasionally use film for our highest quality requirements, in general we are fully digital. The use of digital cameras has really expanded our undergraduate teaching laboratories and has sped up our research.
I have found one "dark-side" to a digital imaging laboratory as a lab manager. As the lab manager, I have found that keeping a digital laboratory up-to-date is much more expensive than the film laboratory. When we were only film, we had to repair the film cartridges for our Polaroid PN film (it takes about five minutes) and have the microscopes cleaned about once a year at a cost of about $1k.
The digital lab. is much more expensive time and repair wise. Because we crunch our computers with our image size and storage, it takes more of my time to keep stuff going. All our computers are networked and in addition to work, the students tend to junk up the computers with downloads etc which stops them from working for the image processing work. This requires continual monitoring on my part (in spite of rules against using them for these applications!) In addition, keeping computers that will run the data is expensive. I buy pretty much the best out there, but somehow upgrades are still inevitable. I also have to supply print cartridges,etc. Researchers always supplied their own film and dark room supplies. In addition, I've had to have our cameras repaired numerous times. The cost was high (at least $500) and they stayed gone for up to a month. Finally, some of my cameras are about 3 years old. I can see a degredation in the image quality from when they were purchased. The cameras are much noisier. I see a future of regular replacement of my cameras in addition to the computer upgrades. So while the cost to the researchers is lower (which helps me as a researcher), the cost to the lab itself is higher (which hurts me as a lab manager). I'm working on setting up a fee schedule for this equipment but REALLY hate to have to do it. All of you who do this in a university know how painful it is!
Regarding the camera purchase-in addition to considering the camera resolution and cost, I think you should consider the image transfer. I recommend considering a camera with immediate transfer of the image to the microscope if you have numerous inexperienced users. Being able to focus on the screen is extremely helpful. The image transfer time is also important if you have many images to capture. We do image analysis on numerous images and some of the cameras have about a 30 second transfer time for decent resolution. This would be unbearable for the number of images we collect. I'm not sure how the Nikon Coolpix works but this should be considered by anyone that is purchasing a digital camera.
Good luck!
Robin Griffin UAB
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Thursday, April 05, 2001 6:09 PM To: Microscopy Listserver
Hi, again-
I was initially hesitant to introduce a subject that gets periodically posted here, but received so many enthusiastic messages about print and negative scanners that I thought I'd continue the thought. I will be happy to summarize the responses and throw in some opinions of my own. As more and more of us convert to "digital darkrooms" we can share more of our experiences with hardware and software.
I am helping a former traditional photographic media user/computerphobe set up digital imaging capabilities since her university/museum department is closing their darkroom facilities and reassigning personnel (sigh). She has what appears to be a decent budget (until I started pricing the good stuff!). I am proposing she get a fast (733MHz) G4 Mac with maximum (1.5GB) RAM, and she saw and fell in love with the Apple 22" cinema display (as did I when I saw it in person). People seem to like the Agfa T2500 scanner for prints and negatives. A moderate color printer, since she has access to other really good printers in her department. A Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop 6.0, for which I'll train her. Corel Draw for vector graphics?
Additions, subtractions and comments will be welcome. I'll summarize after a reasonable amount of time and we'll see if there is a consensus on the ideal digital darkroom!
Mahalo, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
Hello, On the September/October issue of Microscopy and Microanalysis, they have a cover picture of a yeast SEM image collected by David Scharf. I was wondering if anyone knew the details of how the sample was prepared. It looks as though the sample was processed directly from the medium it was growing on. I am used to processing bacteria and yeast by filtering through membrane filters, or depositing on polylysine treated glass/silica. I was wondering if there was something different done for this sample?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I am in need of some form of a conductive film coater for SEM sample preparation. I have found a Tousimis Research Corporation Samsputter-2a sputter coater, though it may not be working properly.
Until now, I have been considering constructing my own sputter coater from microwave oven parts and the like, but I have not yet found any books or other resources with clear enough information to get my confidence to the point that I feel that I can do it. I have an understanding of what is needed electrically, but I need something that fills in some details like "Be sure that the target to sample distance can be adjusted between X and Y. You need to get a QRZ type needle valve to admit the argon", etc.
Does anyone have experience with the Tousimis Samsputter 2a? From what I can see, it is an extremely simple unit. Is it something that would be worth repairing? Would it best be used to scavenge parts for a home-built unit?
Since I have breeched the question of home-built sputter coaters, does anyone know of any resources that describe the dos and donts of building one? I know that there are a number of vendors on the listserv. What are the critical features of your units that I just can't get in a home-built unit and just can't live without?
To put this all in perspective, I have an old SEM with 100 Angstrom resolution at best. I don't need flawless, grainless deposition. I have close to zero budget. Even a used sputter coater at market prices is far too expensive. Either I make something like this work, or I learn to work with uncoated samples (which will be difficult, since I am interested in looking at clays - small particle size with next to zero conductivity).
I suggest to use SEM (with calibrated magnification) if you could electroplate your sample with layer of Ni 0.1-0.3 mm thick. Sorry, I do not have a protocol for plating with Ni now, but I did it many times and it is pretty easy.
After coating with Ni you can cut your sample, mount it in epoxy or thermoset and prepare usual cross section. Ni would save the edge of steel layer and measurements will be very simple.
If surface of you samples is flat, you can cut them (better with diamond saw), mount them in epoxy with a steel strip (instead of Ni) attached to steel layer and polish. It is less dependable but still not bad method.
Vladimir Dusevich
-----Original Message----- } From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] Sent: Thursday, April 05, 2001 8:25 AM To: microscopy-at-sparc5.microscopy.com Cc: hban-at-eng.uab.edu
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce.
Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
Osmium waste is toxic and hazardous due to its reactivity and heavy metal characteristics. One of the "urban legends" that abounds is that Osmium mixed with oil is "neutralized" and made safe. It is true that much of it is reduced to metallic Osmium. However in a bizarre incident it was shown in our lab years ago that reduced Osmium is very reactive with strong oxidizers. In the incident some reduced Osmium was accidentally spilled in a sink which had a small amount of Hydrogen Peroxide in the drain. The ensuing yellow gas cloud of newly formed Osmium Tetroxide from the exothermic reaction was very toxic!!!! I mention this only to confirm that Sara Miller is right again as usual and that mixing Osmium waste with other compounds does not make it safe and does increase the volume.
Hi All, I've been tearing hair out all day trying to find a recipe for GMA. In the past, I have used either JB-4 or Historesin (or Technovit) embedding kits, which have clear directions for embedding tissue and polymerizing these resins without UV. I recently ordered a straight GMA embedding kit (because it was much cheaper), and received not one bit of direction as to how I should combine the three components. I did find a protocol on EMS' website for UV polymerization, but would much prefer to use the non-UV method, since it allows tissue to be better oriented. Before I start experimenting, I thought I'd see if anyone out there can help. Thanks again for your help. Bald in Iowa, Kristen Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu
Osmium metal is sometimes reported not to react with air, but other sources report slow reactivity of the metal with air to produce OsO4. It is interesting to note that the name osmium is derived from the Greek "osme" meaning smell, referring to the odour of the metal resulting from osmium tetroxide production at its surface. Presumably neither the finely divided metal nor the dioxide (osmium black) can be trusted not to oxidise to OsO4 in air. What makes osmium tetroxide especially hazardous is the fact that it is very volatile. Unlike OsO4 neither osmium metal nor osmium dioxide are volatile, and provided oxygen can be prevented from reaching them they are therefore relatively innocuous. Surrounding them in oil is probably therefore a good strategy, provided the mixture is still treated as toxic waste.
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401
I think that I have missed part of this message string somewhere, but it sounds like you are starting with a uniform thickness steel layer that has been deposited on a ceramic substrate. I will then make the assumption that you are attempting to alter the thickness of the steel layer across the length of the sample to produce a 'wedge'. I am not sure that this will help, but you could try the wedge polishing technique used for TEM sample preparation, or bevel polishing. I have had success bevel polishing IC's, but not to the degree of accuracy that you require. I know that South Bay technologies makes a pretty good tripod with micrometer levels at all three corners that may help you out. The biggest problem will be in setting up to ensure a good gradient. You should probably try it a few times with 'dummy' samples to get the feel for how aggressive your polish angle is. I hope this helps. Nick Aitken
-----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Friday, April 06, 2001 11:37 AM To: microscopy-at-sparc5.microscopy.com Cc: hban-at-eng.uab.edu
I suggest to use SEM (with calibrated magnification) if you could electroplate your sample with layer of Ni 0.1-0.3 mm thick. Sorry, I do not have a protocol for plating with Ni now, but I did it many times and it is pretty easy.
After coating with Ni you can cut your sample, mount it in epoxy or thermoset and prepare usual cross section. Ni would save the edge of steel layer and measurements will be very simple.
If surface of you samples is flat, you can cut them (better with diamond saw), mount them in epoxy with a steel strip (instead of Ni) attached to steel layer and polish. It is less dependable but still not bad method.
Vladimir Dusevich
-----Original Message----- } From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] Sent: Thursday, April 05, 2001 8:25 AM To: microscopy-at-sparc5.microscopy.com Cc: hban-at-eng.uab.edu
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce.
Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
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Jim Darley wrote: ================================================== Hysterical precedents aside. The idea of the oil is to absorb all tetroxide vapours and render the substance harmless. Metallic Os is essentially non-toxic, so the treated tetroxide waste should smell of whatever vegetable oil and not emit any of the musky smell emitted by osmium tetroxide. Neither should the treated material be regarded as hazardous waste - but I expect that no safety officer would have the courage to make that declaration. ================================================== Jim might be right but there are other points of view on this:
1] I have been led to believe that the vegetable oil reduction of remaining unreduced tetroxide reduces things down to the dioxide, not the metal. The dioxide is a black colloidal solid (the metal is a lighter "bluish gray"). Osmium dioxide is itself relatively innocuous, for example, it is not "regulated" as a dangerous good when shipped, either domestically under US DOT rules or internationally under IATA rules.
2] Osmium is a non-renewable resource and as such, we should all be looking for ways to recycle such materials as opposed to taking them to landfills or in the case of osmium, incineration. I am no environmental "activist" myself, but I think we should all be thinking about such things. The possibility of recycling something however, is intimately connected with the economics of recycling vs. the cost of the purchase of new virgin material. And when the "used" osmium tetroxide containing aqueous liquid is "neutralized" in vegetable oil, for those involved in precious metals recycling, this act essentially kills the economics of recycling.
3] We have in beta testing stage right now an "osmium recycling kit". We are looking for a limited number of laboratories (for now, just in the USA) to participate in our beta testing of this kit. If you are interested in participating in this test, let me know off-line and I will send you the details.
4] In the mean time, I would offer the following advice. Consider using one of the other methods described on this listserver in the past, such as the ones involving KOH as a reducing agent. The reduced material in this state can be recycled economically, but only in large quantities. No one laboratory, in our opinion, could generate enough such material over a reasonable period of time, to make recycling and refining, even of the material is in this state, economical.
5] The one thing you don't ever want to do, at least in the USA, is to declare this as any kind of a "hazardous waste". Once something has been declared to be a hazardous waste, that designation can never (if my understanding of regulations is correct) be reversed, and it forever has to be treated as a waste, and translated, that means it is destined for eventual disposal by either landfill or incineration. I want to be very careful here, it is complicated, this is true so long as we are talking about a RCRA hazardous waste, that is, it meets a listing or characteristic definition. One environmental experts tells me the following: "The reason OsO4 (and OsO2, for that matter) are not regulated as hazardous has nothing to do with their human toxicity (or lack of); it is because osmium, from a regulatory standpoint, is not considered toxic to the environment." He also says it is OK to designate something as a hazardous waste with the intent to recycle it; it simply must be managed as a hazardous waste while it is on-site.
So these containers that are holding reduced material (from the tetroxide) for recycling must be labeled properly, and that would mean something like "osmium dioxide for recycling". I am getting into an area that is not black and white defined, and probably varies from institution to institution in the US, not to mention the variation from country to country. But the important thing is that from a regulatory point of view, what that label says ends up determining the possibilities for the ultimate fate of its contents.
We are striving to find a way to help people transfer, on their environmental "accounting sheets", a material from the column saying "incineration" or "landfill", to the "recylcing" column. And for those who worry about such things, from a legal liability standpoint, there is general recognition that if the material is recycled, there is far less legal liability associated with its disposal than if it is incinerated or sent to landfill. And at the same time, recycling keeps this most valuable nonrenewable resource in the stream of commerce and available for future generations.
Disclaimer: SPI Supplies has developed a kit for recycling osmium. It is not yet commercially available, but is in beta test stage. We also have the obviously ulterior motive of making sure there is osmium tetroxide available for future generations of EM users, otherwise there would be no place for firms like SPI Supplies.......
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
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Different people approach such subjects from rather different angles and they may all be right, but considering their own realities.
A couple of submissions approached the subject from an "absolute safety" point of view. I don't subscribe to that because the amount of tetroxide emitted from gram quantities of metal and some dioxide when surrounded by the tetroxide absorbing oil, would be miniscule. In my opinion, Os waste in vegetable oil is not a substantial hazard in the lab or when dumped. For many, particularly in the USA this is irrelevant, because of strict legal obligations.
There is also an environmental angle. Unfortunately, greatest safety regardless of cost, is at odds with good environmental practice. If a lot of material and energy is used to dispose of a low hazard material, then the total environmental cost may be very high when compared with an "acceptable risk" solution. We should not confuse the ever-increasing demands for greatest safety with "environmentally friendly" practices: they are not synonymous, but often mutually exclusive.
I was pleased to read some of Chuck's submission. Whatever the motivation, recycling of a limited resource is commendable. This has been tried before by turning the waste material back into the tetroxide. I understand that few people still practice the regeneration of osmium. One reason is that the actual fixative solution is poorly defined after reclamation from diverse fixation vehicles.
Chuck's idea of turning the material into metallic osmium is possible. Using precision electrolysis, for instance with SS electrodes at ~400volts, with the plate size determining amperage, the recovered metal could be 99.9% pure. The refiner, however, would charge for assaying and refining and this means another business would need to collect the osmium, resulting in double shipping and double mark-ups. Considering the few grams recoverable, instrument cost and maintenance, this would be a marginal business, but one that the larger labs certainly should consider. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Monday, April 09, 2001 4:45 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } ================================================== } Hysterical precedents aside. The idea of the oil is to absorb all tetroxide } vapours and render the substance harmless. Metallic Os is essentially } non-toxic, so the treated tetroxide waste should smell of whatever } vegetable } oil and not emit any of the musky smell emitted by osmium tetroxide. } Neither should the treated material be regarded as hazardous waste - but I } expect that no safety officer would have the courage to make that } declaration. } ================================================== } Jim might be right but there are other points of view on this: } } 1] I have been led to believe that the vegetable oil reduction of remaining } unreduced tetroxide reduces things down to the dioxide, not the metal. The } dioxide is a black colloidal solid (the metal is a lighter "bluish gray"). } Osmium dioxide is itself relatively innocuous, for example, it is not } "regulated" as a dangerous good when shipped, either domestically under US } DOT rules or internationally under IATA rules. } } 2] Osmium is a non-renewable resource and as such, we should all be looking } for ways to recycle such materials as opposed to taking them to landfills or } in the case of osmium, incineration. I am no environmental "activist" } myself, but I think we should all be thinking about such things. The } possibility of recycling something however, is intimately connected with the } economics of recycling vs. the cost of the purchase of new virgin material. } And when the "used" osmium tetroxide containing aqueous liquid is } "neutralized" in vegetable oil, for those involved in precious metals } recycling, this act essentially kills the economics of recycling. } } 3] We have in beta testing stage right now an "osmium recycling kit". We } are looking for a limited number of laboratories (for now, just in the USA) } to participate in our beta testing of this kit. If you are interested in } participating in this test, let me know off-line and I will send you the } details. } } 4] In the mean time, I would offer the following advice. Consider using } one of the other methods described on this listserver in the past, such as } the ones involving KOH as a reducing agent. The reduced material in this } state can be recycled economically, but only in large quantities. No one } laboratory, in our opinion, could generate enough such material over a } reasonable period of time, to make recycling and refining, even of the } material is in this state, economical. } } 5] The one thing you don't ever want to do, at least in the USA, is to } declare this as any kind of a "hazardous waste". Once something has been } declared to be a hazardous waste, that designation can never (if my } understanding of regulations is correct) be reversed, and it forever has to } be treated as a waste, and translated, that means it is destined for } eventual disposal by either landfill or incineration. I want to be very } careful here, it is complicated, this is true so long as we are talking } about a RCRA hazardous waste, that is, it meets a listing or characteristic } definition. One environmental experts tells me the following: "The reason } OsO4 (and OsO2, for that matter) are not regulated as hazardous has nothing } to do with their human toxicity (or lack of); it is because osmium, from a } regulatory standpoint, is not considered toxic to the environment." He } also says it is OK to designate something as a hazardous waste with the } intent to recycle it; it simply must be managed as a hazardous waste while } it is on-site. } } So these containers that are holding reduced material (from the tetroxide) } for recycling must be labeled properly, and that would mean something like } "osmium dioxide for recycling". I am getting into an area that is not black } and white defined, and probably varies from institution to institution in } the US, not to mention the variation from country to country. But the } important thing is that from a regulatory point of view, what that label } says ends up determining the possibilities for the ultimate fate of its } contents. } } We are striving to find a way to help people transfer, on their } environmental "accounting sheets", a material from the column saying } "incineration" or "landfill", to the "recylcing" column. And for those who } worry about such things, from a legal liability standpoint, there is general } recognition that if the material is recycled, there is far less legal } liability associated with its disposal than if it is incinerated or sent to } landfill. And at the same time, recycling keeps this most valuable } nonrenewable resource in the stream of commerce and available for future } generations. } } Chuck }
There is a proposal for TEM characterisation of cadmium telluride nanoparticles for morphology and size information. I would appreciate information from the list members whether
1. CdTe and related compounds are stable under electron irradiation 2. there is a accelerating voltage limit to be adhered to 3. any other precautions required to ensure microscope safety
My primary microscopy experience is with metallic alloys and ceramics. I have this impression that these compounds are low melting, unstable and likely to sputter onto the pole pieces. Kindly correct and advise me. I use a JEOL 2000 EX II top entry stage operating at 200 kV.
---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
Years ago, I did CdTe/CdS junction. As far as I can remember, both layers were beam sensitive and the layer CdS was more so. I could get fairly good twined CdTe structures as well as its overall morphology, but had difficulty revealing the details in CdS as the available effective observation time was much limited plus the CdS layer was very thin. There are a few ways to get around of the issue, including using a) C-coating, b) small spot size (} 2), and perhaps c) proper keV's (I was using 100keV, the max available voltage for me that time). Good luck!
Chao-Ying Ni Scientist Rodel Inc. USA
-----Original Message----- } From: Divakar R [mailto:divakar-at-igcar.ernet.in] Sent: Monday, April 09, 2001 3:58 AM To: Microscopy (E-mail)
There is a proposal for TEM characterisation of cadmium telluride nanoparticles for morphology and size information. I would appreciate information from the list members whether
1. CdTe and related compounds are stable under electron irradiation 2. there is a accelerating voltage limit to be adhered to 3. any other precautions required to ensure microscope safety
My primary microscopy experience is with metallic alloys and ceramics. I have this impression that these compounds are low melting, unstable and likely to sputter onto the pole pieces. Kindly correct and advise me. I use a JEOL 2000 EX II top entry stage operating at 200 kV.
---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
The tripod polisher may just work. Although your sample is larger than anything I have previously tripoded, it should work if you use a plain L-bracket. First, planarize your sample and the two back feet of the tripod polisher. Then adjust the two back feet to give you the wedge angle you desire (just slightly over 1 degree if my trigonometry for your sample is correct). Likewise, the Multiprep system from Allied High Tech should be able to accept and wedge polish a sample of this size.
If this approach does not work then you may need to use a precision lapping and polishing jig. You can either design your own or you may want to check with South Bay Technology. They make several jigs with precise angular control for polishing crystals. I don’t know however, if they will work with samples this large.
Hope this helps.
Eric Windsor
Disclaimer: I have no financial interest in either South Bay Technology or Allied High Tech Products. I am a satisfied customer of both. Also, there may be other products on the market that will work equally well for preparing this sample.
The opinion expressed is my own and not that of my employer (NIST).
Original Message:
We have a professor here who has a 1 cm x 2.5cm steel layer about 100 um in thickness on a ceramic substrate. The metal layer was sputter deposited onto the ceramic substrate. The ceramic substrate extends past the metal layer. He needs to get a thickness gradient across the steel layer along the 2.5 cm length. He would like to have one end at about 50 microns in thickness and the other at 2 microns with a gradually decreasing thickness gradient. Steps down would be ok although a smooth transition would be better. We have a laser profilometer to measure anything that we produce. Does anyone out there have any ideas how to do this? Would a tripod polisher work? I thought about electropolishing and masking off portions at a time but I worry about what will happen at the interface between the ceramic and the metal. Could we alter a dimpler?
i am interested in a rapid i.e. same day processing schedule, for diagnostic } renal bx. it must be reliable and the cutting qualities good, since we do a } lot of low mag work (250x). thanks in advance. } john
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} i am interested in a rapid i.e. same day processing schedule, for diagnostic {BR} > renal bx. it must be reliable and the cutting qualities good, since we do a {BR} > lot of low mag work (250x). thanks in advance. {BR} > john {BR} {/FONT} {/HTML}
} In respose to Tina's post, I have not seen any } mention on the list of the scanner I purchased } a few weeks ago, the Epson Expression 1640XL. } It has 1600dpi optical resolution (scans at } a hardware resolution of 1600x3200 dpi) 42 bit } color (14 bit gray) and Dmax of 3.6.
I would certainly believe the resolution and the color depth for this scanner is adequate, but if scanning TEM films is an issue, I'd seriously advise measuring the optical density of your films ... I've heard these approach OD} 4 ... which would imply you might consider the dedicated film scanners, e.g., Polaroid 45 Ultra or the new Nikon LS-8000.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
I have been listening to the thread on scanners. Has anyone done tests of how accurate they are in absolute terms for quantitative digitization?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Hello, I'm looking for companies (other than Gatan whom I've already contacted) that sell cryo-stage and cryo-prep. systems for a JEOL 5800LV scanning electron microscope. Thanks for any input! Tracey
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
Has anyone used any of the denatured ethanols as a substitute for absolute ethanol?
We have recently run into some difficulty when needing to reorder 200 proof ethanol, which we use for dehydration and infiltration of samples (primarily many types of paper) prior to embedding in Spurrs epoxy, and for cleaning samples (non paper) and lenses of light microscopes. The chemical company selling the ethanol is insisting that we must have a liquor license before they will ship to us.
Ethanol denatured with a variety of substances is readily available and can be shipped with no licensing requirements. Our concern is that the denaturing agent will leave a detectable residue on lenses, samples, and may cause problems with the polymerization of Spurrs. Rather than obtaining a liquor license, we are considering using one of the 100:5 ethanol: methanol blends. If any of you have had successful or unsuccessful experiences substituting denatured ethanol for absolute in embedding or cleaning protocols, I would appreciate hearing from you.
Teresa Boes Hewlett-Packard Analytical and Development Lab 1000 Circle Blvd Corvallis, OR 97330 541-715-7055 teresa_boes-at-hp.com
Sorry about being late with this thread...Out here we do about 750 renal biopsies last year and all our sections are mounted on 300 mesh uncoated thin bar Gilder grids. We have a Philips 208S at 80kV. We can get a majority of the glomerulus to lie in the grid square to be viewed. Most of our images are shout between 2800X and 14,000X.
Last November we finally obtained the AMT Advantage HR 1K x 1K system and use it exclusively for our EM images... We also do some Neuropathology and Surgical Pathology EM in this lab.
Out here we have rigged up a system that all the Pathologists who need the EM images are setup on a EM users group on the network. I them upload the images from the computer here in the scope room to a directory on our network so the Pathologists can view the images right at their desktop. The renal Pathologist here is thrilled with the system... It cuts down our expenses, and it shortens our turnaround time on specimens from 5 days to about 3 days or less....
Eric A. Rosen Electron Microscopist UCLA Medical Center
==============================
} John: } We do around 500 renal biopsies per year and all the sections are } mounted on 200 mesh uncoated copper grids. We have an 8 year old Hitachi } 7100 and use 60kv. The majority of the glomerulus can be viewed with the } 3-4 serial sections lying randomly across the grid bars. We do not need a } picture of the whole glomerulus, rather most pictures are between 3,000 and } 10,000X. } Dr. Tibor Nadasdy is the renal pathologist and decided last year that } all our renal biopsies would be captured with the digital camera onto a } computer and sent up to him via a network to his computer. So, at the } present time we use very little EM film. He diagnoses each biopsy and } e-mails representative digitized images to the nephrologists. } } } } } Karen L. Jensen, M.S. } Project Manager & Associate Scientist } Electron Microscopy Research Core } -----Original Message----- } } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com } [mailto:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] } Sent: Friday, March 30, 2001 2:20 PM } To: microscopy-at-sparc5.microscopy.com } Subject: renal Em } ok taking a little survey. i am in a diagnostic EM lab. we mount out } sections } on formvar coated slotted grids, so he can shoot pics of the whole } glomerlus. } ok my question. how may of you out there doing diagnostis EM on renals do } this? } john
Tracey VG Microtech make Polaron integrated column-mounted cryo systems including a new system designed for high-resolution work with FEGSEMs Emitech have an off-microscope cryo-prep and transfer system suitable for LTSEM with a conventional tungsten filament or LaB6 SEM BalTec make various cryo transfer and preparation systems which could be used to transfer specimens from e.g. a freeze-etcher to SEM
see http://www.kaker.com/mvd/list.html for addresses and contact numbers for these companies Chris
----- Original Message ----- } From: "Tracey M. Pepper" {tpepper-at-iastate.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, April 09, 2001 6:52 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for a 2" and/or 3" scribe tool for GaN. Does anyone have any experience with a quality new/used equipment vendor who supplied such a tool in the past. I am looking for a fairly new programmable scriber.
Eric, I think that your geometry is off. The Tripod Polisher is about 50 mm long from the sample to the feet and has 1 um divisions. The arctan of 1/50000 gives about 0.001 degrees. This job needs an angle given by the arctan of 48/25000 or 0.11 degrees. Plus you need to stick the thickness at one end at a particular thickness -2 um. It is doable with the TP, but will be difficult. You are correct that they will need to planarize the sample. What I would do is also planarize the holder and mount the parallel-sided sample and then set the height of the feet to the thickness of the sample and then add the amount for the angle. Then I would slowly polish using a low value grit 1 or 3 (perhaps even 1/2) um until I went through to the substrate at one end. You would have your angle and thickness values that went from zero to the desired value and a little thicker. The trick is stopping at the right place and accounting for the wear on the feet. You should be able to watch the facet move towards one end. You can watch the progress using a glass plate and look at the thickness fringes at the facet caused by the unpolished and polished surfaces.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Eric Windsor [mailto:Eric.Windsor-at-nist.gov] } Sent: Monday, April 09, 2001 9:15 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Preparation of a steel wedge on a ceramic substrate } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } ---------. } } } Robin, } } The tripod polisher may just work. Although your sample is } larger than } anything I have previously tripoded, it should work if you use a plain } L-bracket. First, planarize your sample and the two back feet of the } tripod polisher. Then adjust the two back feet to give you } the wedge angle } you desire (just slightly over 1 degree if my trigonometry } for your sample } is correct). Likewise, the Multiprep system from Allied High } Tech should } be able to accept and wedge polish a sample of this size. } } If this approach does not work then you may need to use a } precision lapping } and polishing jig. You can either design your own or you may } want to check } with South Bay Technology. They make several jigs with } precise angular } control for polishing crystals. I don't know however, if they } will work } with samples this large. } } Hope this helps. } } Eric Windsor } } Disclaimer: I have no financial interest in either South Bay } Technology or } Allied High Tech Products. I am a satisfied customer of } both. Also, there } may be other products on the market that will work equally well for } preparing this sample. } } The opinion expressed is my own and not that of my employer (NIST). } } Original Message: } } We have a professor here who has a 1 cm x 2.5cm steel layer } about 100 um in } thickness on a ceramic substrate. The metal layer was sputter } deposited } onto the ceramic substrate. The ceramic substrate extends } past the metal } layer. He needs to get a thickness gradient across the steel } layer along } the 2.5 cm length. He would like to have one end at about 50 } microns in } thickness and the other at 2 microns with a gradually } decreasing thickness } gradient. Steps down would be ok although a smooth transition } would be } better. We have a laser profilometer to measure anything that } we produce. } Does anyone out there have any ideas how to do this? Would a tripod } polisher work? I thought about electropolishing and masking } off portions at } a time but I worry about what will happen at the interface } between the } ceramic and the metal. Could we alter a dimpler? } } } Thanks, } Robin Griffin } UAB } } }
How you can get the licence if you do not SELL the alcohol? Licence is only for selling..
} We have recently run into some difficulty when needing to reorder 200 proof } ethanol, which we use for dehydration and infiltration of samples (primarily } many types of paper) prior to embedding in Spurrs epoxy, and for cleaning } samples (non paper) and lenses of light microscopes. The chemical company } selling the ethanol is insisting that we must have a liquor license before } they will ship to us.
Does anybody have experience (good or bad) with Imaging Plate scanners on TEMs?
PLease reply on or off-line, I will post a summary - preserving anonymity if necessary!
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 or 6125 8525 http://www.anu.edu.au/EMU
Your point is well taken, but in this environment, there is no distinction between purchase, application or consumption.
I'm in California and have not run into this situation as yet. But I would bet that the same myopic rules would apply. I buy ethyl alcohol from Ted Pella in Redding, CA. But I don't know the specific proof of the alcohol (Cat.# 19207). It comes in 200mL bottles and poses no problem when purchased in lots of six or fewer bottles.
gg
At 03:38 PM 4/9/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
How do you define "quantitative digitization?" i.e., what variables are you dealing with in this respect? What are the "absolute terms?"
Anyone else have some ideas about this topic?
gg
At 10:10 AM 4/9/2001, you wrote:
} I have been listening to the thread on scanners. Has anyone done } tests of how accurate they are in absolute terms for quantitative } digitization? } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering & } Center for Transportation Nanotechnology } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu } http://www.numis.nwu.edu http://www.ctn.northwestern.edu
I am a Laboratory Technician currently studying for a Diploma in Occupational Health and Safety. My last study module is an Action Research project which I would like to do on the development of ergonomic microscopes and although there is a lot of information on the problems involved in microscopy and methods to relieve them I was wondering if any research has been done into the effectiveness of ergonomic design for improving microscopy diagnosis, and where I could get any papers on the subject. Thanks. Andy.
Perhaps the reason there was no recipe for the GMA is that you can vary the component ratios alot depending on the tissue you're embedding. There are several reviews from the 60s and 70s on different GMA recipes, at least for embedding plant material. One "standard" mix for plant material is 95% v/v pure GMA, 5% v/v polyethylene glycol, 0.15-1.0% w/v benzoyl peroxide (catalyst). With more catalyst, polymerisation is faster and blocks are harder, but too much produces brittle, bubbly blocks. PEG can be 0-10%, PEG 200 and 400 are commonly used.
Like other methacrylate resins, GMA can be heat polymerised but you need to exclude oxygen. You can either seal the GMA blocks in capsules - gelatin capsules for example - dent the cap to exclude as much air as possible, or cover flat embedding moulds so that they are completely sealed - e.g. use a plastic vial cap as flat mould with another on top to seal. If you have access to a vacuum oven, you can polymerise open moulds under nitrogen.
Time to polymerise depends enormously on the composition of the resin, try 60C overnight for starters if using a fairly "standard" mixture.
good luck, cheers, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Just a minor correction / update. VG Microtech recently changed name to Thermo VG Scientific, address and contact details remain the same, except for the surface science website:
The Polaron range continues to be represented in the US by Energy Beam Sciences.
Best regards,
Mike Wombwell Polaron Range Business Manager Thermo VG Scientific The Birches Industrial Estate Imberhorne Lane East Grinstead West sussex RH19 1UB UK Tel: +44(0)1342310296 (direct line) Tel: +44(0)1342327211 (Switchboard) Fax: +44(0)1342315074 email: mike.wombwell-at-scientific.com Website: http://www.polaron-range.com
E & OE
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: 09 April 2001 22:04 To: Tracey M. Pepper Cc: microscopy-at-sparc5.microscopy.com
Tracey VG Microtech make Polaron integrated column-mounted cryo systems including a new system designed for high-resolution work with FEGSEMs Emitech have an off-microscope cryo-prep and transfer system suitable for LTSEM with a conventional tungsten filament or LaB6 SEM BalTec make various cryo transfer and preparation systems which could be used to transfer specimens from e.g. a freeze-etcher to SEM
see http://www.kaker.com/mvd/list.html for addresses and contact numbers for these companies Chris
----- Original Message ----- } From: "Tracey M. Pepper" {tpepper-at-iastate.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, April 09, 2001 6:52 PM
Richard: Not true in the good old US of A! 200 proof EtOH is considered a controlled substance, and purchase of even small quantities is regulated. In the Histology and EM Labs here, we use a fairly large supply, so there is a company license which allows purchase of a specific amount, and that means if demands go up dramatically there is a real issue of obtaining adequate supplies from time to time. I think that the issue may well have to do with the quantities being purchased. If one buys the odd bottle now and then, there is no hassle. But when usage passes a certain point, then licenses are necessary. Keeps all of the bureaucrats in jobs. As for the license, Teresa, it is mostly just paperwork. Requires you to specify purpose for use, amounts that will be required, etc. If I remember correctly (had to do this myself a _very_ long time ago--now an internal supply room clerk does it), this is an annual process. And, like most labs, since you have to account for every drop of reagent that enters and leaves the lab for EPA, state EPA, OSHA, etc, you will likewise enter the EtOH into that stream, thus showing that you aren't using it for more pleasurable ends.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals NB--personal opinions and experiences only expressed above. On Tue, 10 Apr 2001 08:38:10 +1000, Vr. Richard Bejsak-Colloredo-Mansfeld wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | How you can get the licence if you do not SELL the alcohol? | Licence is only for selling.. | | } We have recently run into some difficulty when needing to reorder 200 | proof | } ethanol, which we use for dehydration and infiltration of samples | (primarily | } many types of paper) prior to embedding in Spurrs epoxy, and for cleaning | } samples (non paper) and lenses of light microscopes. The chemical company | } selling the ethanol is insisting that we must have a liquor license before | } they will ship to us. | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I am interested in freeze drying pieces of murine liver tissue for subsequent embedment in Spurr's epoxy resin. Does anyone have experience with this technique? Specifically I would be interested in times and temperatures. I will be using a turbo freeze dryer from EMS to freeze dry the tissue.
Thanks in advance,
Germaine G. Boucher TEM Lab Pfizer Global Research and Development Groton, CT
Would noise be a good criterion? Say, for a perfectly evenly darkened film (if such a thing existed, or at least even on a scale { { collected pixel size) - what is the value of the noise (standard deviation of pixel value) as a function of film darkness (density)?
This would presumably improve with the time of collection. Thus how "good" your scanner is depends on how you run it or whether it lets you take a slower scan or to average multiple scans. With the exception of drum scanners these devices all use CCD arrays. So what is probably most of interest is the signal to noise ratio as a function of illumination intensity, with everything known about CCD's going into determining this. The maximum density the scanner can handle is just the point at which the noise swamps the signal.
There must be some good literature out there on the sources of noise, optimizing collection (scan) time etc. One article which might be a starting point is:
G. H. Campbell, W. E. King and D. Cohen "Analysis of Experimental Error in High Resolution Electron Micrographs", Microscopy and Microanalysis vol. 3 (1997) p. 451.
This is not very detailed, and treats only the total random noise, i.e. grouping noise arising in collecting the image with that arising from the scanner.
Now, finding a good "Consumer Report" test with hard numbers on commercial models is likely to be a lot harder!
Wharton
} -----Original Message----- } From: Gary Gaugler [SMTP:gary-at-gaugler.com] } Sent: Monday, April 09, 2001 10:37 PM } To: L. D. Marks } Cc: MSA listserver } Subject: Re: Scanners: quantitative accuracy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } How do you define "quantitative digitization?" i.e., what } variables are you dealing with in this respect? What are } the "absolute terms?" } } Anyone else have some ideas about this topic? } } gg } } } At 10:10 AM 4/9/2001, you wrote: } } } I have been listening to the thread on scanners. Has anyone done } } tests of how accurate they are in absolute terms for quantitative } } digitization? } } } } ------------------------------------------------------- } } Laurence Marks } } Department of Materials Science and Engineering & } } Center for Transportation Nanotechnology } } Northwestern University } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } mailto:ldm-at-risc4.numis.nwu.edu } } http://www.numis.nwu.edu http://www.ctn.northwestern.edu }
To the person having trouble getting 200 proof ethanol:
If you are embedding in Spurrs, you do not need ethanol or P.O.. Just dehydrate in E M grade acetone and infiltrate with acetone/Spurrs and you will get good results. Keep it simple, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
I have put together a summary of responses I got from my inquiry about a week or so ago about EDS systems. I really appreciate everyone's input. We haven't made any decisions yet since we are still gathering information but your responses will help. I've included the responses in their entirety so I hope this helps others as well.
Thanks again from everyone who contributed.
Jean Ross Central Microscopy Research Facility University of Iowa
----------------------------------------------------------------------------- I have been using an IXRF EDS / Gresham Detector (with IXRF digital pulse processor) system for almost 2 years. Initially, the IXRF was installed on an ETEC. About 15 months ago the Etec was replaced with a Hitachi 3500, a new detector (Gresham) was purchased, and the EDS was installed on the 3500. Generally, I am quite satisfied. There are minor software bugs, but IXRF has been reasonably good at fixing them when discovered. It has been my experience that all the systems have bugs, perhaps some more than others. Prior to the IXRF, I had a Kevex 8000/Delta.
Low end noise and broad peaks were evident on first installation, but were soon fixed by tweaking the detector preamp and pulse processor amp.
I am still running their first software package, "Iridium". I have the newest release, "EDS 2000", but lack of time has kept me from installing and checking it out.
I should mention that IXRF is a "virtual" company, with people spread out between Texas, California, etc. This has not proven to be a problem.
Woody White McDermott Technology, Inc. nwwhite-at-mcdermott.com
We have had EDAX for about a decade and a half, and we are very pleased with the product and the service we get. Carol Heckman heckman-at-bgnet.bgsu.edu
I would recommend you give consideration to Doug Connors at TN Analyzer Service, Inc. of Dane, WI. Doug has rebuild and upgraded detectors for me for the last 6 years. He is dependable knowledgeable, and economical as well.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 (480) 967-3946 bobrobs-at-earthlink.net
We recently purchased a Noran Instruments Vantage DS1. We purchased this based on some very impressive demonstrations of software that the salesperson brought in. Unfortunately they are still working out the bugs in their software. Everything they have is ported over from Unix, and literally runs in a unix shell on an Microsoft Windows NT platform. This makes their software fairly buggy. Their response time to fix major bugs and hang-ups in the software has been very slow, and if given the opportunity to do it all over again I'd probably look at Oxford Instruments. I would still rate the quality of the equipment very high. Our detector performs at the specified resolution, and is a good piece of equipment. Now if they could only get the software end of it straight... My vote: 1) Oxford Instruments 2) Noran Instruments 3) Edax or some of the smaller players The benefit with going with a larger company is support and upgrades. We have a 10 year old WDX that we just purchased new software and interface for last year. Our old EDS was given some trade in value by Noran. And we all know how valuable a service contract can be...
Get back to me if you have any more questions, ~Jonathan Jonathan Dunlap Analytical Laboratory Manager Osram Sylvania Inc. 816 Lexington Avenue Warren, PA 16365 Ph: 814-726-6991 Fax: 814-726-6942 Jonathan.Dunlap-at-sylvania.com
We have an Oxford Instruments Link ISIS Model 200 on our 2460N. We have been happy with it, but I don't know about the direction that Oxford is heading. I don't care for the feel of their new INCA software. Some might like it. It also seems to be slow coming together. Some of the functions are still lacking after 2 (or is it 3) years of seeing it at MSA.
We have an IXRF system on our JEOL 840A. It was a good price ($30K) for an upgrade to our Kevex several years ago. It does what we need. They keep at work on the software and have it freely available on the web. I might have to pay closer attention and stay away from the beta stuff. They are still working on it. They also have a nice digital pulse processor which stills stand alone for about $5k.
I still feel funny about some contacts with EVEX. I can't say much about EDAX, NORAN, or PGT. They should all have good stuff but it might be pricey. The last we seriously looked at them was 6 years ago or so when we opted for the Oxford.
I was intrigued by the unit from Quartz PCI. I think it was called X-ray One, or such. It was new at MSA 1-1/2 years ago but looked promising.
We purchased an EDAX Falcon system for our Hitachi S-3000N and I've been pleased with it. It has better light element sensitivity than most which was very important to me although I don't think that its mapping capabilities are as good as PGT's, say. I don't have direct experience with Noran although I did talk to them and their system seemed ok - but logistics didn't favor Noran so I passed on them. EDAX does have good integration with the Hitachi and the Quartz database.
I'd be glad to respond more specifically if you'd like.
Richard Shalvoy Arch Chemicals Cheshire, CT RBShalvoy-at-archchemicals.com
I have an iXRF systems out of Texas using a Gresham detector. It works well. Not the most cutting edge, but they are one of the "start ups". They have been around for I guess 6-7 years. I have a digital pulse processor and completely active control for x-ray maps and such. They are very price competitive, but lack a dedicated technical support person. You talk to the programmer or electronics expert, but no techs on the phone whenever you have a software question. But, if you willing to wait a day for some answers then they are worth it. I haven't run across the problem where I thought, "if I just had a better system". If you want to integrated w/ WDS than maybe Noran. Also, if you want to integrated w/ motorized stage control, I don't think they off such a package, like the bigger companies.
I have a Hitachi 450. I used to run a 2400 and 500 before I quit my day job and went out on my own. I am very happy w/ Hitachi.
I use Oxford ISIS300 system on HITACHI S-3500N (with VP mode) for light element analysis, mostly C, O, N, F, P, S, Si, Mg, as well as metal Co, Ni-P, Pt, Cr, Fe, W, etc. This system works well. One useful function is the overlay of 2 spectrums. I can easily subtract the blank from the sample spot and make it easy to identify what is (are) in the sample. I am sure some other program may have this kind of function, but I have not seen.
Zhiyu Wang zhiyuw-at-home.com I would be interested in seeing the responses as I am going to try and get funding next year for a replacement for our EDAX PV9100 on an Hitachi s-450.
We have been running EDX on SEMs and TEMs for many years. We used to have a range of systems from Kevex, PGT, Noran, Link, EDAX, however a few years ago we decided that we ought to standardize on one common system. After evaluation we bought three Oxford Instruments ISIS systems. Whenever we have upgraded or bought new systems they have been Oxford Instruments ISIS or now INCA.
I have been happy with the ISIS except for the file handling that was not designed for a multi user facility such as ours (approx 120 EM users in total roughly 25 to 30 swapping every year). I am really quite impressed by the INCA, Oxford Instruments are, at last, listening to the users and adding user requested facilities. They have sorted out the file handling mess of the ISIS and structured it well for an SEM user (not quite as well for a TEM user but there are less of us). The software structure is quite intuitive and there is a really impressive help menu and explanation of everything from the physics of X-ray generation, how EM’s work, how detectors work and how to analyze samples.
Their detectors have always been good and the SATW (thin window detectors) still have a reasonable efficiency at low Z. B is possible but C is easy and even the N peak is over 30% efficient (there is often a high absorption at N).
Another feature that is invaluable for TEM is the integral shutter that will close when the count rate is too high. This protects the crystal, it prevents it overloading and shutting down or worse the crystal efficiency may change for a few minutes until it recovers fully. This may affects your quantitative work. In TEM this is usually caused by hitting the grid bar and not really a problem in SEM but I don't know what secondaries and ions you will have in a variable pressure SEM. It could be useful for you, check with other high pressure SEM users.
Regards, Ron
Please note: Oxford Instruments have upgraded an ISIS to an INCA system in my department, without charge, in return for access to the instrument for development projects and demonstrations for a fixed number of days. I receive no benefit from this and the department has no benefit from Oxford Instruments sales. I remain a thorn in the flesh of all our suppliers if I think they could improve their products or service. ron.doole-at-materials.oxford.ac.uk
Hello, I am very familiar with the Oxford ISIS 300 series spectrometers. They are ok, and the new Inca system looks good too. However, I recently saw the PGT spectrometer at Lehigh and it is very impressive.
I'm also in the market for an EDS system and have looked at EDAX, Noran, PGT and Oxford.
I edited out PGT because in order to quantify you have to optimize the system for the type of sample by playing with fudge factors, which none of the other systems have to do (though one of them, I think Noran, lets you adjust a sensitivity factor if you want to, but they didn't do it on my samples that were tested against known microprobe results and the answers were fine). I also eliminated Oxford, though it has a terrific user interface (maybe at the expense of functionality), because they consistently IDed my aluminum peaks as Br or Tm (!); this made me wonder about all their algorithms. They claim it had to do with the takeoff angle on the particular SEM being used, but that shouldn't be a factor.
I like both EDAX and Noran, though for different reasons. EDAX user interface is better than Noran's, though again, I think Noran possibly offers more routines (it's hard to keep track and see absolutely everything a system has to offer in a demo day....).Noran can multitask - work on several programs while a spectral map is being collected, for instance (does EDAX? I have to check). But EDAX has a beam skirt reduction routine for low vac mode (though it's time consuming, so a bit cumbersome), and their peak modeling is right up front - but Noran can put theirs up front also if you want to have it accessible (yes) and I think Noran might be a little better engineered.
As you can see I'm still in a quandary (ditto for the two contending SEMs, LEO and ESEM). Whatever I decide I'll still be very interested in the results of your posting - especially if other folks' info comes in within the next week or so it would help in my decision too.
I hope my input helps a little. Good luck with your quest!
I looked into Noran, EDAX, and PGT. Noran was quickly culled (less user friendly, less abilities, didn't work right during demo), but EDAX and PGT both seemed to have equivalent capabilities (PGT claimed a 'proprietary' signal amplifier/digitizer doohickey, but it was only in the placement.) For the long-time spectrum gathering (I forget the technical term), PGT makes many passes with short dwell times while EDAX dwells on each pixel much longer to collect data & does it in 1 pass. Kinda 6 of one, half dozen of the other. What made us choose the EDAX Phoenix system was the fact that the PGT software was UNIX-based (although hidden) while EDAX is PC-based. I've heard rumors that PGT is switching to PC-based; we purchased our system in 1999. I've also heard that Noran has practically no service techs (but that may be an East coast thing.) We've been happy with the EDAX service, and I enjoyed their user school very much. By the way, our SEM is a variable pressure JEOL 5900, and it's integrated with the EDAX system.
Hope I've been helpful,
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
Hi All: I have been asked to examine some films on a Si carbide substrate using TEM. Does anyone out there work with this material. Can one mechanically thin it using diamond films? Please offer some suggestions of what you are doing. Thanks in advance, Michael Coviello University of Texas Arlington
We have some staining that suggests the staining dye has transferred from one place to another and I write to see if anyone could shed some light on this for us.
In the experiment we stain isolated chromosomes with DAPI, rinse them, and then introduce them into plant protoplast cells (likely one or few per cell). There is no autofluroescence in the DAPI channel and we can follow the course of the dyed chromosomes over time. At first the DAPI fluorescence appears in the cytosol on structures likely to be the introduced chromosomes. Then it appears in the nucleus, where all the chromosomes are stained! This is especially evident when we culture these cells. Dividing cells at metaphase show DAPI fluorescence over the entire metaphase plate. Note that only some cells show DAPI fluorescence, consistent with the presumption that our fusion process that introduces the chromosomes is only successful in some of the cells.
Has anyone had an experience where dye has been transferred from one structure to another in a living cell? What are some alternative interpretations of this phenomenon?
Thanks for your comments,
Howard Berg
R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility, Associate Member Donald Danforth Plant Science Center/Nidus Center 893 North Warson St. Louis, MO 63141
We tripod polished thin films on SiC. It works OK with diamond lapping film and diamond powder slurry as a final polish, but it takes a loooonnng time. Put a sacrificial piece of SiC on the second side tool pedestal so the final wedge polish will go from SiC sample to SiC tool. Otherwise the glass pedestal polishes faster than the SiC and makes a little step that can lead to sample breakage. ...Always a good idea to match the pedestal hardness to the specimen--hard or soft.
As I recall, we had more problems with poor film adhesion to the SiC than with polishing the SiC substrate.
Although we haven't tried it, I suppose a FIB would work fine.
Ron
There was a thread recently on scanners for TEM film. I have looked up all the models mentioned, on the web and called agents for prices - and produced a comparative table, given below.
I do not guarantee that the figures are accurate but they are my best interpretation of the data given.
In the light of experience and Nestor's comments, I would suggest that 2000 dpi is a minimum for TEM negatives. You may be able to get away with less nine times out of ten, but there will be occasions when you need more. I would exclude the Minolta and all the Epsons from consideration (despite the incredibly low prices of some of the Epsons) because of the low pixel density.
Among the rest the Nikon has the best pixel density and the best optical density (another critical parameter for TEM negatives). The price is very competitive too. The Nikon web site does not give a time for scanning a negative. On the face of it the Nikon would be a best buy - get a separate, inexpensive flatbed scanner for the other work.
These comments are all my own opinions based on manufacturers' data. Since we are considering purchase any comments to the contrary would be most welcome.
Code Maker Model Type
A Agfa DuoScan T2500 Flatbed -Transparency included
B Epson 1640 several versions Flatbed -Transparency option 1680 several versions
C 1600 several versions Flatbed -Transparency included
D Imacon Flextight Precision II Drum -for film and large format
E Minolta Dimage ScanMulti II Film
F Nikon Super Coolscan 8000ED Film
G Polaroid 45 Ultra Film
H Umax Powerlook 3000 Flatbed -Transparency included
Code dpi OD Time Price Opinion at 6 x 9 cm
A 2500 x2500 3.4 3 min $4,500 Fair
B 1600 x 3200 3.6 $300-$3000 Poor $800-$1400 Poor
C 1600 x 3200 3.3 $650-$1160 Not suitable
D 2240 x2240** 3.9/4.1 N/A above $10k Good: low pixel density
E 1128 x 1128 3.6 Not suitable
F 4000 x 4000 4.2 N/A $2,695 V. Good
G 2500 x 2500 3.8 5 min $7,495 Good but pricey
H 3048 x 3048 3.6 3 min $6,499 Good
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I too am about to buy and I would make a couple of comments on your evaluation. First, let me remind everyone that the Dynamic range is a log scale so small numerical differences are significant.
I also think the Nikon Coolscan 8000 looks great but it only takes a 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone to a smaller film size or is Nikon using a non-Japanese EM as their standard? seems odd but I don't see how the Nikon would be very useful. You say a {2000 line scanner would be useful 9 out of 10 times but want the 2000+ lines for the occasional high res scan. I would argue that the size of the negative was the more important variable to be worried about. The Nikon couldn't handle 4x5 LM negatives or transparencies from autoradiography of Westerns/Northerns, etc.
My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. more details at www.microtek.com. This is my leading candidate. It was 4 negative carriers and I await word whether one could be modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have my scientific instrumentation shop guys fabricate a holder. It comes with a glass 8 x 10 glass carrier for odd size negs but I want to avoid Newton rings and want a glassless carrier.
I would appreciate comments on the following argument (I think I have this correctly figured out but am not sure since so many out there seem to want to have a higher resolution scanner). I have a Fuji Pictrography 3000 printer with a 400 dpi output that is as good as any other widely available printer in the academic world. If you figure the maximum published image size is about 8 inches, that would mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% depending on the orientation of the negative and still be taking full advantage of the printer resolution. In reality, most EM publication prints are smaller than 8" wide so one could crop even more and still not need more than 1000 dpi. A resolution } 1000 dpi would be useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x 4 negative would be 192 MB. That is pretty big for doing morphometry on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB and that is much more manageable. Perhaps the difference is in the type of EM we are doing. I am working with biological specimens doing standard thin section type stuff. are you doing some Material Sci application that demands more?
I will be interested in Alwyn (and any others) reply since I hope to buy one soon!
} . } } } There was a thread recently on scanners for TEM film. I have looked up } all the models mentioned, on the web and called agents for prices - and } produced a comparative table, given below. } } I do not guarantee that the figures are accurate but they are my best } interpretation of the data given. } } In the light of experience and Nestor's comments, I would suggest that } 2000 dpi is a minimum for TEM negatives. You may be able to get away } with less nine times out of ten, but there will be occasions when you } need more. } I would exclude the Minolta and all the Epsons from consideration } (despite the incredibly low prices of some of the Epsons) because of the } low pixel density. } } Among the rest the Nikon has the best pixel density and the best optical } density (another critical parameter for TEM negatives). The price is } very competitive too. The Nikon web site does not give a time for } scanning a negative. On the face of it the Nikon would be a best buy - } get a separate, inexpensive flatbed scanner for the other work. } } These comments are all my own opinions based on manufacturers' data. } Since we are considering purchase any comments to the contrary would be } most welcome. } } } } Code Maker Model Type } } } } A Agfa DuoScan T2500 Flatbed } -Transparency included } } B Epson 1640 several versions Flatbed } -Transparency option } 1680 several versions } } C 1600 several versions Flatbed } -Transparency included } } D Imacon Flextight Precision II Drum -for } film and large format } } E Minolta Dimage ScanMulti II Film } } F Nikon Super Coolscan 8000ED Film } } G Polaroid 45 Ultra Film } } H Umax Powerlook 3000 Flatbed } -Transparency included } } } } } } Code dpi OD Time Price } Opinion } at 6 x 9 cm } } } A 2500 x2500 3.4 3 min } $4,500 Fair } } B 1600 x 3200 3.6 } $300-$3000 Poor } } $800-$1400 Poor } } C 1600 x 3200 3.3 } $650-$1160 Not suitable } } D 2240 x2240** 3.9/4.1 N/A above } $10k Good: low pixel density } } E 1128 x 1128 3.6 } Not suitable } } F 4000 x 4000 4.2 N/A } $2,695 V. Good } } G 2500 x 2500 3.8 5 min } $7,495 Good but pricey } } H 3048 x 3048 3.6 3 min } $6,499 Good } } } -- } .......... } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvania 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I assume that your SiC is single crystal. The small angle cleavage technique works for SiC. See MRS Proceedings, TEM Prep IV Vol 480.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Michael Coviello [mailto:coviello-at-mae.uta.edu] } Sent: Tuesday, April 10, 2001 10:59 AM } To: listserver } Subject: TEM-SiC wafer sample prep? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Hi All: } I have been asked to examine some films on a Si carbide } substrate using } TEM. Does anyone out there work with this material. Can one } mechanically thin it using diamond films? Please offer some } suggestions } of what you are doing. } Thanks in advance, } Michael Coviello } University of Texas Arlington } } }
Microscopy Scientist for two Recently Established NIH Centers for Biomedical Research at The University of Wyoming
As part of our seven interrelated projects into the biology, chemistry, and molecular biology of cardiovascular function and nitric oxide we seek a skilled microscopist to be hired at the non-tenured Assistant Research Professor level. Experience and research interests in state-of-the-art microscopy, including confocal and epifluorescence, and ultrastructural techniques. Primary responsibility will be management of the University's Microscopy Center. Opportunity includes the possibility of establishing a research program within this Center. Appointment will be at a (non-tenure track) Assistant Professor level for a renewable four year term.
Job Description: The person we seek will be responsible for organization of a new research laboratory facility at the University of Wyoming which will include two confocal microscopes and an electron microscope. The position includes overall management of the microscope facility, user training, and user supervision. Requirements for the position include experience with light, confocal, and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, and record keeping. The individual will work with only minimum supervision. Responsibilities include: Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility. In addition the manager will perform preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing. Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication. Applicant Qualifications: Experience focus on both confocal and EM. Regarding confocal microscopy, we require experience with confocal and digital imaging techniques, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization. The successful applicant will have experience with tissue preparation for EM and the maintenance of an electron microscope. Excellent interpersonal and organizational skills are essential. Ph.D.. degree required Desirable Experience: Expertise and training in the operation of confocal microscope and EM microscope systems is required. Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are desirable. Significant facility with computers is desired. Salary Range: Commensurate with experience.
For additional information see our websites: www.uwyo.edu/nocobre www.uwyo.edu/MolecBio/Cobre To apply send complete CV, three references, and a cover letter indicating which position(s) you are applying for to: Lynda Payne, Department of Chemistry, University of Wyoming, Laramie, WY 82071-3838, USA. The searches will remain open until all positions are filled.
The University of Wyoming is located in a high (2,200 m) valley surrounded by the Rocky Mountains in the southeast corner of Wyoming. The University of Wyoming is an equal opportunity/affirmative action employer.
The Beam, newsletter for the Southeastern Microscopy Society (SEMS) is now online at the SEMS website in PDF format: http://www.biotech.ufl.edu/sems/
It contains information on the upcoming meeting in Clemson, SC. If you are a member, and have not received this notice via e-mail, and wish to be informed about the society through e-mail, please respond off-listserve at: jshields-at-cb.uga.edu
John Shields Center for Ultrastructural Research Univ. of Georgia Athens, GA
Such an instrument could provide for low-angle rotary shadowing capability for visualizing purified proteins at high resolution. It would need to have an electron beam gun for platinum evaporation and a separate one for carbon coating.
If this is what you have in mind, I support it. I purchased such an instrument that had good performance for about $20,000 about 10 years ago.
Dr. Joseph C. Besharse Professor and Chairman Dept of Cell Biology, Neurobiology and Anatomy Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, WI 53226-0509
} From: Gang Ning {gning-at-mcw.edu} } Organization: Medical College of Wisconsin } Reply-To: gning-at-mcw.edu } Date: Tue, 10 Apr 2001 14:17:42 -0500 } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } Cc: "Dr. Traktman" {ptrakt-at-post.its.mcw.edu} , Ming Lei {mlei-at-mcw.edu} , "Dr. } Besharse" {jbeshars-at-mcw.edu} } Subject: Sputter coater } } Hi All: } } I want to buy a new/used sputter coater which enables to do rotary } shadowing as well as carbon coating. Any suggestions/input are } appreciated. } } Greg Ning } } EM Facility } Medical College of Wisconsin }
Question: Hi, I have just purchased a microscope, a good biological one. I need a microscope kit but nobody sells them around here. Anyway already I have glass slides and covers. I have read some on microscopy and I need an adhesive, resin I think its called to prepare slides? is this true? Also some ink to stain specimens. What are the names of all these chemicals so I can buy them all seperatly since no one sells them all together. Thankyou.
Dear Theresa: Although a metallographic laboratory, we use alcohol for specimen preparation, mainly for preparation of etchants and for specimen cleaning. We found the paperwork associated with pure ethanol onerous and switched to denatured alcohol Type 3A with no discernable difference. Beware of some of the denaturants, they produce unusual side effects.
Sam Purdy National Steel Tech Center Trenton MI
} ---------- } From: BOES,TERESA (HP-Corvallis,ex1) } Sent: Monday, April 9, 2001 2:50 PM } To: 'microscopy-at-MSA.microscopy.com' } Subject: Substitutes for absolute ethanol? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Has anyone used any of the denatured ethanols as a substitute for absolute } ethanol? } } We have recently run into some difficulty when needing to reorder 200 } proof } ethanol, which we use for dehydration and infiltration of samples } (primarily } many types of paper) prior to embedding in Spurrs epoxy, and for cleaning } samples (non paper) and lenses of light microscopes. The chemical company } selling the ethanol is insisting that we must have a liquor license before } they will ship to us. } } Ethanol denatured with a variety of substances is readily available and } can } be shipped with no licensing requirements. Our concern is that the } denaturing agent will leave a detectable residue on lenses, samples, and } may } cause problems with the polymerization of Spurrs. Rather than obtaining a } liquor license, we are considering using one of the 100:5 ethanol: } methanol } blends. If any of you have had successful or unsuccessful experiences } substituting denatured ethanol for absolute in embedding or cleaning } protocols, I would appreciate hearing from you. } } Teresa Boes } Hewlett-Packard } Analytical and Development Lab } 1000 Circle Blvd } Corvallis, OR 97330 } 541-715-7055 } teresa_boes-at-hp.com } }
I would caution biased opinions from installation engineer and sales representatives ie... James Fotinopoulos.... www.semguy.com...
Hmmmmmmmmm?
Food for thought
----- Original Message -----
} From: "jeanross" {jeanross-at-blue.weeg.uiowa.edu} } To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, April 10, 2001 10:22 AM } Subject: EDS summary } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I have put together a summary of responses I got from my inquiry about a } week } } or so ago about EDS systems. I really appreciate everyone's input. We } } haven't made any decisions yet since we are still gathering information } but } } your responses will help. I've included the responses in their entirety } so I } } hope this helps others as well. } } } } Thanks again from everyone who contributed. } } } } Jean Ross } } Central Microscopy Research Facility } } University of Iowa } } } } -------------------------------------------------------------------------- } --- } } I have been using an IXRF EDS / Gresham Detector (with IXRF digital pulse } } processor) system for almost 2 years. Initially, the IXRF was installed on } } an ETEC. About 15 months ago the Etec was replaced with a Hitachi 3500, a } } new detector (Gresham) was purchased, and the EDS was installed on the } 3500. } } Generally, I am quite satisfied. There are minor software bugs, but IXRF } } has been reasonably good at fixing them when discovered. It has been my } } experience that all the systems have bugs, perhaps some more than others. } } Prior to the IXRF, I had a Kevex 8000/Delta. } } } } Low end noise and broad peaks were evident on first installation, but were } } soon fixed by tweaking the detector preamp and pulse processor amp. } } } } I am still running their first software package, "Iridium". I have the } } newest release, "EDS 2000", but lack of time has kept me from installing } and } } checking it out. } } } } I should mention that IXRF is a "virtual" company, with people spread out } } between Texas, California, etc. This has not proven to be a problem. } } } } Woody White } } McDermott Technology, Inc. } } nwwhite-at-mcdermott.com } } } } -------------------------------------------------------------------------- } --- } } } } } } We have had EDAX for about a decade and a half, and we are very pleased } with } } the product and the service we get. } } Carol Heckman } } heckman-at-bgnet.bgsu.edu } } } } -------------------------------------------------------------------------- } --- } } } } } } look at IXRF eds systems there web site is www.ixrfsystems.com, they are } } very affordable and offer no nonsense performance that second to none. } } } } happy ixrf user, } } } } James Fotinopoulos } } yzfrjim-at-ix.netcom.com } } } } } } -------------------------------------------------------------------------- } ---- } } } } I would recommend you give consideration to Doug Connors at } } TN Analyzer Service, Inc. of Dane, WI. Doug has rebuild and } } upgraded detectors for me for the last 6 years. He is dependable } } knowledgeable, and economical as well. } } } } Bob Roberts } } EM Lab Services, Inc. } } 2409 S. Rural Rd Suite C } } Tempe, Arizona 85282 } } (480) 967-3946 } } bobrobs-at-earthlink.net } } } } -------------------------------------------------------------------------- } --- } } } } } } We recently purchased a Noran Instruments Vantage DS1. We purchased this } } based on some very impressive demonstrations of software that the } salesperson } } brought in. Unfortunately they are still working out the bugs in their } } software. Everything they have is ported over from Unix, and literally } runs } } in a unix shell on an Microsoft Windows NT platform. This makes their } } software fairly buggy. Their response time to fix major bugs and hang-ups } in } } the software has been very slow, and if given the opportunity to do it all } } over again I'd probably look at Oxford Instruments. I would still rate } the } } quality of the equipment very high. Our detector performs at the } specified } } resolution, and is a good piece of equipment. Now if they could only get } the } } software end of it straight... } } My vote: } } 1) Oxford Instruments } } 2) Noran Instruments } } 3) Edax or some of the smaller players } } The benefit with going with a larger company is support and upgrades. We } have } } a 10 year old WDX that we just purchased new software and interface for } last } } year. Our old EDS was given some trade in value by Noran. And we all } know } } how valuable a service contract can be... } } } } Get back to me if you have any more questions, } } ~Jonathan } } Jonathan Dunlap } } Analytical Laboratory Manager } } Osram Sylvania Inc. } } 816 Lexington Avenue } } Warren, PA 16365 } } Ph: 814-726-6991 } } Fax: 814-726-6942 } } Jonathan.Dunlap-at-sylvania.com } } } } } } -------------------------------------------------------------------------- } ---- } } } } } } We have an Oxford Instruments Link ISIS Model 200 on our 2460N. We have } been } } happy with it, but I don't know about the direction that Oxford is } heading. I } } don't care for the feel of their new INCA software. Some might like it. It } } also seems to be slow coming together. Some of the functions are still } lacking } } after 2 (or is it 3) years of seeing it at MSA. } } } } We have an IXRF system on our JEOL 840A. It was a good price ($30K) for an } } upgrade to our Kevex several years ago. It does what we need. They keep at } } work on the software and have it freely available on the web. I might have } to } } pay closer attention and stay away from the beta stuff. They are still } working } } on it. They also have a nice digital pulse processor which stills stand } alone } } for about $5k. } } } } I still feel funny about some contacts with EVEX. I can't say much about } EDAX, } } NORAN, or PGT. They should all have good stuff but it might be pricey. The } } last we seriously looked at them was 6 years ago or so when we opted for } the } } Oxford. } } } } I was intrigued by the unit from Quartz PCI. I think it was called X-ray } One, } } or such. It was new at MSA 1-1/2 years ago but looked promising. } } } } Feel free to call if you want more details. } } } } Warren E Straszheim } } wesaia-at-iastate.edu } } } } -------------------------------------------------------------------------- } --- } } } } } } We purchased an EDAX Falcon system for our Hitachi S-3000N and I've been } } pleased with it. It has better light element sensitivity than most which } } was very important to me although I don't think that its mapping } } capabilities are as good as PGT's, say. I don't have direct experience } with } } Noran although I did talk to them and their system seemed ok - but } logistics } } didn't favor Noran so I passed on them. EDAX does have good integration } } with the Hitachi and the Quartz database. } } } } I'd be glad to respond more specifically if you'd like. } } } } Richard Shalvoy } } Arch Chemicals } } Cheshire, CT } } RBShalvoy-at-archchemicals.com } } } } -------------------------------------------------------------------------- } --- } } } } } } I have an iXRF systems out of Texas using a Gresham detector. It works } } well. Not the most cutting edge, but they are one of the "start ups". They } } have been around for I guess 6-7 years. I have a digital pulse processor } } and completely active control for x-ray maps and such. They are very price } } competitive, but lack a dedicated technical support person. You talk to } the } } programmer or electronics expert, but no techs on the phone whenever you } } have a software question. But, if you willing to wait a day for some } } answers then they are worth it. I haven't run across the problem where I } } thought, "if I just had a better system". If you want to integrated w/ WDS } } than maybe Noran. Also, if you want to integrated w/ motorized stage } } control, I don't think they off such a package, like the bigger companies. } } } } I have a Hitachi 450. I used to run a 2400 and 500 before I quit my day } job } } and went out on my own. I am very happy w/ Hitachi. } } } } Good Luck } } } } Fell free to call with any specifics. } } } } Their web page is www.ixrfsystems.com } } } } Ric } } } } SMARTech } } 860-491-3299 } } www.semguy.com } } 19 Cornwall Drive } } Goshen CT 06756 } } smartech-at-javanet.com } } } } -------------------------------------------------------------------------- } ---- } } } } } } Hello, all: } } } } I use Oxford ISIS300 system on HITACHI S-3500N (with VP mode) for light } } element analysis, mostly C, O, N, F, P, S, Si, Mg, as well as metal Co, } Ni-P, } } Pt, Cr, Fe, W, etc. This system works well. One useful function is the } overlay } } of 2 spectrums. I can easily subtract the blank from the sample spot and } make } } it easy to identify what is (are) in the sample. I am sure some other } program } } may have this kind of function, but I have not seen. } } } } Zhiyu Wang } } zhiyuw-at-home.com } } I would be interested in seeing the responses as I am going to try and get } } funding next year for a replacement for our EDAX PV9100 on an Hitachi } s-450. } } } } Dave } } David.Patton-at-uwe.ac.uk } } } } -------------------------------------------------------------------------- } --- } } } } } } We have been running EDX on SEMs and TEMs for many years. We used to have } a } } range of systems from Kevex, PGT, Noran, Link, EDAX, however a few years } ago } } we decided that we ought to standardize on one common system. After } evaluation } } we bought three Oxford Instruments ISIS systems. Whenever we have upgraded } or } } bought new systems they have been Oxford Instruments ISIS or now INCA. } } } } I have been happy with the ISIS except for the file handling that was not } } designed for a multi user facility such as ours (approx 120 EM users in } total } } roughly 25 to 30 swapping every year). I am really quite impressed by the } } INCA, Oxford Instruments are, at last, listening to the users and adding } user } } requested facilities. They have sorted out the file handling mess of the } ISIS } } and structured it well for an SEM user (not quite as well for a TEM user } but } } there are less of us). The software structure is quite intuitive and there } is } } a really impressive help menu and explanation of everything from the } physics } } of X-ray generation, how EM's work, how detectors work and how to analyze } } samples. } } } } Their detectors have always been good and the SATW (thin window detectors) } } still have a reasonable efficiency at low Z. B is possible but C is easy } and } } even the N peak is over 30% efficient (there is often a high absorption at } N). } } } } Another feature that is invaluable for TEM is the integral shutter that } will } } close when the count rate is too high. This protects the crystal, it } prevents } } it overloading and shutting down or worse the crystal efficiency may } change } } for a few minutes until it recovers fully. This may affects your } quantitative } } work. In TEM this is usually caused by hitting the grid bar and not really } a } } problem in SEM but I don't know what secondaries and ions you will have in } a } } variable pressure SEM. It could be useful for you, check with other high } } pressure SEM users. } } } } Regards, } } Ron } } } } Please note: Oxford Instruments have upgraded an ISIS to an INCA system in } my } } department, without charge, in return for access to the instrument for } } development projects and demonstrations for a fixed number of days. I } receive } } no benefit from this and the department has no benefit from Oxford } Instruments } } sales. I remain a thorn in the flesh of all our suppliers if I think they } } could improve their products or service. } } ron.doole-at-materials.oxford.ac.uk } } } } } } -------------------------------------------------------------------------- } ---- } } } } } } Hello, } } I am very familiar with the Oxford ISIS 300 series spectrometers. They are } } ok, and the new Inca system looks good too. However, I recently saw the } PGT } } spectrometer at Lehigh and it is very impressive. } } } } Steve } } Stephen_Skirius-at-bkitech.com } } } } } } } } -------------------------------------------------------------------------- } } } } } } I'm also in the market for an EDS system and have looked at EDAX, Noran, } } PGT and Oxford. } } } } I edited out PGT because in order to quantify you have to optimize the } } system for the type of sample by playing with fudge factors, which none of } } the other systems have to do (though one of them, I think Noran, lets you } } adjust a sensitivity factor if you want to, but they didn't do it on my } } samples that were tested against known microprobe results and the answers } } were fine). I also eliminated Oxford, though it has a terrific user } } interface (maybe at the expense of functionality), because they } } consistently IDed my aluminum peaks as Br or Tm (!); this made me wonder } } about all their algorithms. They claim it had to do with the takeoff angle } } on the particular SEM being used, but that shouldn't be a factor. } } } } I like both EDAX and Noran, though for different reasons. EDAX user } } interface is better than Noran's, though again, I think Noran possibly } } offers more routines (it's hard to keep track and see absolutely } everything } } a system has to offer in a demo day....).Noran can multitask - work on } } several programs while a spectral map is being collected, for instance } } (does EDAX? I have to check). But EDAX has a beam skirt reduction routine } } for low vac mode (though it's time consuming, so a bit cumbersome), and } } their peak modeling is right up front - but Noran can put theirs up front } } also if you want to have it accessible (yes) and I think Noran might be a } } little better engineered. } } } } As you can see I'm still in a quandary (ditto for the two contending SEMs, } } LEO and ESEM). Whatever I decide I'll still be very interested in the } } results of your posting - especially if other folks' info comes in within } } the next week or so it would help in my decision too. } } } } I hope my input helps a little. Good luck with your quest! } } } } Dee Breger } } micro-at-ldeo.columbia.edu } } } } } } -------------------------------------------------------------------------- } - } } } } } } I looked into Noran, EDAX, and PGT. Noran was quickly culled (less user } } friendly, less abilities, didn't work right during demo), but EDAX and PGT } } both seemed to have equivalent capabilities (PGT claimed a 'proprietary' } } signal amplifier/digitizer doohickey, but it was only in the placement.) } } For the long-time spectrum gathering (I forget the technical term), PGT } } makes many passes with short dwell times while EDAX dwells on each pixel } } much longer to collect data & does it in 1 pass. Kinda 6 of one, half } dozen } } of the other. What made us choose the EDAX Phoenix system was the fact } that } } the PGT software was UNIX-based (although hidden) while EDAX is PC-based. } } I've heard rumors that PGT is switching to PC-based; we purchased our } system } } in 1999. I've also heard that Noran has practically no service techs (but } } that may be an East coast thing.) We've been happy with the EDAX service, } } and I enjoyed their user school very much. By the way, our SEM is a } } variable pressure JEOL 5900, and it's integrated with the EDAX system. } } } } Hope I've been helpful, } } } } Jane L. LaGoy } } Development Engineer } } Bodycote IMT, Inc. } } 155 River Street } } Andover, MA 01810 } } 978-470-1620 } } jlagoy-at-bodycote-imt.com } } } } } } } } } } }
{HTML} {FONT FACE=arial,helvetica} Jean, {BR} {BR} I would caution biased opinions from installation engineer and sales {BR} representatives ie... James Fotinopoulos.... www.semguy.com... {BR} {BR} Hmmmmmmmmm? {BR} {BR} Food for thought {BR} {FONT SIZE=2} {BR} {BR} {BR} {BR} {BR} ----- Original Message ----- {BR} {/FONT} {FONT COLOR="#000000" SIZE=3 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {/FONT} {FONT COLOR="#000000" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} From: "jeanross" <jeanross-at-blue.weeg.uiowa.edu> {BR} To: "Microscopy Listserver" <Microscopy-at-sparc5.microscopy.com> {BR} Sent: Tuesday, April 10, 2001 10:22 AM {BR} Subject: EDS summary {BR} {BR} {BR} > ------------------------------------------------------------------------ {BR} > The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} > To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} > On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} > -----------------------------------------------------------------------. {BR} > {BR} > {BR} > I have put together a summary of responses I got from my inquiry about a {BR} week {BR} > or so ago about EDS systems. I really appreciate everyone's input. We {BR} > haven't made any decisions yet since we are still gathering information {BR} but {BR} > your responses will help. I've included the responses in their entirety {BR} so I {BR} > hope this helps others as well. {BR} > {BR} > Thanks again from everyone who contributed. {BR} > {BR} > Jean Ross {BR} > Central Microscopy Research Facility {BR} > University of Iowa {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > I have been using an IXRF EDS / Gresham Detector (with IXRF digital pulse {BR} > processor) system for almost 2 years. Initially, the IXRF was installed on {BR} > an ETEC. About 15 months ago the Etec was replaced with a Hitachi 3500, a {BR} > new detector (Gresham) was purchased, and the EDS was installed on the {BR} 3500. {BR} > Generally, I am quite satisfied. There are minor software bugs, but IXRF {BR} > has been reasonably good at fixing them when discovered. It has been my {BR} > experience that all the systems have bugs, perhaps some more than others. {BR} > Prior to the IXRF, I had a Kevex 8000/Delta. {BR} > {BR} > Low end noise and broad peaks were evident on first installation, but were {BR} > soon fixed by tweaking the detector preamp and pulse processor amp. {BR} > {BR} > I am still running their first software package, "Iridium". I have the {BR} > newest release, "EDS 2000", but lack of time has kept me from installing {BR} and {BR} > checking it out. {BR} > {BR} > I should mention that IXRF is a "virtual" company, with people spread out {BR} > between Texas, California, etc. This has not proven to be a problem. {BR} > {BR} > Woody White {BR} > McDermott Technology, Inc. {BR} > nwwhite-at-mcdermott.com {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We have had EDAX for about a decade and a half, and we are very pleased {BR} with {BR} > the product and the service we get. {BR} > Carol Heckman {BR} > heckman-at-bgnet.bgsu.edu {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > look at IXRF eds systems there web site is www.ixrfsystems.com, they are {BR} > very affordable and offer no nonsense performance that second to none. {BR} > {BR} > happy ixrf user, {BR} > {BR} > James Fotinopoulos {BR} > yzfrjim-at-ix.netcom.com {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > I would recommend you give consideration to Doug Connors at {BR} > TN Analyzer Service, Inc. of Dane, WI. Doug has rebuild and {BR} > upgraded detectors for me for the last 6 years. He is dependable {BR} > knowledgeable, and economical as well. {BR} > {BR} > Bob Roberts {BR} > EM Lab Services, Inc. {BR} > 2409 S. Rural Rd Suite C {BR} > Tempe, Arizona 85282 {BR} > (480) 967-3946 {BR} > bobrobs-at-earthlink.net {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We recently purchased a Noran Instruments Vantage DS1. We purchased this {BR} > based on some very impressive demonstrations of software that the {BR} salesperson {BR} > brought in. Unfortunately they are still working out the bugs in their {BR} > software. Everything they have is ported over from Unix, and literally {BR} runs {BR} > in a unix shell on an Microsoft Windows NT platform. This makes their {BR} > software fairly buggy. Their response time to fix major bugs and hang-ups {BR} in {BR} > the software has been very slow, and if given the opportunity to do it all {BR} > over again I'd probably look at Oxford Instruments. I would still rate {BR} the {BR} > quality of the equipment very high. Our detector performs at the {BR} specified {BR} > resolution, and is a good piece of equipment. Now if they could only get {BR} the {BR} > software end of it straight... {BR} > My vote: {BR} > 1) Oxford Instruments {BR} > 2) Noran Instruments {BR} > 3) Edax or some of the smaller players {BR} > The benefit with going with a larger company is support and upgrades. We {BR} have {BR} > a 10 year old WDX that we just purchased new software and interface for {BR} last {BR} > year. Our old EDS was given some trade in value by Noran. And we all {BR} know {BR} > how valuable a service contract can be... {BR} > {BR} > Get back to me if you have any more questions, {BR} > ~Jonathan {BR} > Jonathan Dunlap {BR} > Analytical Laboratory Manager {BR} > Osram Sylvania Inc. {BR} > 816 Lexington Avenue {BR} > Warren, PA 16365 {BR} > Ph: 814-726-6991 {BR} > Fax: 814-726-6942 {BR} > Jonathan.Dunlap-at-sylvania.com {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > {BR} > We have an Oxford Instruments Link ISIS Model 200 on our 2460N. We have {BR} been {BR} > happy with it, but I don't know about the direction that Oxford is {BR} heading. I {BR} > don't care for the feel of their new INCA software. Some might like it. It {BR} > also seems to be slow coming together. Some of the functions are still {BR} lacking {BR} > after 2 (or is it 3) years of seeing it at MSA. {BR} > {BR} > We have an IXRF system on our JEOL 840A. It was a good price ($30K) for an {BR} > upgrade to our Kevex several years ago. It does what we need. They keep at {BR} > work on the software and have it freely available on the web. I might have {BR} to {BR} > pay closer attention and stay away from the beta stuff. They are still {BR} working {BR} > on it. They also have a nice digital pulse processor which stills stand {BR} alone {BR} > for about $5k. {BR} > {BR} > I still feel funny about some contacts with EVEX. I can't say much about {BR} EDAX, {BR} > NORAN, or PGT. They should all have good stuff but it might be pricey. The {BR} > last we seriously looked at them was 6 years ago or so when we opted for {BR} the {BR} > Oxford. {BR} > {BR} > I was intrigued by the unit from Quartz PCI. I think it was called X-ray {BR} One, {BR} > or such. It was new at MSA 1-1/2 years ago but looked promising. {BR} > {BR} > Feel free to call if you want more details. {BR} > {BR} > Warren E Straszheim {BR} > wesaia-at-iastate.edu {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We purchased an EDAX Falcon system for our Hitachi S-3000N and I've been {BR} > pleased with it. It has better light element sensitivity than most which {BR} > was very important to me although I don't think that its mapping {BR} > capabilities are as good as PGT's, say. I don't have direct experience {BR} with {BR} > Noran although I did talk to them and their system seemed ok - but {BR} logistics {BR} > didn't favor Noran so I passed on them. EDAX does have good integration {BR} > with the Hitachi and the Quartz database. {BR} > {BR} > I'd be glad to respond more specifically if you'd like. {BR} > {BR} > Richard Shalvoy {BR} > Arch Chemicals {BR} > Cheshire, CT {BR} > RBShalvoy-at-archchemicals.com {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > I have an iXRF systems out of Texas using a Gresham detector. It works {BR} > well. Not the most cutting edge, but they are one of the "start ups". They {BR} > have been around for I guess 6-7 years. I have a digital pulse processor {BR} > and completely active control for x-ray maps and such. They are very price {BR} > competitive, but lack a dedicated technical support person. You talk to {BR} the {BR} > programmer or electronics expert, but no techs on the phone whenever you {BR} > have a software question. But, if you willing to wait a day for some {BR} > answers then they are worth it. I haven't run across the problem where I {BR} > thought, "if I just had a better system". If you want to integrated w/ WDS {BR} > than maybe Noran. Also, if you want to integrated w/ motorized stage {BR} > control, I don't think they off such a package, like the bigger companies. {BR} > {BR} > I have a Hitachi 450. I used to run a 2400 and 500 before I quit my day {BR} job {BR} > and went out on my own. I am very happy w/ Hitachi. {BR} > {BR} > Good Luck {BR} > {BR} > Fell free to call with any specifics. {BR} > {BR} > Their web page is www.ixrfsystems.com {BR} > {BR} > Ric {BR} > {BR} > SMARTech {BR} > 860-491-3299 {BR} > www.semguy.com {BR} > 19 Cornwall Drive {BR} > Goshen CT 06756 {BR} > smartech-at-javanet.com {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > {BR} > Hello, all: {BR} > {BR} > I use Oxford ISIS300 system on HITACHI S-3500N (with VP mode) for light {BR} > element analysis, mostly C, O, N, F, P, S, Si, Mg, as well as metal Co, {BR} Ni-P, {BR} > Pt, Cr, Fe, W, etc. This system works well. One useful function is the {BR} overlay {BR} > of 2 spectrums. I can easily subtract the blank from the sample spot and {BR} make {BR} > it easy to identify what is (are) in the sample. I am sure some other {BR} program {BR} > may have this kind of function, but I have not seen. {BR} > {BR} > Zhiyu Wang {BR} > zhiyuw-at-home.com {BR} > I would be interested in seeing the responses as I am going to try and get {BR} > funding next year for a replacement for our EDAX PV9100 on an Hitachi {BR} s-450. {BR} > {BR} > Dave {BR} > David.Patton-at-uwe.ac.uk {BR} > {BR} > -------------------------------------------------------------------------- {BR} --- {BR} > {BR} > {BR} > We have been running EDX on SEMs and TEMs for many years. We used to have {BR} a {BR} > range of systems from Kevex, PGT, Noran, Link, EDAX, however a few years {BR} ago {BR} > we decided that we ought to standardize on one common system. After {BR} evaluation {BR} > we bought three Oxford Instruments ISIS systems. Whenever we have upgraded {BR} or {BR} > bought new systems they have been Oxford Instruments ISIS or now INCA. {BR} > {BR} > I have been happy with the ISIS except for the file handling that was not {BR} > designed for a multi user facility such as ours (approx 120 EM users in {BR} total {BR} > roughly 25 to 30 swapping every year). I am really quite impressed by the {BR} > INCA, Oxford Instruments are, at last, listening to the users and adding {BR} user {BR} > requested facilities. They have sorted out the file handling mess of the {BR} ISIS {BR} > and structured it well for an SEM user (not quite as well for a TEM user {BR} but {BR} > there are less of us). The software structure is quite intuitive and there {BR} is {BR} > a really impressive help menu and explanation of everything from the {BR} physics {BR} > of X-ray generation, how EM's work, how detectors work and how to analyze {BR} > samples. {BR} > {BR} > Their detectors have always been good and the SATW (thin window detectors) {BR} > still have a reasonable efficiency at low Z. B is possible but C is easy {BR} and {BR} > even the N peak is over 30% efficient (there is often a high absorption at {BR} N). {BR} > {BR} > Another feature that is invaluable for TEM is the integral shutter that {BR} will {BR} > close when the count rate is too high. This protects the crystal, it {BR} prevents {BR} > it overloading and shutting down or worse the crystal efficiency may {BR} change {BR} > for a few minutes until it recovers fully. This may affects your {BR} quantitative {BR} > work. In TEM this is usually caused by hitting the grid bar and not really {BR} a {BR} > problem in SEM but I don't know what secondaries and ions you will have in {BR} a {BR} > variable pressure SEM. It could be useful for you, check with other high {BR} > pressure SEM users. {BR} > {BR} > Regards, {BR} > Ron {BR} > {BR} > Please note: Oxford Instruments have upgraded an ISIS to an INCA system in {BR} my {BR} > department, without charge, in return for access to the instrument for {BR} > development projects and demonstrations for a fixed number of days. I {BR} receive {BR} > no benefit from this and the department has no benefit from Oxford {BR} Instruments {BR} > sales. I remain a thorn in the flesh of all our suppliers if I think they {BR} > could improve their products or service. {BR} > ron.doole-at-materials.oxford.ac.uk {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} ---- {BR} > {BR} > {BR} > Hello, {BR} > I am very familiar with the Oxford ISIS 300 series spectrometers. They are {BR} > ok, and the new Inca system looks good too. However, I recently saw the {BR} PGT {BR} > spectrometer at Lehigh and it is very impressive. {BR} > {BR} > Steve {BR} > Stephen_Skirius-at-bkitech.com {BR} > {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} > {BR} > {BR} > I'm also in the market for an EDS system and have looked at EDAX, Noran, {BR} > PGT and Oxford. {BR} > {BR} > I edited out PGT because in order to quantify you have to optimize the {BR} > system for the type of sample by playing with fudge factors, which none of {BR} > the other systems have to do (though one of them, I think Noran, lets you {BR} > adjust a sensitivity factor if you want to, but they didn't do it on my {BR} > samples that were tested against known microprobe results and the answers {BR} > were fine). I also eliminated Oxford, though it has a terrific user {BR} > interface (maybe at the expense of functionality), because they {BR} > consistently IDed my aluminum peaks as Br or Tm (!); this made me wonder {BR} > about all their algorithms. They claim it had to do with the takeoff angle {BR} > on the particular SEM being used, but that shouldn't be a factor. {BR} > {BR} > I like both EDAX and Noran, though for different reasons. EDAX user {BR} > interface is better than Noran's, though again, I think Noran possibly {BR} > offers more routines (it's hard to keep track and see absolutely {BR} everything {BR} > a system has to offer in a demo day....).Noran can multitask - work on {BR} > several programs while a spectral map is being collected, for instance {BR} > (does EDAX? I have to check). But EDAX has a beam skirt reduction routine {BR} > for low vac mode (though it's time consuming, so a bit cumbersome), and {BR} > their peak modeling is right up front - but Noran can put theirs up front {BR} > also if you want to have it accessible (yes) and I think Noran might be a {BR} > little better engineered. {BR} > {BR} > As you can see I'm still in a quandary (ditto for the two contending SEMs, {BR} > LEO and ESEM). Whatever I decide I'll still be very interested in the {BR} > results of your posting - especially if other folks' info comes in within {BR} > the next week or so it would help in my decision too. {BR} > {BR} > I hope my input helps a little. Good luck with your quest! {BR} > {BR} > Dee Breger {BR} > micro-at-ldeo.columbia.edu {BR} > {BR} > {BR} > -------------------------------------------------------------------------- {BR} - {BR} > {BR} > {BR} > I looked into Noran, EDAX, and PGT. Noran was quickly culled (less user {BR} > friendly, less abilities, didn't work right during demo), but EDAX and PGT {BR} > both seemed to have equivalent capabilities (PGT claimed a 'proprietary' {BR} > signal amplifier/digitizer doohickey, but it was only in the placement.) {BR} > For the long-time spectrum gathering (I forget the technical term), PGT {BR} > makes many passes with short dwell times while EDAX dwells on each pixel {BR} > much longer to collect data & does it in 1 pass. Kinda 6 of one, half {BR} dozen {BR} > of the other. What made us choose the EDAX Phoenix system was the fact {BR} that {BR} > the PGT software was UNIX-based (although hidden) while EDAX is PC-based. {BR} > I've heard rumors that PGT is switching to PC-based; we purchased our {BR} system {BR} > in 1999. I've also heard that Noran has practically no service techs (but {BR} > that may be an East coast thing.) We've been happy with the EDAX service, {BR} > and I enjoyed their user school very much. By the way, our SEM is a {BR} > variable pressure JEOL 5900, and it's integrated with the EDAX system. {BR} > {BR} > Hope I've been helpful, {BR} > {BR} > Jane L. LaGoy {BR} > Development Engineer {BR} > Bodycote IMT, Inc. {BR} > 155 River Street {BR} > Andover, MA 01810 {BR} > 978-470-1620 {BR} > jlagoy-at-bodycote-imt.com {BR} > {BR} > {BR} > {BR} > {BR} {BR} {/XMP} {/FONT} {FONT COLOR="#0f0f0f" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0"} {BR} {/FONT} {/HTML}
Return-Path: {ptarq-at-evex.com} Received: from rly-xb03.mx.aol.com (rly-xb03.mail.aol.com [172.20.105.104]) by air-xb02.mail.aol.com (v77_r1.36) with ESMTP; Tue, 10 Apr 2001 17:56:34 -0500 Received: from mxr01.nyc01.dsl.net (mxr01.nyc01.dsl.net [216.175.203.53]) by rly-xb03.mx.aol.com (v77_r1.36) with ESMTP; Tue, 10 Apr 2001 17:56:05 -0400 Received: from pb133nt (64-51-105-68.client.dsl.net [64.51.105.68]) by mxr01.nyc01.dsl.net (Postfix) with SMTP id 1F49C34538 for {lhconsulting-at-aol.com} ; Tue, 10 Apr 2001 17:52:57 -0400 (EDT) Message-ID: {002b01c0c208$95dfe860$6601a8c0-at-evex.com} Reply-To: "Evex Mail" {ptarq-at-evex.com} } From: "Evex Mail" {ptarq-at-evex.com} To: {lhconsulting-at-aol.com}
I guess it was a slip: In a sputter coater you can do rotary coating but shadowing is possible in evaporators only. For carbon coating the sputter coater would need an attachment for carbon string evaporation, which may be used for SEM and analyses, but not in TEM. Disclaimer: ProSciTech is the EMITECH instrument distributor in Australasia. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, April 11, 2001 5:18 AM, Gang Ning [SMTP:gning-at-mcw.edu] wrote: } } } Hi All: } } I want to buy a new/used sputter coater which enables to do rotary } shadowing as well as carbon coating. Any suggestions/input are } appreciated. } } Greg Ning } } EM Facility } Medical College of Wisconsin }
A colleague and I each recently bought Microtek scanners to scan TEM negatives. I have the Artixscan 1100 and he has the Model 8700 which has similar characteristics (actually higher resolution -1200dpi), 3.9 dmax at 42 bits color (14 grayscale), and the glassless film carrier setup. The 8700 has USB and Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a SCSI interface. You might want to check out the specs of the lower cost model 8700 on the microtekusa website if your computer can handle USB or Firewire. Both scanners have performed up to our expectations, which I would characterize as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier for standard size TEM film but you can easily make a serviceable one from stiff paper or light cardboard.
How much scanner resolution should you buy? The answer depends on how you intend to use it. Most applications do not require capturing the full resolution of the negative. From a practical viewpoint, the scanner resolution just determines how many times you can magnify the negative image to produce the final print size. For example, to get a publication-size print at 300 dpi, an image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to spending more for higher scanning resolution is to take photos at higher magnification. One exception is with lattice images from the TEM, which (depending on the lattice fringe spacing on the negative) might require higher scan resolutions to avoid getting a moire effect. (Of course, not everyone agrees. My colleague prefers to always scan at the maximum resolution).
What does a Dmax of 3.9 mean to you? To me it means a very dark negative. D is the log of the transmitted to incident intensity ratio. I wonder if users ever actually verify the manufacturer's specs with a calibrated density target. A Dmax of 3.9 can be useful for scanning TEM diffraction patterns that might have high contrast, but TEM micrograph negatives of metals and ceramics generally don't have that much contrast and biological thin section photos tend to have rather weak contrast. If your negatives are simply dark, use shorter photo exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 bits rather than 8) is highly recommended if you intend to enhance or adjust images, but that's another story.
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
---------- From: Tom Phillips Sent: Tuesday, April 10, 2001 11:22 AM To: Alwyn Eades Cc: Microscopy-at-sparc5.microscopy.com Subject: Re: Scanners
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I too am about to buy and I would make a couple of comments on your evaluation. First, let me remind everyone that the Dynamic range is a log scale so small numerical differences are significant.
I also think the Nikon Coolscan 8000 looks great but it only takes a 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone to a smaller film size or is Nikon using a non-Japanese EM as their standard? seems odd but I don't see how the Nikon would be very useful. You say a {2000 line scanner would be useful 9 out of 10 times but want the 2000+ lines for the occasional high res scan. I would argue that the size of the negative was the more important variable to be worried about. The Nikon couldn't handle 4x5 LM negatives or transparencies from autoradiography of Westerns/Northerns, etc.
My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. more details at www.microtek.com. This is my leading candidate. It was 4 negative carriers and I await word whether one could be modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have my scientific instrumentation shop guys fabricate a holder. It comes with a glass 8 x 10 glass carrier for odd size negs but I want to avoid Newton rings and want a glassless carrier.
I would appreciate comments on the following argument (I think I have this correctly figured out but am not sure since so many out there seem to want to have a higher resolution scanner). I have a Fuji Pictrography 3000 printer with a 400 dpi output that is as good as any other widely available printer in the academic world. If you figure the maximum published image size is about 8 inches, that would mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% depending on the orientation of the negative and still be taking full advantage of the printer resolution. In reality, most EM publication prints are smaller than 8" wide so one could crop even more and still not need more than 1000 dpi. A resolution } 1000 dpi would be useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x 4 negative would be 192 MB. That is pretty big for doing morphometry on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB and that is much more manageable. Perhaps the difference is in the type of EM we are doing. I am working with biological specimens doing standard thin section type stuff. are you doing some Material Sci application that demands more?
I will be interested in Alwyn (and any others) reply since I hope to buy one soon!
} . } } } There was a thread recently on scanners for TEM film. I have looked up } all the models mentioned, on the web and called agents for prices - and } produced a comparative table, given below. } } I do not guarantee that the figures are accurate but they are my best } interpretation of the data given. } } In the light of experience and Nestor's comments, I would suggest that } 2000 dpi is a minimum for TEM negatives. You may be able to get away } with less nine times out of ten, but there will be occasions when you } need more. } I would exclude the Minolta and all the Epsons from consideration } (despite the incredibly low prices of some of the Epsons) because of the } low pixel density. } } Among the rest the Nikon has the best pixel density and the best optical } density (another critical parameter for TEM negatives). The price is } very competitive too. The Nikon web site does not give a time for } scanning a negative. On the face of it the Nikon would be a best buy - } get a separate, inexpensive flatbed scanner for the other work. } } These comments are all my own opinions based on manufacturers' data. } Since we are considering purchase any comments to the contrary would be } most welcome. } } } } Code Maker Model Type } } } } A Agfa DuoScan T2500 Flatbed } -Transparency included } } B Epson 1640 several versions Flatbed } -Transparency option } 1680 several versions } } C 1600 several versions Flatbed } -Transparency included } } D Imacon Flextight Precision II Drum -for } film and large format } } E Minolta Dimage ScanMulti II Film } } F Nikon Super Coolscan 8000ED Film } } G Polaroid 45 Ultra Film } } H Umax Powerlook 3000 Flatbed } -Transparency included } } } } } } Code dpi OD Time Price } Opinion } at 6 x 9 cm } } } A 2500 x2500 3.4 3 min } $4,500 Fair } } B 1600 x 3200 3.6 } $300-$3000 Poor } } $800-$1400 Poor } } C 1600 x 3200 3.3 } $650-$1160 Not suitable } } D 2240 x2240** 3.9/4.1 N/A above } $10k Good: low pixel density } } E 1128 x 1128 3.6 } Not suitable } } F 4000 x 4000 4.2 N/A } $2,695 V. Good } } G 2500 x 2500 3.8 5 min } $7,495 Good but pricey } } H 3048 x 3048 3.6 3 min } $6,499 Good } } } -- } .......... } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvania 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Noise is only part of the problem, or solution, when considering a quantitative approach to scanning in negative or positive images. Since noise in electronic detection systems is largely dependent on temperature, that must also be brought into consideration. But the characteristics of the detector can be even more important. An array type device, such as a CCD, can have characteristics that vary from pixel to pixel as sensitivity, dynamic range and noise susceptibility. Your desire for a Consumer's Report on scanners is probably most appropriate as these various effects are difficult, if not impossible, to measure in a lab. Even if possible, these measurements may not be extendable to similar machines that aren't individually tested. Best to try to find a consensus on visual traits.
On Tuesday, April 10, 2001 7:50 AM, Sinkler, Wharton [SMTP:WSinkler-at-uop.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Laurie and Gary, } } Would noise be a good criterion? Say, for a perfectly evenly darkened film } (if such a thing existed, or at least even on a scale { { collected pixel } size) - what is the value of the noise (standard deviation of pixel value) } as a function of film darkness (density)? } } This would presumably improve with the time of collection. Thus how "good" } your scanner is depends on how you run it or whether it lets you take a } slower scan or to average multiple scans. With the exception of drum } scanners these devices all use CCD arrays. So what is probably most of } interest is the signal to noise ratio as a function of illumination } intensity, with everything known about CCD's going into determining this. } The maximum density the scanner can handle is just the point at which the } noise swamps the signal. } } There must be some good literature out there on the sources of noise, } optimizing collection (scan) time etc. One article which might be a } starting point is: } } G. H. Campbell, W. E. King and D. Cohen "Analysis of Experimental Error in } High Resolution Electron Micrographs", Microscopy and Microanalysis vol. 3 } (1997) p. 451. } } This is not very detailed, and treats only the total random noise, i.e. } grouping noise arising in collecting the image with that arising from the } scanner. } } Now, finding a good "Consumer Report" test with hard numbers on commercial } models is likely to be a lot harder! } } Wharton } } } -----Original Message----- } } From: Gary Gaugler [SMTP:gary-at-gaugler.com] } } Sent: Monday, April 09, 2001 10:37 PM } } To: L. D. Marks } } Cc: MSA listserver } } Subject: Re: Scanners: quantitative accuracy } } } } -------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } How do you define "quantitative digitization?" i.e., what } } variables are you dealing with in this respect? What are } } the "absolute terms?" } } } } Anyone else have some ideas about this topic? } } } } gg } } } } } } At 10:10 AM 4/9/2001, you wrote: } } } } } I have been listening to the thread on scanners. Has anyone done } } } tests of how accurate they are in absolute terms for quantitative } } } digitization? } } } } } } ------------------------------------------------------- } } } Laurence Marks } } } Department of Materials Science and Engineering & } } } Center for Transportation Nanotechnology } } } Northwestern University } } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } } mailto:ldm-at-risc4.numis.nwu.edu } } } http://www.numis.nwu.edu http://www.ctn.northwestern.edu } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Question: My student accidently got immersion oil on the 40x Leica Plan Acromat. I took a chance and used a fine artist's brush and xylene to clean it recalling (I may be wrong) that lens adhesives are soluble in alcohols not xylene, toluene and similar. Well, the lens didn't fall out. Too bad because I need an excuse to upgrade! If (or when) this happens again, what would you recommend for cleaning?
Does anyone out there have any experience microtoming bone that has NOT been decalcified? Is it even possible? I'm hoping to be able to have an answer for the admin people--prior to simply trying it out and possibly damaging my diamond knife.
Don Moravits Senior Technician Southwest Research Institute 6220 Culebra Road San Antonio, Texas 78238
I am embedding feather barbs which are mostly spongey dry collagen with keratin and small air pockets.
I am attempting Jan Dycks method-
1. 0.25 M NaOH 30 mins 2. formic acid / absolute alcohol 2:3 2 hrs ( to fill air pockets with solution) 3. 15% epon / propylene oxide 3 days 4. standard graduated increase of epon / dehydrant
Any other suggestions? Possibly from other similar material such as plant material.
Thanks
Tim Quinn Kansas University Museum of Natural History Lawrence, KS 785-864-4556
I would love to take advantage of the Firewire option but my information is that the 8700 has a Dmax of 3.4 vs the 3.9 for the 1100. That is a significant difference. Do EM negatives of biological thin sections reach that? I think so. I do a lot of EM immunocytochemistry and have to look for gold (intensely black) against a very dark tissue component so I am hoping the higher Dmax improves my results. I frequently scan negatives on a Umax 1100 (Dmax 3.4??) and can't differentiate the gold from the background although by eye I can discriminate them when the negative is placed on a light box. Changing my exposure would give me an unuseable image for the rest of the tissue. Maybe this is an extreme case but I suspect that lots of "dark organelles" (e.g., lysosomes, nuclei) have fine structure that get lost in the scanning with a low Dmax scanner. Tom
} A colleague and I each recently bought Microtek scanners to scan TEM } negatives. } I have the Artixscan 1100 and he has the Model 8700 which has similar } characteristics (actually higher resolution -1200dpi), 3.9 dmax at } 42 bits color } (14 grayscale), and the glassless film carrier setup. The 8700 has USB and } Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a } SCSI interface. You might want to check out the specs of the lower cost model } 8700 on the microtekusa website if your computer can handle USB or Firewire. } Both scanners have performed up to our expectations, which I would } characterize } as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier } for standard } size TEM film but you can easily make a serviceable one from stiff paper or } light cardboard. } } How much scanner resolution should you buy? The answer depends on how you } intend to use it. Most applications do not require capturing the full } resolution of the negative. From a practical viewpoint, the scanner } resolution } just determines how many times you can magnify the negative image to } produce the } final print size. For example, to get a publication-size print at 300 dpi, an } image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to } spending more for higher scanning resolution is to take photos at higher } magnification. One exception is with lattice images from the TEM, which } (depending on the lattice fringe spacing on the negative) might require higher } scan resolutions to avoid getting a moire effect. (Of course, not everyone } agrees. My colleague prefers to always scan at the maximum resolution). } } What does a Dmax of 3.9 mean to you? To me it means a very dark } negative. D is } the log of the transmitted to incident intensity ratio. I wonder if } users ever } actually verify the manufacturer's specs with a calibrated density target. A } Dmax of 3.9 can be useful for scanning TEM diffraction patterns that } might have } high contrast, but TEM micrograph negatives of metals and ceramics generally } don't have that much contrast and biological thin section photos tend to have } rather weak contrast. If your negatives are simply dark, use shorter photo } exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 } bits rather than 8) is highly recommended if you intend to enhance or adjust } images, but that's another story. } } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } } } ---------- } From: Tom Phillips } Sent: Tuesday, April 10, 2001 11:22 AM } To: Alwyn Eades } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: Scanners } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } I too am about to buy and I would make a couple of comments on your } evaluation. First, let me remind everyone that the Dynamic range is } a log scale so small numerical differences are significant. } } I also think the Nikon Coolscan 8000 looks great but it only takes a } 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM } negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone } to a smaller film size or is Nikon using a non-Japanese EM as their } standard? seems odd but I don't see how the Nikon would be very } useful. You say a {2000 line scanner would be useful 9 out of 10 } times but want the 2000+ lines for the occasional high res scan. I } would argue that the size of the negative was the more important } variable to be worried about. The Nikon couldn't handle 4x5 LM } negatives or transparencies from autoradiography of } Westerns/Northerns, etc. } } My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about } $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. } more details at www.microtek.com. This is my leading candidate. It } was 4 negative carriers and I await word whether one could be } modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have } my scientific instrumentation shop guys fabricate a holder. It comes } with a glass 8 x 10 glass carrier for odd size negs but I want to } avoid Newton rings and want a glassless carrier. } } I would appreciate comments on the following argument (I think I have } this correctly figured out but am not sure since so many out there } seem to want to have a higher resolution scanner). I have a Fuji } Pictrography 3000 printer with a 400 dpi output that is as good as } any other widely available printer in the academic world. If you } figure the maximum published image size is about 8 inches, that would } mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my } negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% } depending on the orientation of the negative and still be taking full } advantage of the printer resolution. In reality, most EM publication } prints are smaller than 8" wide so one could crop even more and still } not need more than 1000 dpi. A resolution } 1000 dpi would be } useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x } 4 negative would be 192 MB. That is pretty big for doing morphometry } on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB } and that is much more manageable. Perhaps the difference is in the } type of EM we are doing. I am working with biological specimens } doing standard thin section type stuff. are you doing some Material } Sci application that demands more? } } } I will be interested in Alwyn (and any others) reply since I hope to } buy one soon! } } } } . } } } } } } There was a thread recently on scanners for TEM film. I } have looked up } } all the models mentioned, on the web and called agents for } prices - and } } produced a comparative table, given below. } } } } I do not guarantee that the figures are accurate but they are my best } } interpretation of the data given. } } } } In the light of experience and Nestor's comments, I would suggest that } } 2000 dpi is a minimum for TEM negatives. You may be able to get away } } with less nine times out of ten, but there will be occasions when you } } need more. } } I would exclude the Minolta and all the Epsons from consideration } } (despite the incredibly low prices of some of the Epsons) because of } the } } low pixel density. } } } } Among the rest the Nikon has the best pixel density and the best } optical } } density (another critical parameter for TEM negatives). The price is } } very competitive too. The Nikon web site does not give a time for } } scanning a negative. On the face of it the Nikon would be a best buy } - } } get a separate, inexpensive flatbed scanner for the other work. } } } } These comments are all my own opinions based on manufacturers' data. } } Since we are considering purchase any comments to the } contrary would be } } most welcome. } } } } } } } } Code Maker Model Type } } } } } } } } A Agfa DuoScan T2500 Flatbed } } -Transparency included } } } } B Epson 1640 several versions Flatbed } } -Transparency option } } 1680 several versions } } } } C 1600 several versions Flatbed } } -Transparency included } } } } D Imacon Flextight Precision II Drum -for } } film and large format } } } } E Minolta Dimage ScanMulti II Film } } } } F Nikon Super Coolscan 8000ED Film } } } } G Polaroid 45 Ultra Film } } } } H Umax Powerlook 3000 Flatbed } } -Transparency included } } } } } } } } } } } } Code dpi OD Time Price } } Opinion } } at 6 x 9 cm } } } } } } A 2500 x2500 3.4 3 min } } $4,500 Fair } } } } B 1600 x 3200 3.6 } } $300-$3000 Poor } } } } $800-$1400 Poor } } } } C 1600 x 3200 3.3 } } $650-$1160 Not suitable } } } } D 2240 x2240** 3.9/4.1 N/A above } } $10k Good: low pixel density } } } } E 1128 x 1128 3.6 } } Not suitable } } } } F 4000 x 4000 4.2 N/A } } $2,695 V. Good } } } } G 2500 x 2500 3.8 5 min } } $7,495 Good but pricey } } } } H 3048 x 3048 3.6 3 min } } $6,499 Good } } } } } } -- } } .......... } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvania 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Does anyone know where I can find a multiple staining device for TEM grids using UA and Reynolds lead citrate that will result in CLEAN grids. Thank you Connie rosscac-at-okstate.edu
} Name: Kade } Organization: RMIT Melbourne Australia } Education: Graduate College } Location: Melbourne, Victoria, Australia } } Question: Hi, I have just purchased a microscope, a good biological } one. I need a microscope kit but nobody sells them around here. } Anyway already I have glass slides and covers. I have read some on } microscopy and I need an adhesive, resin I think its called to } prepare slides? is this true? } Also some ink to stain specimens. What are the names of all these } chemicals so I can buy them all seperatly since no one sells them all } together.
Kade -
You need a book on "microtechnique" plus a general idea of the types of specimens that you want to look at, BEFORE you start ordering things. Even tho you've "read some on microscopy " you could start with this book:
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with gool illustrations) of commonly encountered organisms. Adult. RECOMMENDED
You will find a lot of help on the amateur microscopy website http://www.microscopy-uk.org.uk
Both of these listings are from the Project MICRO bibliography (URL below).
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Your information is correct and mine is not. The Dmax of the 8700 is 3.4.
Larry
---------- From: Tom Phillips Sent: Wednesday, April 11, 2001 8:22 AM To: Microscopy-at-sparc5.microscopy.com Cc: jae5-at-lehigh.edu; Thomas, Larry (PNNL) Subject: RE: Scanners
I would love to take advantage of the Firewire option but my information is that the 8700 has a Dmax of 3.4 vs the 3.9 for the 1100. That is a significant difference. Do EM negatives of biological thin sections reach that? I think so. I do a lot of EM immunocytochemistry and have to look for gold (intensely black) against a very dark tissue component so I am hoping the higher Dmax improves my results. I frequently scan negatives on a Umax 1100 (Dmax 3.4??) and can't differentiate the gold from the background although by eye I can discriminate them when the negative is placed on a light box. Changing my exposure would give me an unuseable image for the rest of the tissue. Maybe this is an extreme case but I suspect that lots of "dark organelles" (e.g., lysosomes, nuclei) have fine structure that get lost in the scanning with a low Dmax scanner. Tom
} A colleague and I each recently bought Microtek scanners to scan TEM } negatives. } I have the Artixscan 1100 and he has the Model 8700 which has similar } characteristics (actually higher resolution -1200dpi), 3.9 dmax at } 42 bits color } (14 grayscale), and the glassless film carrier setup. The 8700 has USB and } Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a } SCSI interface. You might want to check out the specs of the lower cost model } 8700 on the microtekusa website if your computer can handle USB or Firewire. } Both scanners have performed up to our expectations, which I would } characterize } as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier } for standard } size TEM film but you can easily make a serviceable one from stiff paper or } light cardboard. } } How much scanner resolution should you buy? The answer depends on how you } intend to use it. Most applications do not require capturing the full } resolution of the negative. From a practical viewpoint, the scanner } resolution } just determines how many times you can magnify the negative image to } produce the } final print size. For example, to get a publication-size print at 300 dpi, an } image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to } spending more for higher scanning resolution is to take photos at higher } magnification. One exception is with lattice images from the TEM, which } (depending on the lattice fringe spacing on the negative) might require higher } scan resolutions to avoid getting a moire effect. (Of course, not everyone } agrees. My colleague prefers to always scan at the maximum resolution). } } What does a Dmax of 3.9 mean to you? To me it means a very dark } negative. D is } the log of the transmitted to incident intensity ratio. I wonder if } users ever } actually verify the manufacturer's specs with a calibrated density target. A } Dmax of 3.9 can be useful for scanning TEM diffraction patterns that } might have } high contrast, but TEM micrograph negatives of metals and ceramics generally } don't have that much contrast and biological thin section photos tend to have } rather weak contrast. If your negatives are simply dark, use shorter photo } exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 } bits rather than 8) is highly recommended if you intend to enhance or adjust } images, but that's another story. } } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto:Larry.Thomas-at-pnl.gov } } } } ---------- } From: Tom Phillips } Sent: Tuesday, April 10, 2001 11:22 AM } To: Alwyn Eades } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: Scanners } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } I too am about to buy and I would make a couple of comments on your } evaluation. First, let me remind everyone that the Dynamic range is } a log scale so small numerical differences are significant. } } I also think the Nikon Coolscan 8000 looks great but it only takes a } 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM } negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone } to a smaller film size or is Nikon using a non-Japanese EM as their } standard? seems odd but I don't see how the Nikon would be very } useful. You say a {2000 line scanner would be useful 9 out of 10 } times but want the 2000+ lines for the occasional high res scan. I } would argue that the size of the negative was the more important } variable to be worried about. The Nikon couldn't handle 4x5 LM } negatives or transparencies from autoradiography of } Westerns/Northerns, etc. } } My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about } $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. } more details at www.microtek.com. This is my leading candidate. It } was 4 negative carriers and I await word whether one could be } modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have } my scientific instrumentation shop guys fabricate a holder. It comes } with a glass 8 x 10 glass carrier for odd size negs but I want to } avoid Newton rings and want a glassless carrier. } } I would appreciate comments on the following argument (I think I have } this correctly figured out but am not sure since so many out there } seem to want to have a higher resolution scanner). I have a Fuji } Pictrography 3000 printer with a 400 dpi output that is as good as } any other widely available printer in the academic world. If you } figure the maximum published image size is about 8 inches, that would } mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my } negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% } depending on the orientation of the negative and still be taking full } advantage of the printer resolution. In reality, most EM publication } prints are smaller than 8" wide so one could crop even more and still } not need more than 1000 dpi. A resolution } 1000 dpi would be } useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x } 4 negative would be 192 MB. That is pretty big for doing morphometry } on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB } and that is much more manageable. Perhaps the difference is in the } type of EM we are doing. I am working with biological specimens } doing standard thin section type stuff. are you doing some Material } Sci application that demands more? } } } I will be interested in Alwyn (and any others) reply since I hope to } buy one soon! } } } } . } } } } } } There was a thread recently on scanners for TEM film. I } have looked up } } all the models mentioned, on the web and called agents for } prices - and } } produced a comparative table, given below. } } } } I do not guarantee that the figures are accurate but they are my best } } interpretation of the data given. } } } } In the light of experience and Nestor's comments, I would suggest that } } 2000 dpi is a minimum for TEM negatives. You may be able to get away } } with less nine times out of ten, but there will be occasions when you } } need more. } } I would exclude the Minolta and all the Epsons from consideration } } (despite the incredibly low prices of some of the Epsons) because of } the } } low pixel density. } } } } Among the rest the Nikon has the best pixel density and the best } optical } } density (another critical parameter for TEM negatives). The price is } } very competitive too. The Nikon web site does not give a time for } } scanning a negative. On the face of it the Nikon would be a best buy } - } } get a separate, inexpensive flatbed scanner for the other work. } } } } These comments are all my own opinions based on manufacturers' data. } } Since we are considering purchase any comments to the } contrary would be } } most welcome. } } } } } } } } Code Maker Model Type } } } } } } } } A Agfa DuoScan T2500 Flatbed } } -Transparency included } } } } B Epson 1640 several versions Flatbed } } -Transparency option } } 1680 several versions } } } } C 1600 several versions Flatbed } } -Transparency included } } } } D Imacon Flextight Precision II Drum -for } } film and large format } } } } E Minolta Dimage ScanMulti II Film } } } } F Nikon Super Coolscan 8000ED Film } } } } G Polaroid 45 Ultra Film } } } } H Umax Powerlook 3000 Flatbed } } -Transparency included } } } } } } } } } } } } Code dpi OD Time Price } } Opinion } } at 6 x 9 cm } } } } } } A 2500 x2500 3.4 3 min } } $4,500 Fair } } } } B 1600 x 3200 3.6 } } $300-$3000 Poor } } } } $800-$1400 Poor } } } } C 1600 x 3200 3.3 } } $650-$1160 Not suitable } } } } D 2240 x2240** 3.9/4.1 N/A above } } $10k Good: low pixel density } } } } E 1128 x 1128 3.6 } } Not suitable } } } } F 4000 x 4000 4.2 N/A } } $2,695 V. Good } } } } G 2500 x 2500 3.8 5 min } } $7,495 Good but pricey } } } } H 3048 x 3048 3.6 3 min } } $6,499 Good } } } } } } -- } } .......... } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvania 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear all, We are having a great deal of difficulty getting hold of our normal film "Tritiated Hyper film" for use with tritiated ligands. Does any one out there know of a local supplier (U.K.)
We use 70 % isopropanol to clean emersion oil off of our lenses.
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Wed, 11 Apr 2001 mckaylodge-at-aol.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Email: mckaylodge-at-aol.com } Name: Robert Lodge } } Organization: McKay Lodge Home School } } Education: 9-12th Grade High School } } Location: Oberlin, OH 44074 } } Question: My student accidently got immersion oil on the 40x Leica } Plan Acromat. I took a chance and used a fine artist's brush and } xylene to clean it recalling (I may be wrong) that lens adhesives are } soluble in alcohols not xylene, toluene and similar. Well, the lens } didn't fall out. Too bad because I need an excuse to upgrade! If (or } when) this happens again, what would you recommend for cleaning? } } Bob Lodge } } --------------------------------------------------------------------------- }
Has anyone had experience in determining the concentration of particles in a solution by counting on EM grids in a similar way to using a haemocytometer? Is it possible to count the particles in a defined number of grid squares then calculate back to the area of the grid and the amount of solution which was applied and allowed to dry down?
Richard Gardiner Department of Plant Sciences University of Western Ontario
Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is discontinued. Any suggestions for an alternative? We have been using Kodabrome RC II for a long time with a Mohr processor. How about Polycontrast RC? Or is this the beginning of the end for old fashioned darkrooms?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
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One of the big advantages of computer printing of TEM images is the ability to adjust the contrast of the final image. However, to prevent generating a negative that has completely overexposed areas which are difficult to salvage by any means-such as one may get with metal or ceramic specimens-an alternative film developer may help. There are two bath "split" developers which lower contrast to manageable levels, in effect chemically "dodging" a developing negative. Fixing is carried out normally. I'm told biological specimens don't usually need such treatment. I have no financial interest in the following company, but I am pleased with the results obtained with their developer called Diafine. It is made by Acufine Inc., 5441 North Kedzie Ave. Chicago, Il., 60625.
Bernard Kestel E-mail: {kestel-at-anl.gov} Materials Science Division Argonne National Laboratory Argonne, Il., 60439
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id TAA26206 for dist-Microscopy; Wed, 11 Apr 2001 19:12:49 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id TAA26197 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 11 Apr 2001 19:12:19 -0500 (CDT) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id TAA26190 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Apr 2001 19:12:08 -0500 (CDT) Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {p05001900b6faa35d2003-at-[206.69.208.21]}
Below is the result of your feedback form. It was submitted by (jgh7f0-at-mizzou.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 11, 2001 at 14:32:21 ---------------------------------------------------------------------------
Email: jgh7f0-at-mizzou.edu Name: john harris
Organization: university of missouri-columbia
Education: Undergraduate College
Location: Columbia, Missouri
Question: Dear Sir or Madam; i am looking for an image of S. Agalactiae attached to an epithelial cell or some other gram + cocci.
Kodabrome is for sure an old product. But a good one. I quit using it long ago in favor of Ilford paper. Ilford makes RC paper--but my work has only been output to archival fiber paper. But I suspect that the RC quality is still up to par.
I use plain matte for normal prints--and archival matte fiber for fine art prints (nudes, etc.). I use a Kreonite processor for the print papers. All b/w neg media is hand processed in separate roll holders (Nikor). My normal format is 6x4.5cm and 6x7cm. The same recipes would apply to other formats.
I did some 4x5" work in prior years and do the same mechanics today.
Try some Ilford paper.
gary g.
http://photoweb.net
At 03:25 PM 4/11/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Wow...this is really deja vu. I used Diafine and Acufine way many years ago. At that time, the issue was push processing. Today, the zone system prevails. Using the zone system does not require special chemicals. It is a matter of how the neg is rated and how it is processed.
Look up some of Ansel Adams' works (The Negative). The main idea is to adjust your neg's EV for total tonal range such that it can be printed with less than bone crushing effort.
All of my fine art prints are shot and printed using this process.
gary g.
http://photoweb.net
At 04:41 PM 4/11/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used Illiford RC paper for a long time and I liked it better than anything else I have ever used. It may be just be personal preference but it seemed to have brighter higlights and blacker blacks than I could get with other RC papers. Just be sure and get Illford filters to go with it. One other nice thing is it has a matt finsh as well as glossy if you don't want glossy prints. The matt will scan great. It doesn't have texture problems like most pearl or simi-glossy papers have. The matt finish displays a lot better than glossy finsh IMHO.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} From: "Gary Gaugler" {gary-at-gaugler.com}
} Kodabrome is for sure an old product. But a good one. } I quit using it long ago in favor of Ilford paper. Ilford } makes RC paper--but my work has only been output to archival } fiber paper. But I suspect that the RC quality is still up to par. } } I use plain matte for normal prints--and archival matte fiber } for fine art prints (nudes, etc.). I use a Kreonite processor } for the print papers. All b/w neg media is hand processed } in separate roll holders (Nikor). My normal format is 6x4.5cm } and 6x7cm. The same recipes would apply to other formats. } } I did some 4x5" work in prior years and do the same } mechanics today. } } Try some Ilford paper. } } gary g. } } http://photoweb.net } } } } } Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is } } discontinued. Any suggestions for an alternative? We have been using } } Kodabrome RC II for a long time with a Mohr processor. How about } } Polycontrast RC? Or is this the beginning of the end for old fashioned } } darkrooms? } } } } Jonathan Krupp
} I am using Polymax II RC. For draft pictures it's good enough in my point } of view. In compare with Ilford Multigrade III RC, Polymax gives better } contrast (filter #5 on Ilford is equal to #3 on Polymax in my hands).
Sergey
} At 03:25 PM 4/11/2001, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Usualy to reduce contrast you under expose the film. Then develop it to the disired density using a developer that generates low contrast. One way to reduce contrast is to dilute you developer by a factor of 2, 4 or more with water. It extends the developing time a good deal but it reduces the contrast. You might also look at low contrast developers that work with the film you are using.
A few question on rec.photo.darkroom will get you more information than you can handle and some of it will actually work.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} } } } } } One of the big advantages of computer printing of TEM images is the } } ability to adjust the contrast of the final image. However, to prevent } } generating a negative that has completely overexposed areas which are } } difficult to salvage by any means-such as one may get with metal or ceramic } } specimens-an alternative film developer may help. } } There are two bath "split" developers which lower contrast to } } manageable levels, in effect chemically "dodging" a developing negative. } } Fixing is } } carried out normally. I'm told biological specimens don't usually need } } such treatment. } } I have no financial interest in the following company, but I am } } pleased with the results obtained with their developer called Diafine. It is } } made by Acufine Inc., 5441 North Kedzie Ave. Chicago, Il., 60625. } } } } Bernard Kestel E-mail: {kestel-at-anl.gov} } } Materials Science Division } } Argonne National Laboratory } } Argonne, Il., 60439 } }
-----Original Message----- } From: christine [mailto:ac.richardson2-at-btinternet.com] Sent: Wednesday, April 11, 2001 1:59 PM To: microscopy-at-sparc5.microscopy.com
Dear all, We are having a great deal of difficulty getting hold of our normal film "Tritiated Hyper film" for use with tritiated ligands. Does any one out there know of a local supplier (U.K.)
Back in the pre PC (chemical) days, toluene was a recommended solvent for cleaning lenses of immersion oil. Now, we at National Steel, wipe off excess oil with lens tissue and then clean the lens with Kodak lens cleaner solution.
Best regards,
Sam Purdy National Steel Tech Center Trenton MI
} ---------- } From: mckaylodge-at-aol.com } Sent: Wednesday, April 11, 2001 9:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist:Help Cleaning Lenses } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Email: mckaylodge-at-aol.com } Name: Robert Lodge } } Organization: McKay Lodge Home School } } Education: 9-12th Grade High School } } Location: Oberlin, OH 44074 } } Question: My student accidently got immersion oil on the 40x Leica } Plan Acromat. I took a chance and used a fine artist's brush and } xylene to clean it recalling (I may be wrong) that lens adhesives are } soluble in alcohols not xylene, toluene and similar. Well, the lens } didn't fall out. Too bad because I need an excuse to upgrade! If (or } when) this happens again, what would you recommend for cleaning? } } Bob Lodge } } -------------------------------------------------------------------------- } - }
Kodabrome has not been discontinued across the board. However, certain size/surface/grade combinations have been. Kodabrome II RC F5 is available in 250sheet 8x10 packages only. We stock them but a dealer should be able to order them. As far as alternatives.... Agfa offers Brovira Speed RC, a developer incorporated paper in grades 2-5 and available in 100 sheet 8x10 packs. Brovira Speed is a cold tone paper. Agfa also offers a Multicontrast RC product. From Kodak, Polycontrast or Polymax are variable contrast papers. Polycontrast is developer incorporated, as Kodabrome is, so it will process in developers as well as many activators. Polymax requires the use of a developer and will not process in activators. Ilford offers Multigrade IV Deluxe which is not developer incorporated. All of these papers use filters to control contrast, although a #5 filter with any of them is not the same as a grade 5 Kodabrome. Both Polymax and Multigrade IV have a slightly wider contrast range than Polycontrast. For those who prefer a cooler tone print, Ilford also has a new paper, Multigrade Cooltone, a non developer incorporated paper with a cooler image tone and a cool white base tint.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
} Just tried to order some Kodabrome II RC paper, grade F5. Vendor } says it is discontinued. Any suggestions for an alternative? We have } been using Kodabrome RC II for a long time with a Mohr processor. How } about Polycontrast RC? Or is this the beginning of the end for old } fashioned darkrooms? } } Jonathan Krupp
The paper is still listed on the Kodak Web sight, http://www.kodak.com/cluster/global/en/professional/support/techPubs/f33/old/f33.shtml . The web sight does list Kodabromide paper as discontinued but lists Kodabrome II RC as its replacement and does not list the latter as discontinued. I was curious, so, I called the 800 technical hotline (800) 242-2424 (option 02 gives you an operator) and asked. They said that it is still available but not in as many sizes as it use to be.
I've had venders tell me a product is discontinued when it is only discontinued from their stocking process, not by the manufacture. In this case it may be that only the size you wanted was discontinued. You may want to call the vendor back or check with another vender.
I have no connection with Kodak but wanted to see if the predictions I'd heard about, as you put it, " the beginning of the end for old fashioned darkrooms," was starting. It would seem not quite yet in this case. Though I'm hearing that in 2 or 3 years it will, somewhat sad I think.
Jim Roberts
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } {jmkrupp-at-cats.ucsc.edu} 04/11/01 03:25PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi:
Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is discontinued. Any suggestions for an alternative? We have been using Kodabrome RC II for a long time with a Mohr processor. How about Polycontrast RC? Or is this the beginning of the end for old fashioned darkrooms?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Yes, Thermanox cover slips survive treatment with HMDS. We occationally process cell monlayers grown on them for SEM.
Regards,
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Thu, 12 Apr 2001, Randall, Kevin J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need to process some thermanox coverslips for SEM. Does anyone know } whether thermanox survives HMDS? } } Cheers } } Kevin }
Hi All: Thank you all for sharing your experiences about working with SiC. Your suggestions have helped me come up with a plan of attack. Regards, Mike Coviello UT Arlington
We are a Kodak distributor and we checked with Kodak this morning and Kodabrome II RC paper, grade F5 is still available. If you still interested you may contact me off-line.
John Arnott
Jon Krupp wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi: } } Just tried to order some Kodabrome II RC paper, grade F5. Vendor says it is } discontinued. Any suggestions for an alternative? We have been using } Kodabrome RC II for a long time with a Mohr processor. How about } Polycontrast RC? Or is this the beginning of the end for old fashioned } darkrooms? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
We are about to install a Kimball ES-423E (extended life) LaB6 cathode in a Hitachi H7100 TEM and were wondering if anyone had some starting points for the voltage settings for filament heating. This would save us a lot of time, if so.
Thank you.
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
Here is an odd way that I thought up to clean oil from an objective without using solvents that may attack lens cement.
Remove the lens from the scope. I usually like to view the oil contamination using a stereoscopic microscope. Wipe excess oil away - I like using Ross Optical Paper. Using a cotton applicator wrapped in Ross optical paper, apply a small amount of Dawn dishwashing detergent. Gently work the surface to emulsify the oil into the detergent using the optical papered applicator. You may have to add a small amount of water. Do not allow fluids to go much beyond the lens area!
At this point, hold the lens vertical with the back focal plane pointing up. Apply a small amount of deionized water on the final lens element from the side of the opjective to create a hanging drop. I usually use a wash bottle or 10cc syringe. The surface tension of water will create a small drop around the optical surface. Start a stream of DI water flowing through the small drop to wash away the oil/water suspension. After about a minute, stop the water flow while allowing a small drop of water to stay on lens. At this time, blow the water off using C02 or some other clean compressed gas. This will eliminate streaks. Upon inspection, if you see contamination, repeat the process and your problem should be solved.
It's weird but it works.
Robert
Robert Fitton Teaching Associate/Director of Laboratories Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 563-387-1559 FAX 563-387-1080
Enjoy a visit to our website: http://www.luther.edu/~biodept/
Dear Richard, The most common use of this is for asbestos fibre load determination. I did it once. You count the number of fibres in a specified number of grids openings, then calculate back to the original collected volume or vacuumed area. It should work similarly for any recognizable particle. I could look up the method, if you like. At 06:08 PM 4/11/01 -0400, you wrote: } } Has anyone had experience in determining the concentration of particles } in a solution by counting on EM grids in a similar way to using a } haemocytometer? Is it possible to count the particles in a defined } number of grid squares then calculate back to the area of the grid and } the amount of solution which was applied and allowed to dry down? } } Richard Gardiner
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Below is the result of your feedback form. It was submitted by (dngeorge-at-wam.umd.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 12, 2001 at 16:04:35 ---------------------------------------------------------------------------
Email: dngeorge-at-wam.umd.edu Name: Damali Martin
Organization: University of Maryland
Education: Graduate College
Location: College Park, MD
Question: I have embedded and sectioned some salivary glands in epon and have placed the sections on glass slides. However, when I counterstain, a high percentage of sections float off and are lost. How can this problem be prevented?
Also, several of my sections have wrinkles in them. Is there any trick to getting rid of them?
Anyone got any suggestions as to the cause of (and remedy for) the pronounced image shift that occurs when I turn the Y stigmator control on my JEOL 840A?
It also has the usual stigmatic effect.
The X control shifts the image only very little.
The coils seem to check out OK.
There's something quite poetic about working on this particular part of the instrument today of all days.
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Stigmators operate as two opposing coils. It may be that one of the coils for the Y stigmator is not active while the opposing Y coil is. Perhaps one of the coils has shorted? The resistance of these coils is very low, making a simple check rather difficult, not to mention that they are often series connected within the column making physical access difficult. But if you can measure the current drawn by each, you may find a significant difference if one is shorted.
If so, the only remedy is replacement of the coils.
On Friday, April 13, 2001 1:12 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } } Hi } } Anyone got any suggestions as to the cause of (and remedy for) the } pronounced image shift that occurs when I turn the Y stigmator } control on my JEOL 840A? } } It also has the usual stigmatic effect. } } The X control shifts the image only very little. } } The coils seem to check out OK. } } There's something quite poetic about working on this particular part } of the instrument today of all days. } } thanks } } rtch } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
About 25 years ago, when I was taking the embryology course at the Marine Biology Laboratory at Woods Hole, we were taught the following methods for cleaning oil from lenses by Robert Allen, a well known microscopist:
Remove the lens from the microscope and invert it. Remove most of the excess oil from the objective with lens paper without touching the objective surface itself. Gently lay a strip of lens paper over the objective. Since the lens is recessed the lens paper does not touch the glass directly. Then place a drop or two of ether on the lens paper just next to where it contacts the lens, and draw the lens paper over the objective surface. The ether vapors swirl around under the lens paper and dissolve the oil while the lens paper absorbs the mixture and carries it away, without actually touching the glass surface. You might need to repeat a few times to remove the oil.
It works with all but the most stubborn deposits. The upside is that it completely avoids touching the glass surface, and the ether fumes are in contact with the element such a short time that it miakes it less likely you will dissolve the glues that hold the lens elements together. The downside is that it requires ether. I haven't tried it with other solvents. -- Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
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Accountabilities % of time
70 Collect data from production activities to track and improve quality. Develop collection systems to optimize this process. We have a new Hitachi S3000N with an Oxford INCA EDS system and an Oxford/Gatan CL system with cryostage. Routine analysis will include EBIC, EDS, and CL.
20 Interface with Manufacturing Engineers, Application engineers, Detail Engineers, Product Managers, Technicians, Maintenance, Tool Room, Field Service, and other stakeholders to achieve assigned project goals. Recommend design changes to improve manufacturability and quality.
10 Coordinate project work activity, including testing, machining, and purchasing, as needed to ensure meeting project schedules. Design and build fixtures, prototypes, and samples. Provide training on equipment and techniques to assemblers.
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Learn about ADC - The Broadband Company at www.adc.com
I usually see image shift as a result of stigmation in both of our scopes (JEOL 840A and Hitachi 2460N). I rather accepted that it just went with the territory. If it is a sign of a problem with the scope or its alignment, I too would be interested in hearing how to eliminate it.
Warren
At 06:12 PM 4/13/2001 +0000, you wrote:
} Hi } } Anyone got any suggestions as to the cause of (and remedy for) the } pronounced image shift that occurs when I turn the Y stigmator } control on my JEOL 840A? } } It also has the usual stigmatic effect. } } The X control shifts the image only very little. } } The coils seem to check out OK. } } There's something quite poetic about working on this particular part } of the instrument today of all days. } } thanks } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The 840 has some pots (potentiometers) on one of the circuit boards that adjust out the image shift while adjusting the stigmators. They probably balance a bias, or an offset, in the circuit for the stigmation coils. If you have the manual with the schematics, they may be identified. I may be able to find it in ours, if you don't have it.
Normally, X or Y stig adjustment will cause image shifting. More shifting at higher mag. Since stigmation is done with coils by the scan coils, changing current in the stig coils appears to the beam as a change in scan coil current. Hence, the image shifts. Later day systems account for this by feeding some of the stig signal to the scan coils as feedback. It will also accept additional feedback from magnification. The operation is to adjust scan coil current and condenser current as stig is adjusted so that the net result is minimal image shift with varying stig input.
Since one stig channel out of two is not working right, there are likely one of two or three things which could go wrong. First, and worst, the stig coil is open or shorted. Somewhat unlikely I think. Second, one part of the stig coil drive circuit has failed. Third, some part of the feedback system has failed. Since you say that it does stig, but causes image shift, then the stig circuit itself and the coil is OK. You can verify this.
Since the stig coil drive systems are identical (ususally), you should be able to trace readings from the good channel and compare them to the bad channel. This should show right away where the problem is. The stig, beam alignment and image shift circuits are usually all the same. If so in your system, they will give plenty of data points for checking. The coils (one for X, one for Y) are typically driven by push-pull power transistors which are high current buffers on the output of a small op amp. The coils are in the negative feedback loop of the op amp. One lead of a coil would connect to the output of the buffer transistors (large ones) while the other end connects to the inverting input of the op amp and is returned to ground through a low value resistor. Measuring the voltage across this resistor will tell how much current is flowing through the coil (I=E/R).
Stig effect is lower at lower mags. So there would be less automatic compensation for stig vs. mag. Try a low mag setting and measure the voltage on each side of the stig coils (two leads each) and the voltage on the low value resistors. Have the stig controls at 12 O'clock each. If these readings match, odds are that the coils are for sure OK. Then the problem ought to be narrowed to the stig balance circuit. Look for this circuit and compare the two stig signal feedback loops for differences. It could be something as simple as a bad op amp in the path from the stig circuit to the beam alignment coil drivers. Since stig has little effect at low mag, but huge effect at high mag, the usual control path for stig compensaton versus magnification is to electronically change the beam position by sending a stig sourced voltage to the scan coil driver circuit.
If you don't have schematics for the system, that is of course a major problem. In this case, perhaps someone who has your same model has encountered this problem before and knows of the failure mechanism and cause.
gary g.
At 11:12 AM 4/13/2001, you wrote:
} Hi } } Anyone got any suggestions as to the cause of (and remedy for) the } pronounced image shift that occurs when I turn the Y stigmator } control on my JEOL 840A? } } It also has the usual stigmatic effect. } } The X control shifts the image only very little. } } The coils seem to check out OK. } } There's something quite poetic about working on this particular part } of the instrument today of all days. } } thanks } } rtch } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Stigmator shift is almost always down to the balance of the stigmator coils. Stigmators have 8 coils in four sets of two. If an opposing pair are not correctly balanced the stronger coil will cause the image to move away from that coil.
Some instruments have the balancing potentiometers easily accessible others hide them away in the electronics.
The pots will be called Xx, Xy, and Yx, Yy I do not know where they are on a JEOL 840 but if you look inside you may be lucky?
To adjust - 1. Place an easily recognizable feature in the center of the screen 2. Turn the X stigmator fully in one direction and re center with the Xx or Xy control whichever fits. 3. Turn in the opposite direction and use the other compensation control to center 4. Repeat with the Y stigmator and Yx and Yy controls.
I have just completed the promised "Monitoring & Maintaining Electron Microscope Performance" interactive CD, this area is covered in the instrument tuning section so its pretty fresh in my mind. Need to see even more for yourself then we have the course of the same name running in Sydney early October this year?
Kindest regards and good luck
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I have a Hitachi SEM model S-570 (1985). I am interested of getting a X-ray analyzer for this microscope. My preference would be to buy a used analyzer with an ultrathin window for light elements analysis.
If you know someone who is interested in selling it's analyzer, please let me know...
Thank you
Christian Normand Tekna Systems Plasma (819) 820-2204
I hesitated to make any response regarding this subject. But--here goes anyway. Someone may find it useful.
I think that the issue is not reduction of contrast but rather extension of tonal range of the negative. That is what the zone system is all about. I think that it is a fact that if the silver is not exposed, there is not going to be any information gleaned from that unexposed or too underexposed area. The approach I use for fine art images is to overexpose and under-develop. It takes a LOT of work to develop a complete system of ISO rating, development and printing to achieve marketable prints. But it certainly can be done. The goal is to minimize the time spent in the darkroom doing printing. Ideally, the neg should print without any effort on grade 3 paper. I only use Ilford archival matte fiber paper, so extensions to RC papers may or may not directly apply. I also do not use variable contrast paper. But the principles are extensible I would think.
Being all-digital with my SEM, I use realtime histogram feedback to adjust gain and dynamic range. For TEM, I have no direct experience. But I might suggest that the same photo techniques for fine art neg work may apply to TEM negs. If you think that this might be true, read on. Otherwise, nevermind.
A neg with bad dynamic range (huge extremes of EV or low contrast) is not going to produce a very good scan. This is the realm of drum scanners with 4.0-6.0D. Pixel resolution is subordinate to D range in this case. If one is going to print a neg on paper, that is one aspect of the problem. If the neg is to be output to a magazine or printed page, that is another aspect. So the crux of the matter is what the actual intended end use of the neg is to be? If it is a nice print, OK. If it is a magazine or litho output, these are 133lpi. Doing scans at 4000dpi only to print them in a magazine at 133lpi is rather silly. One would probably be better off just printing the neg and scanning at 300dpi.
But the 300 dpi is the effective dpi for the size of the final image....not the original neg. So, if the neg is say 3" x 4" and is to be printed at that same size, a 300 dpi scanner should do the job (ignoring tonal range for the time being). If the neg is to be printed at twice its physical size, then the scanner has to have twice the resolution (600 dpi). And it goes on from there. Thus, for most image output methods on a printed page, 600-1200 optical dpi should be plenty of resolution. If you agree with this discussion at this point, then the resolution factor should no longer be an issue. What remains is D range.
(Note that commercial images are created at high resolution to accommodate manipulation and merging with other images. This way, image quality can gradually be reduced without affecting the final result.)
But the negative is the key item in all of this discussion. A bad neg is not likely to do anyone much good. So the challenge is to get a good neg at the get go. Here we go, back to the zone system. The idea is to use the neg as a non-linear image capture media for data which may span a wide dynamic range. And in so doing, be able to output (print or scan) this information with fidelity and minimal effort. This drives D value.
An unexposed, developed neg will have a transmission of 100%. It is actually slightly lower due to absorption by the medium, emulsion and residual anti-dispersion coating. But at 100% transmission, the neg would have an opacity of 100%input/100%output or 1.0. Since D=log1/T, the D value for the unexposed neg is log 1/1=0.0. This then is Dmin. The area with the most exposure (darkest region) will pass the least amount of transmitted light. If, for example, this region passes 1% of the transmitted light, this is a Tval=100%/1%=100. And D=log (1/100)=2.0. This then is Dmax. The D range of the negative is Dmax-Dmin=2.0-0.0=2.0. So a scanner with a D value of } 2.0 would capture the tonal range of this neg. The tonal range of this neg is 100 tonal variations. Prints generally have D values between 1.7 and 2.0. So the same scanner would handle the neg and a typical print.
Let's say that the darkest area of the neg transmits 0.1% of the transmitted source light. This then yields Tval=100/0.1 =1000. And D=log 1000 = 3.0. Thus, this neg has 1,000 tonal variations in it from clear to black. If the neg has double the number of tonal variations (2,000) that would mean that the opacity is 2,000 and D=log 2000 = 3.3. At 4000 tonal range, D=3.6. Thus, each 0.3D equates to doubling the opacity range or halving or doubling the transmission value. Thus, if a scanner has a D rating of 3.0 (1000 tonal ranges) and the neg passes a corresponding 0.1% of the transmitted light, and another scanner has a D rating of 3.3 (2000 tonal ranges) and the neg passes 0.05% of the transmitted light--can you really tell one from the other?
The eye is more responsive to subtle changes in tonal variations in the white region of an image than in the dark ones. Thus, it is important to retain as much variability and rendition in the darkest areas of the negs (the highlights) but to do so without sacrificing detail in the shadows (clearer areas of the neg). This is done using the zone system and expansion and contraction of tonal range.
The response of the neg emulsion's exposure to light is not linear over the range of clear to full black. Its response curve is somewhat like a flattened S. The low exposure region is called the toe, which consists of the film base + fog density (Dmin). Moving up the curve is the straight line section (not exactly a straight line), and then to the shoulder. At this point, increasing amounts of light do not have corresponding quantitative amounts of change in density. Further exposure reaches saturation, or Dmax. At this point, no increase in exposure will change the density of the neg.
The ultimate goal is to maximize the straight line section and move Dmax as high as possible, without detrimentally affecting Dmin. I use contraction to accomplish this. The approach is to overexpose and underdevelop. With a neg, the rule of thumb is to expose for the shadows. If there is insufficient exposure in the shadow areas, there will not be any detail rendered.
Using Ilford FP4+ film, I rate it at ISO 80 (mfg=125). Development is done using stock developer diluted 1:1. It is used one time and discarded. Never replenish or try to refresh the developer. Development time will vary, depending on the tonal range of the scene. Here are some extreme examples of how this system works: (if you are offended by nude images, use the still life links. These following links are to fine art nudes and still life which were done using the zone system previously described)
[nudes] A. This shot illustrates a typical impossible shot. Inside the room, the exposure was about EV 3, the outside was dense fog with a starch white picket fence at EV 11. A scene like this with eight stops of variation would typically be shot to either render detail in the highlights (fog and fence) or in the inside room's details (shadows). To render shadow detail and retain highlight detail, the zone system was used as described. The image via this link is as-scanned on a UMAX Powerlook III of a 6x6cm negative using the transparency adapter. You can see the detail in the paint on the wall and can see the fence in the fog. http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_2.html
B. This shot shows great shadow detail despite the outside light creating a hot spot on the model's head. And the window on the left was rendered, despite the high level of light. http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_6.html
C. This shot shows a rather low contrast scene. There are no remarkable highlights. There is much material in shadow. Overexposure and ensuring at least two zones of exposure for the shadows and then overdeveloping N+1 achieved a perfectly printable neg. Again, this neg is shown as-scanned. http://www.photoweb.net/pw_gal_nude/pw_gal_nude_6/g_3.html
[Still life] A. high side lighting. http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_5.html
B. Low shadow lighting, highlights present. http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_6.html
C. Even lighting. There is actually more detail revealed than is showed in this web pix. http://www.photoweb.net/pw_gal_still/pw_gal_still_3/g_6.html
Non-web images of these negs are of course much better than those on-line. But I hope these illustrate the points of the zone system. I do think it can apply to TEM negs. The other point is to keep in mind the relationship of D values of scanners to what you are actually going to be scanning. If a scanner has sufficient resolution, and say a D rating of 3.2. Are you really going to be able to tell any difference using a scanner with a 3.4 or 3.5D at much higher cost? If you have a densitometer, check the D range of some of your negs. They are probably all less than 3.0. Maybe I'm wrong in this respect since I have little experience with TEM media. But the idea is go get the equipment you need for the job you need (the output destination and the use of the image) based on the actual media being scanned. Otherwise, there is a great opportunity to buy capability which will never be utilized.
Despite all the discussion of processing negs, as more digital capture and image processing products come out, things will change. There are rather simple ways to expand the contrast of an otherwise low contrast, poor neg. Likewise, there are ways to extract subtle detail from negs which have blown out highlights. More about this later.
gary g.
Reference: Adams, A. (1981). The negative. Boston: Little, Brown and Company. ISBN 0-8212-1131-5 (twelfth printing, 1992).
At 01:07 AM 4/12/2001, you wrote:
} Usualy to reduce contrast you under expose the film. Then develop it to } the disired density using a developer that generates low contrast. One way } to reduce contrast is to dilute you developer by a factor of 2, 4 or more } with water. It extends the developing time a good deal but it reduces the } contrast. You might also look at low contrast developers that work with } the film you are using. } } A few question on rec.photo.darkroom will get you more information than } you can handle and some of it will actually work. } } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger
Good job, Gary! Gary's description of dynamic range is the clearest and most easily understood I have seen (and I have been looking for a good one!). I think I already understood most of what he said before he said it but I was having a devil of a time trying to articulate the concept to one of my staff. My only followup question is in regards to his statement near the end where he suggests some test the density of a TEM negative with a densitometer to see if they really go much above 3.0. I would like to encourage any one who has or will be measuring this for a typical and even finicky biological thin section TEM image to please post the info on the Microscopy listserver. Thanks.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I hesitated to make any response regarding this subject. } But--here goes anyway. Someone may find it useful. } } I think that the issue is not reduction of contrast but rather extension } of tonal range of the negative. That is what the zone system is } all about. I think that it is a fact that if the silver is not exposed, } there is not going to be any information gleaned from that } unexposed or too underexposed area. The approach I use for fine } art images is to overexpose and under-develop. It takes a LOT } of work to develop a complete system of ISO rating, development } and printing to achieve marketable prints. But it certainly can } be done. The goal is to minimize the time spent in the darkroom } doing printing. Ideally, the neg should print without any effort } on grade 3 paper. I only use Ilford archival matte fiber paper, so } extensions to RC papers may or may not directly apply. I also } do not use variable contrast paper. But the principles are extensible } I would think. } } Being all-digital with my SEM, I use realtime histogram feedback } to adjust gain and dynamic range. For TEM, I have no direct } experience. But I might suggest that the same photo techniques } for fine art neg work may apply to TEM negs. If you think that this } might be true, read on. Otherwise, nevermind. } } } A neg with bad dynamic range (huge extremes of EV or low contrast) is not } going to produce a very good scan. This is the realm of drum } scanners with 4.0-6.0D. Pixel resolution is subordinate to } D range in this case. If one is going to print a neg on paper, } that is one aspect of the problem. If the neg is to be output } to a magazine or printed page, that is another aspect. So the } crux of the matter is what the actual intended end use of the } neg is to be? If it is a nice print, OK. If it is a magazine or litho } output, these are 133lpi. Doing scans at 4000dpi only to } print them in a magazine at 133lpi is rather silly. One would } probably be better off just printing the neg and scanning at } 300dpi. } } But the 300 dpi is the effective dpi for the size of } the final image....not the original neg. So, if the neg is say } 3" x 4" and is to be printed at that same size, a 300 dpi scanner } should do the job (ignoring tonal range for the time being). } If the neg is to be printed at twice its physical size, then the } scanner has to have twice the resolution (600 dpi). And it } goes on from there. Thus, for most image output methods } on a printed page, 600-1200 optical dpi should be plenty of } resolution. If you agree with this discussion at this point, } then the resolution factor should no longer be an issue. } What remains is D range. } } (Note that commercial images are created at high resolution } to accommodate manipulation and merging with other images. } This way, image quality can gradually be reduced without } affecting the final result.) } } But the negative is the key item in all of this discussion. A } bad neg is not likely to do anyone much good. So the challenge } is to get a good neg at the get go. Here we go, back to the } zone system. The idea is to use the neg as a non-linear } image capture media for data which may span a wide } dynamic range. And in so doing, be able to output (print } or scan) this information with fidelity and minimal effort. } This drives D value. } } An unexposed, developed neg will have a transmission of 100%. } It is actually slightly lower due to absorption by the medium, } emulsion and residual anti-dispersion coating. But at 100% } transmission, the neg would have an opacity of 100%input/100%output } or 1.0. Since D=log1/T, the D value for the unexposed neg } is log 1/1=0.0. This then is Dmin. The area with the most } exposure (darkest region) will pass the least amount of } transmitted light. If, for example, this region passes 1% of } the transmitted light, this is a Tval=100%/1%=100. } And D=log (1/100)=2.0. This then is Dmax. The D range } of the negative is Dmax-Dmin=2.0-0.0=2.0. So a scanner } with a D value of } 2.0 would capture the tonal range of this } neg. The tonal range of this neg is 100 tonal variations. } Prints generally have D values between 1.7 and 2.0. } So the same scanner would handle the neg and a typical } print. } } Let's say that the darkest area of the neg transmits 0.1% } of the transmitted source light. This then yields Tval=100/0.1 } =1000. And D=log 1000 = 3.0. Thus, this neg has 1,000 } tonal variations in it from clear to black. If the neg has double } the number of tonal variations (2,000) that would mean } that the opacity is 2,000 and D=log 2000 = 3.3. At 4000 } tonal range, D=3.6. Thus, each 0.3D equates to doubling } the opacity range or halving or doubling the transmission } value. Thus, if a scanner has a D rating of 3.0 (1000 tonal } ranges) and the neg passes a corresponding 0.1% of the } transmitted light, and another scanner has a D rating } of 3.3 (2000 tonal ranges) and the neg passes 0.05% of } the transmitted light--can you really tell one from the other? } } The eye is more responsive to subtle changes in tonal } variations in the white region of an image than in the } dark ones. Thus, it is important to retain as much variability } and rendition in the darkest areas of the negs (the highlights) } but to do so without sacrificing detail in the shadows (clearer } areas of the neg). This is done using the zone system } and expansion and contraction of tonal range. } } The response of the neg emulsion's exposure to light is not linear } over the range of clear to full black. Its response curve is somewhat } like a flattened S. The low exposure region is called the toe, which } consists of the film base + fog density (Dmin). Moving up the curve } is the straight line section (not exactly a straight line), and then to } the shoulder. At this point, increasing amounts of light do not } have corresponding quantitative amounts of change in density. } Further exposure reaches saturation, or Dmax. At this point, } no increase in exposure will change the density of the neg. } } The ultimate goal is to maximize the straight line section and } move Dmax as high as possible, without detrimentally affecting } Dmin. I use contraction to accomplish this. The approach is } to overexpose and underdevelop. With a neg, the rule of thumb } is to expose for the shadows. If there is insufficient exposure } in the shadow areas, there will not be any detail rendered. } } Using Ilford FP4+ film, I rate it at ISO 80 (mfg=125). Development } is done using stock developer diluted 1:1. It is used one time } and discarded. Never replenish or try to refresh the developer. } Development time will vary, depending on the tonal range of the } scene. Here are some extreme examples of how this system works: } (if you are offended by nude images, use the still life links. These } following links are to fine art nudes and still life which were done using the } zone system previously described) } } [nudes] } A. This shot illustrates a typical impossible shot. Inside the room, } the exposure was about EV 3, the outside was dense fog with } a starch white picket fence at EV 11. A scene like this with } eight stops of variation would typically be shot to either render } detail in the highlights (fog and fence) or in the inside room's } details (shadows). To render shadow detail and retain highlight } detail, the zone system was used as described. The image } via this link is as-scanned on a UMAX Powerlook III of a } 6x6cm negative using the transparency adapter. You can } see the detail in the paint on the wall and can see the fence in } the fog. } http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_2.html } } B. This shot shows great shadow detail despite the outside light } creating a hot spot on the model's head. And the window } on the left was rendered, despite the high level of light. } http://www.photoweb.net/pw_gal_nude/pw_gal_nude_1/g_6.html } } C. This shot shows a rather low contrast scene. There are no } remarkable highlights. There is much material in shadow. } Overexposure and ensuring at least two zones of exposure } for the shadows and then overdeveloping N+1 achieved } a perfectly printable neg. Again, this neg is shown as-scanned. } http://www.photoweb.net/pw_gal_nude/pw_gal_nude_6/g_3.html } } } } [Still life] } A. high side lighting. } http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_5.html } } B. Low shadow lighting, highlights present. } http://www.photoweb.net/pw_gal_still/pw_gal_still_4/g_6.html } } C. Even lighting. There is actually more detail revealed } than is showed in this web pix. } http://www.photoweb.net/pw_gal_still/pw_gal_still_3/g_6.html } } } } Non-web images of these negs are of course much better than } those on-line. But I hope these illustrate the points of the zone } system. I do think it can apply to TEM negs. The other point } is to keep in mind the relationship of D values of scanners to } what you are actually going to be scanning. If a scanner has } sufficient resolution, and say a D rating of 3.2. Are you } really going to be able to tell any difference using a scanner } with a 3.4 or 3.5D at much higher cost? If you have a } densitometer, check the D range of some of your negs. They } are probably all less than 3.0. Maybe I'm wrong in this } respect since I have little experience with TEM media. } But the idea is go get the equipment you need for the job } you need (the output destination and the use of the image) } based on the actual media being scanned. Otherwise, there } is a great opportunity to buy capability which will never be } utilized. } } Despite all the discussion of processing negs, as more } digital capture and image processing products come out, } things will change. There are rather simple ways to expand } the contrast of an otherwise low contrast, poor neg. Likewise, } there are ways to extract subtle detail from negs which have } blown out highlights. More about this later. } } gary g. } } } Reference: } Adams, A. (1981). The negative. Boston: Little, Brown and Company. } ISBN 0-8212-1131-5 (twelfth printing, 1992). } } } } At 01:07 AM 4/12/2001, you wrote: } } } Usualy to reduce contrast you under expose the film. Then develop it to } } the disired density using a developer that generates low contrast. One way } } to reduce contrast is to dilute you developer by a factor of 2, 4 or more } } with water. It extends the developing time a good deal but it reduces the } } contrast. You might also look at low contrast developers that work with } } the film you are using. } } } } A few question on rec.photo.darkroom will get you more information than } } you can handle and some of it will actually work. } } } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I tried sending this earlier, but seems to have not made it.
The 840 has some pots (potentiometers) on one of the circuit boards that are used to adjust out any image shift during stigmation adjustments. They probably adjust some bias voltages, or some offsets, in the stigmator circuits. They can be adjusted so there is no image shift.
If the pots can't eliminate the image shift, then a component has probably gone bad, rather than just drifted a bit. If you have the manual with the schematics, they should be identified in there. If you don't have the manual, I might be able to find them in ours. Let me know.
The 18th Annual NESM Woods Hole Symposium will be held May 11-12th, at the Marine Biological Lab at Woods Hole, MA.
This meeting is supported by members of the Connecticut Microscopy Society (CMS), the Metropolitan Microscopy Society (MMS), and the New York Society of Experimental Microscopists (NYSEM).
Pre-registration is encouraged and is a must if you plan to attend the Friday night dinner. Inquiries re: registration for this meeting should be directed to Mary McCann (617) 484-7865 or by email: mccanns-at-tiacc.net. Advance regis- tration including dinner on May 11th is $40.00 for NESM members, and $55.00 for non-members (this cost includes a one- year membership to NESM).
PLEASE NOTE: ADVANCE REGISTRATION MUST BE RECEIVED no later than MAY 4th!!! Registrations received after May 4th will NOT include dinner.
Friday, May 11th begins at Noon and consists of 2 sessions with an afternoon coffee break. Following the presentations, a cocktail hours and dinner will commence in the Swope Center.
Saturday, May 12th, NESM will present a symposium on Remote Access Microscopy. Afterwards, there will be commercial exhibits and posters on display in the Swope Center. Presentation of Poster and Photos-As-Art Awards and Door Prizes will follow. This year lunch at the Swope Center will be optional; those interested will pay $16.00. After lunch, two 45-minute tours of the Marine Resource Center will take place.
NESM welcomes new members to the Society! Please join us for this most enjoyable meeting.
Peggy Sherwood, Corresponding Secretary NESM -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
You have a really good point for discussion about actual available exposure time. Real world nudes and still lifes can be for several seconds exposure. What is it for a typical TEM neg? I don't know. I suspect the same drift problems occur in TEM as they do in SEM. So I too like to get the shot as quickly as possible yet with minimum noise. So it is a tradeoff of pixel density and pixel dwell time. Some original loss can be made up later using the computer. With a digital active scan and image capture system synchronized to 60 Hertz, a SEM shot at 3000x3200 pixels takes 96 seconds at 10uS dwell time. This works fine. But for a TEM which may not have sync to line frequency, I can see that image shift is a big problem. Are TEM cameras and scanning routinely synchronized to line frequency?
I used Diafine and Accufine back in photojournalism days to push process TriX to ISO1600 or beyond. The job was to shoot basketball and football games without strobe. The reason was to gain even lighting and high contrast shots. Since the pix were physically small, the higher grain was not a big issue.
Your Diafine approach to TEM negs sounds like a solid method. Indeed, the key is to get the shot and make a print with minimum amount of tweaking (burning, dodging, etc.). If one were to only have to make one print ever from one neg, it would not matter all that much. But if more than one print is or will be made, reproducibility is a tough issue with a less than perfect neg.
I see a similarity in your approach and what I was talking about. Your procedure limits the development time of the neg. But while it limits the time for dense areas, it limits the time for the whole neg too. The procedure I described ensures that as much light information is captured on the film, but extends the tonal range of the film by reducing the strength of the developer and the development time.
I already have de-rated the speed of the film in this procedure. It may be that doing this with TEM negs makes the resulting absolute speed too slow. What is the rated ISO speed of some TEM media in use today?
gary g.
At 01:56 PM 4/13/2001, you wrote: } Reply to: RE: Film Processing & dynamic range & scanners (longish) } I saved your extensive info about negs. I would not use Diafine for } regular scenic photos--too low in contrast. However, the manner in which } split developers achieve this is to LIMIT the amount of developer } available to "dense" negative areas. I only use it for contrasty TEM } negs. that I routinely print on #2 or #3 paper , whichever is more } pleasing. By having easily retrievable info in all of a negative, less } time should be needed by any final printing method. The developer has a } long tank life and works at room temperature. An extreme test of TEM negs } made at 4 f/stops equivalent underexposure still printed nicely on #3 } grade paper. } Of course I used a very long 12 mimutes in each half of the split } developer, but this can allow short exposure on specimens with a } "drifting" problem and save an expensive experiment. } As primarily an old "wet" printer, I enjoyed your thorough } explaination and have heard of the zone system and the great A. Adams. I } like to get good results with little effort, thats all. If you ever want } a copy of the Diafine Co.'s explaination of it's product, I can send you } one. Nearly all of my published photos in Ultramicroscopy were done with } this stuff, including a "lucky" cover photo on Ultra. 25 (1988) 351-354. } } Bernie Kestel E-mail: {kestel-at-anl.gov} } Materials Science Division } Argonne National Lab } 9700 So. Cass Ave., } Argonne Il., 60439 } Gary Gaugler wrote:
Has anyone had experience in determining the concentration of particles in a solution by counting on EM grids in a similar way to using a haemocytometer? Is it possible to count the particles in a defined number of grid squares then calculate back to the area of the grid and the amount of solution which was applied and allowed to dry down?
Dear Richard, I can forsee one difficulty. The particles may not be deposited uniformly due to a number of factors: 1) Evaporation of the applied drop of specimen will not be uniform, so the particles could be dragged in toward the grid center as the edges evaporate (or they could be preferrentially deposited toward the edges if they are hydrophobic). 2) The grid surface may not be flat, causing particles to deposit preferrentially in the centers of the grid squares--if these are lower--or near the grid bars. 3) The particles could be preferrentially deposited either on top of the grid bars or on the open areas, which could look like uniform deposition, but would give erronious quantitation. Of course, one could test this by depositing a known amount of suspension of a known concentration of the kind of particles to be measured, then seeing if the grid was covered uniformly and if the calculation gave the correct result. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I asked if xylene was the best solvent for cleaning immersion oil off a non-immersion objective. I recalled in my brief training that xylene was safe but alcohols dissolve the more common lens adhesives (I could be wrong and have this reversed). Here is the array of responses I received. The diversity may interest you. THANKS to all for your advice.
1 We regularly use xylene with a cotton tipped applicator to clean our lenses and remove immersion oil. This was recommended to us by the supplier of our lenses and microscopes for a number of reasons. The primary reason is that xylene effectively 'cuts' the oil, without damaging the lens. Acetone will affect the lens coating, and Isopropyl alcohol only rinses the oil without effectively removing it. Hope this helps
2 You are lucky that the lens elements did not dislodge. Use alcohol on lens paper. Never use xylene or toluene; these solvents can dissolve the cement used to seat the various lens elements.
3 I would recommend Sparkle Glass Cleaner. I only use solvents as a very last resort (unless you really, really do want to upgrade) The cements are soluble in most solvents. With intractable grime, I use a cotton tipped applicator moistened with either xylenes or toluene and shake all excess off. The tip must be moist, not wet or dry and make single passes until the grime come off. Only enough solvent on the applicator to dampen the surface, not enough to wet.
4 For day to day cleaning of lenses, including oil on the 40X. (I can't believe this is the first time! The graduate students and Post Doc's drag the40X through the oil at least twice a week. I have given them repeated instructions but they seem intractable) Invert an ocular and examine the surface for cleanliness. If it is clean, don't clean it. Dust off any loose debris. Check the lens again. Moisten a cotton tipped applicator in Sparkle and wipe the lens 1 swipe with a rolling action to present a new surface and lift off any grit. Discard. Take a dry applicator and remove the film. Check the lens with an inverted ocular to see if it is clean. Do no more than is necessary. There has been an ongoing rampage on the list about lens cleaning.... to solvent, not to solvent... lens tissue, not to lens tissue etc.. I can append you a couple, and a microscope maintenance handout if you would like. It will be a bit long about 15 pages (38K).
5 The care instructions for my immersion oil suggest cleaning with a soft cloth or lens tissue (no Kimwipes) moistened with ether/alcohol (7:3) or xylenes. My microscope manuals recommend removing finger prints using alcohol. Sounds like you're good to go.
6 We use 70 % isopropanol to clean emersion oil off of our lenses.
7 Cleaning with xylene is a little drastic. If the lens eventually gets xylene in behind the cement it could do some damage and you would have to send it in to Nikon for repair. I use Kodak lens cleaner and some lens cleaning tissues (not regular Kim wipes) to get the oil off, constantly checking them under a dissecting scope to make sure that it is all removed. It works well on some expensive lens that we have here.
8 We use Green Soap from the pharmacy. It works the best with ultra pure water.
9 Actually, you've got your solvents reversed; alcohol is usually safer. But safer still is diluted detergent "Joy" or similar, followed by water. Use alcohol only if that doesn't work. And fumes from xylene, toluene, etc. are best avoided.
10 Back in the pre PC (chemical) days, toluene was a recommended } solvent for cleaning lenses of immersion oil. Now, we at National Steel, } wipe off excess oil with lens tissue and then clean the lens with Kodak lens } cleaner solution.
Ritchie, What you are describing is a stigmator drive that is out of calibration. If you look on the schematic for the stigmator drive, you should see the X(or Y) control that goes to an op-amp that in turn feeds a voltage divider which in turn controls a different coil in the stigmator coil assembly. Each part of this voltage divider has its own trim pot for calibration purposes. Simply rock the X stigmator control back and forth while adjusting each pot for minimum image shift. Then, increase your mag & repeat. You should be able to get all of the shift completely out. Good luck!
Gary M. Easton, Pres. Scanners Corporation SEM/EDS/IMAGING Sales & Service
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, April 13, 2001 2:12 PM
Thanks very much to all those who responded to my post.
My hands didn't bleed at all.
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
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At 12:53 PM -0400 4/10/01, Ronald Anderson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am sending this message again because it was blocked by a list server filter - presumably because 'home' is part of your e-mail address.
Malcolm
-------- Original Message --------
Please reply directly to:
} From: {hmskaug-at-tartarus.uwa.edu.au} } } Subject: TEM } } } Hi, } } I am doing some preperation work for an assignment, and have a question. } } If a fresh unfixed piece of brain tissue (weighing approx. 1 gram) is } received in a diagnostic electron microscopy laboratory, how would I } proceed with the specimen? The brain is taken in surgery within the last 5 } min. } } If I was asked to carry out a rapid viral diagnosis on the tissue, what } precautions should I take, and how would I proceed? } I read that prions are able to survive fixation. What then? } } A respond is very much appreciated. } } Thank You ! } } Sincerely, } } Hege Skaug
Greg Erdos Assistant Director Biotechnology Program Ph. 352-392-1295 University of Florida Fax 352-846-0251 PO Box 118525 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
To all colleagues who supply standards for EDS work I need a standard sample of known thickness to check the accuracy of my TEM/EDS system during standardless quant. I need to include light elements (Z {11) as I have a Moxtek SUTW window on my detector. I guess an amorphous glass sample that contains both nitrogen and oxygen with with a very thin layer of carbon on it to prevent charging would do the trick. Can anybody out there supply one? Thanks.
Alan Fox
Professor Alan G. Fox BSc PhD CEng FIM Director, Center for Materials Science and Engineering Naval Postgraduate School Monterey California 93943 USA
This is an apology written in embarassment for some of my private comments about EDX systems that recently got broadcast to the list. I had too-quickly responded offline to a posting and was chagrined to find my comments passed along without the balanced context I would have put them in if I'd been more sensitive to the probability of dissemination. It's too late to rephrase them but I want to emphasize that my recent demos with Oxford, Noran, PGT and EDAX were all highly positive experiences and I came away believing that all are solid and supportive companies offering world class instrumentation. Any system will have many pros and a few cons; our final choice will rest simply on which has a bigger cluster of the former than the latter for our particular set of applications. The cons I had mentioned were subjectively-perceived blips within the significent strengths that each system offers. While cons can serve to pare down the choice, once it's made, as I've discovered through discussion with reference users representing all four companies, users across the board (at least all those I've spoken to) are happy with whichever system they purchased.
Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
There is a decent test sample out there for checking and monitoring your EDS system called the NiOx sample. One place that you can find it is at Ted Pella. Here is the web site for that product. It has further information on it. http://www.tedpella.com/calibrat_html/TEM7.htm
I am not sure what you mean by the accuracy of your EDS system. You will need to calibrate it and determine whether you are having problems with icing. You can do that with the NiOx sample. The literature that comes with the sample or that you can get from Ted Pella will tell you how to do this. It is also tells you how to monitor your system's performance over time.
If you want to check for N2, just get some Hexagonal BN, crush it between two glass slides, and collect it on a carbon coated grid. You will easily find thin particles.
You hit on a couple of problems with your request. You will have a difficult time ascertaining the thickness of any sample accurately, but with glass, you will only be limited to doing it with EELS or contamination spots. With glass, the composition can change under the beam as elements particularly alkali elements diffuse out of the irradiated area. The composition of glasses can change depending on the depth from the surface and so where you are can make a big difference. I do not know where you would get a glass with a N2 concentration. You can also cause the glass to soften in the beam in very thin areas if care is not taken. Higher accelerating voltages help here tremendously.
I have found that frequently I do not need to coat my glass samples in a 200 keV TEM. I did have to do it when I used a 100 keV machine. For cross section samples, I started using Si blanks as half of the sample and it seems to have eliminated heating and charging problems.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Alan Fox [mailto:fox-at-nps.navy.mil] } Sent: Monday, April 16, 2001 12:50 PM } To: MS listserver } Subject: Standard sample for EDS quant } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } ---------. } } } To all colleagues who supply standards for EDS work } I need a standard sample of known thickness to check the } accuracy of my TEM/EDS system during standardless quant. I need to } include light elements (Z {11) as I have a Moxtek SUTW window on my } detector. I guess an amorphous glass sample that contains } both nitrogen } and oxygen with with a very thin layer of carbon on it to prevent } charging would do the trick. Can anybody out there supply one? Thanks. } } Alan Fox } } } Professor Alan G. Fox BSc PhD CEng FIM } Director, Center for Materials Science and Engineering } Naval Postgraduate School } Monterey } California 93943 } USA } } Tel (831) 656 2142 (work) } (831) 657 9239 (home) } Fax (831) 656 2238 } } }
May I suggest that we consider some ground rules regarding postings of summaries of replies to questions? It happens on occasion that replies which are assumed to be made in private get posted publicly, sometimes to the embarrassment of those doing the replying. This is, I'm sure, never done with any bad intent and is usually harmless , but can nonetheless be a little disconcerting.
I have posted summaries myself without thinking of the possible consequences, so I'm not pointing any fingers. It's something that's easy to forget about until you get caught yourself.
I would suggest as starting points that: 1) questioners always make clear their intent to post a summary, 2) that the identities of the repliers be removed from summaries (this is often done anyway), and 3) that all who reply to a question indicate if they want their replies to be kept private.
Anyone else have any thoughts on this matter?
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
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I would need to prepare DNA for TEM using spreading/shadowing in Cytochrome C films. My pilot experiments using just plasmid DNA and the Lang & Mitani method (Biopolymers, 9, p.373, 1970) worked rather poorly and I would like to hear from people with some experience in this method. What are the critical points here? Purity of water/chemicals, Cyt C concentration, time? Any hints would be appreciated.
Do I understand you believe this is CJD or prion infected tissue? If so several methods of disinfection have been suggested (using the word disinfect loosely). No one is sure exactly what will inactivate this prion and the procedure has been disgusted on Histonet for routine pathology giving the latest suggested methods. I can attempt to forward some of that information to you along with the website address. Let me know if you would like the information. Most of us are alittle frightened of this one as it can take years to manifest. Pamela A. Marcum Histology/Microscopy Product Development Manager 400 Valley Road Warrington, PA 18976 Phone: 800-523-2575 Ext 167 215-343-6484 Ext 167 Fax: 215-343-0214 E-mail: pmarcum-at-polysciences.com
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Monday, April 16, 2001 9:17 AM To: Microscopy-at-sparc5.microscopy.com
Please reply directly to:
} From: {hmskaug-at-tartarus.uwa.edu.au} } } Subject: TEM } } } Hi, } } I am doing some preperation work for an assignment, and have a question. } } If a fresh unfixed piece of brain tissue (weighing approx. 1 gram) is } received in a diagnostic electron microscopy laboratory, how would I } proceed with the specimen? The brain is taken in surgery within the last 5 } min. } } If I was asked to carry out a rapid viral diagnosis on the tissue, what } precautions should I take, and how would I proceed? } I read that prions are able to survive fixation. What then? } } A respond is very much appreciated. } } Thank You ! } } Sincerely, } } Hege Skaug
Greg Erdos Assistant Director Biotechnology Program Ph. 352-392-1295 University of Florida Fax 352-846-0251 PO Box 118525 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
To all colleagues who supply standards for EDS work I need a standard sample of known thickness to check the accuracy of my TEM/EDS system during standardless quant. I need to include light elements (Z {11) as I have a Moxtek SUTW window on my detector. I guess an amorphous glass sample that contains both nitrogen and oxygen with with a very thin layer of carbon on it to prevent charging would do the trick. Can anybody out there supply one? Thanks.
Alan Fox
Dear Alan, I can't get you a standard with all the qualifications you want, but I do have some standards prepared by Chuck Fiori. The matrix is a lithium borate glass, and there are several blocks with various elements evenly dispersed in the matrix. The down side is that you will have to melt the glass, prepare thin specimens--by blowing with a platinum straw--break off pieces from the bubble, and put them on (or in) a suitable grid. I have found that folding grids work well. Of course, you will have to measure the thickness after you prepare the specimen. Anyway, it is amorphous glass with light elements, including oxygen. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
We are trying out the suggestion of using Nile Red to increase fluorescence. This seems very promising at this stage.
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
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We are looking at purchasing a stereo microscope with fluorescence capabilities primarily to look at GFP fluorescence in plant specimens. Does anyone have any particular comments on the relative merits of the systems produced by the major microscope manufactureres (Leica, Nikon, Olympus and Zeiss). We also have a dichotomy amongst users on whether to provide film or digital cameras for recording - my own preference is more on the digital side but again has anyone any comments or suggestions.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
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This is a cry for help on behalf of an unwell EDS detector that has been having some problems. It has been behaving very strangely lately but does not know who to turn to....
For the past six months it has been suffering short-lived episodes of low energy tails accompanied by some visible peak broadening most apparent at the low energy end of the spectrum. I am aware of previous discussions about possible causes of tails and peak broadening but I think the unusual thing in this case is that the problem only lasts for the first few minutes after the SEM beam is turned on and the detector is first exposed to X-rays for the day. It then gradually goes away and all aspects of the detector's behavior return to normal again. The tails won't return again that day once the SEM has been used.
This is a 10mm2 127eV Noran "Pioneer" detector with an atmospheric thin window hooked up to a Voyager IV with digital pulse processing. We leave the whole system running all of the time. It is attached to a Jeol SEM and we normally use its airlock to change samples which means that the crystal/window do not see daylight/atmospheric pressure very often. We routinely use the "slowest" highest resolution pulse processor settings. Except for this short-lived problem everything else about the system is as good as it ever was when we first installed it and ran the pulse processor setup routines a few years ago. As far as we can tell there is no *detectable* buildup of ice inside the detector (no phantom oxygen peaks etc) and there is only a small amount of visible oil contamination on the window.
It takes 12 hours or more of idleness before the tails re-appear, where then it can take 10 to 20 minutes exposure to X-rays for them to go away again. It takes less time for the tails to go away if the beam current is temporarily ramped up high enough to "saturate" the system with enough counts to approach 100% dead time. The only other unusual features that we have observed after the detector has been idle for a while are that the idle dead time indication fluctuates wildly from zero to ~50% or more from reading to reading (two second sampling periods) instead of the usual 10-20 percent, and the idle Detects and Converts are only a few counts/sec instead of the normal several tens of counts/sec. After a good dose of X-rays these parameters return to their normal idle values again also.
There is another piece to this puzzle. Last week we gave the detector a "photon enema" with a flashlight while examining the window and surrounds for anything unusual and that had the same effect in curing the problem as using a high beam current to generate X-rays.
So does anybody out there have some clues about this -- a cause, a cure, or even the physics of it? I've got the impression that it must be something electronically weird in relation to the crystal, such as some sort of an excess charge accumulation when it is idle for a long time which then gradually dissipates when the crystal starts responding to X-ray photons again? Could this effect occur if the bias was just slightly wrong by a few volts? How about "leakage current" or FET problems? I haven't got the foggiest idea really because the zero position and peak energies are always ok.
Very strange?
Concerned, Australia.
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
Ian, I have no specific comments with the new breed of fluorescent stereo's. However, in going through this a decade ago with "conventional" stereos, I found that different scopes though all well designed ranked differently with different specimens. The objects that we biologists stare at are odd optically. I suggest that you get demos and inspect the favorite objects in your departement that will be viewed under the scope. I found one stereo whose darkfield far outshone the others for arabidopsis roots, though there is no "rational" reason for this, and nothing to say that this scope is "better" in general.
A key feature to insist on is a shunt with 100% of the light to the phototube. In the old days most stereos did not do this, perhaps with fluoresecence things have changed, but you want all the light going for the captured image, whether it is film or pixels.
For image capture, going directly to a color slide can be handy. On my stereo, we trade off between video and film quite a lot. Our 35 mm camera back is no big deal (that is, just a cameraback on a tube) and adding that capability did not add much to the cost. You can of course buy much fancier and expensive 35 mm controllers but my point is you don't have to.
Hope this helps, Tobias
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Question: How long should it take for agar to conjeal after I pour it into a petri dish?
Dear Billone, Way back when I was pouring agar for immunodiffusion plates, I found that it congealed almost immediately. I had some trouble keeping it liquid to get a good smooth surface. Since that applies to the specific concentration of agar I used (I forget, but I think it was ~1%) and the temperature it was heated to (~40 C, I think), and since that was before global warming, YMMV. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Dear Ian, The recommendation from Tobias to compare before buying is always sound. We've been using a Leica MZ-12 epi-fluorescent stereomicroscope for about 2 years. GFP is only part of its use, we do a lot of microinjection and of late have been using it with vitally stained zebrafish larvae. There are a number of practical considerations. All of the mfrs. originally had limited ability to change wavelengths, the original Leica required removing the eyepieces and camera adapter as a unit, then removing screws to withdraw the filterset. Their current FL-III system has a rotating ring holding several filtersets. Filter combinations may be removed and inserted easily with it. We shuffle about 7 different sets between the filter ring, which holds about 6 (and we keep one open for brightfield). Don't ever think you will never need more than just GFP.
Adding a camera tube and fluorescent lamp housing makes the scope head very heavy. We have ours on a double arm boom (Diagnostics Inst.) to maneuver micromanipulators beneath it, and it still vibrates when you touch it. The focus must be kept tight. I just bought a lab jack stand to support our samples and to provide fine focus. Pay attention to flex in the manner in which the scope head is attached to the focus mechanism, and to the boom.
With fluorescence, consider the NA of the lenses, as that will determine whether the fluorescence is visible. We use a .8X/.05 NA for general use and long working distance with microinjection equipment. But the 1X/.125 allows us to see GFP transfected neuronal processes. A new 1.6X/.2 lens provides several times greater intensity allowing some projects to be done quickly with the stereoscope instead of having to be transferred to a compound scope.
I'd be interested in your digital camera decision. The weight issue limits what we can install. Weve been sticking with a no-name single chip color camera, which is very lightweight. The resolution is fair, and it is surprising light sensitive. The users who prefer it are viewing the color output on a passthrough monitor, then capture monochrome. The color is essential when doing multi-label injections. Color capture resolution from it is dreadful, but the monochrome is decent resolution. Other color cameras have been too heavy, inducing big vibrations when you touch the focus, or have not been sensitive enough. A Nikon Coolpix 990 is terrific for brightfield, but lacks sensitivity for fluorescence. You can get pretty good at guesstimating exposure times for 1-8 second exposures of dim fluorescence. But, the chip noise isn't worth it. And, if you need real-time imaging of low-light images for any reason (there are several reasons you might) then the Coolpix won't work. The final straw with the Coolpix, and some other cameras, is that we use some far-red dyes, like DiD, to which the Nikon is blind at working concentrations.
Regards, Glen
} } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } We are looking at purchasing a stereo microscope with } } fluorescence capabilities primarily to look at GFP fluorescence in } } plant specimens. Does anyone have any particular comments on } } the relative merits of the systems produced by the major } } microscope manufactureres (Leica, Nikon, Olympus and Zeiss). } } We also have a dichotomy amongst users on whether to provide } } film or digital cameras for recording - my own preference is more on } } the digital side but again has anyone any comments or } } suggestions. } } } } Thanks } } } } Ian } } } } } } Ian Hallett } } HortResearch } } Mt Albert Research Centre } } Private Bag 92 169 } } Auckland, New Zealand } } Fax 64-9-815 4201 } } Telephone 64-9-815 4200 } } EMail ihallett-at-hortresearch.co.nz } } }
-- Glen MacDonald UW Core for Communications Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156
Alan - The National Institute of Standands and Technology (NIST) sells a well-characterized Mg-Si-Ca-Fe-O glass thin film which is intended as a standard for EDS calibration in the TEM. The glass is supported by a 20 nm carbon thin film and its thickness has been measured by profilometry. I routinely use this standard along with other mineral standards for EDS calibration in the TEM and over the years have found it to be a very good standard indeed.
NIST can be contacted at (301)975-6776.
Dave
Dave Joswiak Dept. of Astronomy, 351580 University of Washington Seattle, WA 98195 (206)543-7702
On Mon, 16 Apr 2001, Alan Fox wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all colleagues who supply standards for EDS work } I need a standard sample of known thickness to check the } accuracy of my TEM/EDS system during standardless quant. I need to } include light elements (Z {11) as I have a Moxtek SUTW window on my } detector. I guess an amorphous glass sample that contains both nitrogen } and oxygen with with a very thin layer of carbon on it to prevent } charging would do the trick. Can anybody out there supply one? Thanks. } } Alan Fox } } } Professor Alan G. Fox BSc PhD CEng FIM } Director, Center for Materials Science and Engineering } Naval Postgraduate School } Monterey } California 93943 } USA } } Tel (831) 656 2142 (work) } (831) 657 9239 (home) } Fax (831) 656 2238 } } } }
Hi, Your agar should be setting up almost immediately. I usually use a concentration of 1-2% for my purposes. One imperative point is that the agar needs to be completely dissolved. This may sound like a trivial comment, but I know that when I first started working with agar I made the mistake of not heating the solution hot enough for a long enough time and wondered what was wrong. If you have access to an autoclave, autoclave it for about 15 minutes. If not, heat it on a hot plate (preferably with a stir bar, but you can swirl it periodically if you don't have a stir bar) until the agar is completely dissolved and you have a nice, clear solution. Be careful, because agar can suddenly start to boil and be up and over the top of your flask before you realize it. Let me know if you need any more help. Good luck, Kristen
At 04:38 PM 4/16/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu
In a message dated 04/17/2001 8:23:01 AM US Mountain Standard Time, glenmac-at-u.washington.edu writes:
{ { Adding a camera tube and fluorescent lamp housing makes the scope head very heavy. We have ours on a double arm boom (Diagnostics Inst.) to maneuver micromanipulators beneath it, and it still vibrates when you touch it. The focus must be kept tight. I just bought a lab jack stand to support our samples and to provide fine focus. Pay attention to flex in the manner in which the scope head is attached to the focus mechanism, and to the boom. } }
Glen's message is full of great advice. Regarding the point mentioned above, this is indeed crucial as you start stacking photo tubes, ergo heads, cameras, etc. onto the stereomicroscope. You can easily overwhelm the load limits of the focus drive and the gearing, and the scope will start to "droop" so that you have to continually bring the scope back into focus.
I don't know about other scopes, but Leica has a tension upgrade kit that can be installed in the focus drive unit for these situations. It makes the focus much tighter. This upgrade is for the MZ FL III which is configured (usually) on a transmitted light base with a focus post and focus drive attached to the base. As far as I know, there is no such kit if you decide to put the scope on a boom stand or a swinging arm stand. In that case, you would have to deal with the tightness of the articulated joints on the stand.
Anyway, if you order the MZ FL III stereofluorescence scope in the usual configuration, Leica has a cure for the "droopy scope" syndrome if it gets too heavy. For details on the upgrade you can contact Leica Customer Service at 1-800-248-0123. Or contact me and I will put you in touch with our Service Engineers who have installed several of the tension upgrade kits. One qualifier: installing the kit and the springs requires disassembling the focus drive unit. A factory trained service guy should definitely do this.
Good luck!
Best regards,
Bob Chiovetti GTI Microsystems, Inc. Leica Exclusive Regional Dealer Southwestern United States
I'm going out on a limb here, because it is not a particular area of expertise of mine, but I've not seen it in the discussion. Experts in microscopy of radiation sensitive materials are much more familiar with these issues (Linn Hobbs - can you help us here?)
I'm surprised that no-one has pointed out a very important difference between exposure of films to photons and electrons. Film exposed to photons exhibits a logarithmic response - that is, D varies as log(exposure). For electrons, this is not the case, but instead, the density is linear with exposure. This arises because individual grains require exposure to many photons to make them "developable", whereas a single electron will *COMPLETELY* expose many grains. For brevity, I will not explain how this changes the exposure characteristics, but it does.
A second consequence of this difference is that while change of development can change the threshold exposure for development of grains sensitised by light, grains exposed to electrons are either fully sensitized or are completely virgin, and there is *FAR* less scope to change the image characteristics by changing development of electron-exposed emulsions (though there is some, for Kodak SO163, for example).
This makes correct exposure much more critical for electron images, and makes them much more prone to overexposure. With light, and area with D of 4.0 has received 10,000 times more light than an area with D of 1.0. In an electron image the area with D of 4.0 has received 4 times more electrons than the area with D of 1.0. Do negatives really have D's above 4? Certainly they can when exposed to electrons.
There are other consequences. For example, the contrast (as we usually define it as the difference in density between different areas), which is independant of exposure on the linear portion of a film's response for light (as explained by Gary Gaugler), is, in the case of electrons, a linear function of the exposure. Underexposure leads to loss of contrast. This is why images of radiation-sensitive materials are taken at low magnification - it is the only way to maintain enough exposure of the emulsion to give acceptable contrast, without increasing the electron dose on the sample. Incidentally, the limit on information in such images is probably not the "grain size" of the emulsion, but the shot noise due to the finite number of electrons used to generate the image.
There is much more to this topic - and I have simplified what I have said.
Kristen is right on, especially with the safety issues. The completely dissolved agar should not look like it has sand in it when you swirl the flask. Be careful when swirling; if it is near the poiling point, it will foam up over the top before you can set it down. It can cause nasty burns like hot oil because it is thick.
Agar is a complex polysaccharide extracted from algae (several genera in the Rhodoophyceae). It melts somewhere around 96 degrees C and solidifies around 36 degrees C. I can't quite remember exact temperatures. Thus, to answer your solidification question, it will solidify when it gets to about body temperature. The length of time that takes after pouring will depend on how hot it was when it was poured, how thick your pour it, and how cool the Petri plate and room are.
Address at bottom if you have further questions.
On Tue, 17 Apr 2001, Kristen Lennon wrote:
} Date: Tue, 17 Apr 2001 11:48:59 -0500 } From: Kristen Lennon {kalen-at-iastate.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Ask-A-Microscopist:Agar } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } Your agar should be setting up almost immediately. I usually use a } concentration of 1-2% for my purposes. One imperative point is that the } agar needs to be completely dissolved. This may sound like a trivial } comment, but I know that when I first started working with agar I made the } mistake of not heating the solution hot enough for a long enough time and } wondered what was wrong. If you have access to an autoclave, autoclave it } for about 15 minutes. If not, heat it on a hot plate (preferably with a } stir bar, but you can swirl it periodically if you don't have a stir bar) } until the agar is completely dissolved and you have a nice, clear solution. } Be careful, because agar can suddenly start to boil and be up and over the } top of your flask before you realize it. } Let me know if you need any more help. } Good luck, } Kristen } } At 04:38 PM 4/16/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Email: wonger-at-allover.com } } Name: Billone } } } } Organization: Murphy } } } } Education: 6-8th Grade Middle School } } } } Location: San Jose, California } } } } Question: How long should it take for agar to conjeal after I pour it into } } a petri dish? } } } } --------------------------------------------------------------------------- } } Kristen A. Lennon, Ph.D. } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } 515-294-8854 } kalen-at-iastate.edu } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I have been trying to locate a point source bulb for my Durst enlarger, It is GE bulb BHD it is a single contact bayonet type that is a 100 W , 20V bulb , which I have been told is an old microscope bulb that has been discontinued with no replacement noted. If anyone knows of or has a source where I can get a suitable replacement or have that bulb, please let me know.
Thanks,
Michael Pidgeon Keck School of Medicine at USC Dept. of Cell & Neurobiology
I have been trying to locate a point source bulb for my Durst enlarger, It is GE bulb BHD it is a single contact bayonet type that is a 100 W , 20V bulb , which I have been told is an old microscope bulb that has been discontinued with no replacement noted. If anyone knows of or has a source where I can get a suitable replacement or have that bulb, please let me know.
Thanks,
Michael Pidgeon Keck School of Medicine at USC Dept. of Cell & Neurobiology
Hello Everyone, I've written some software in Perl that may be of use in laboratory facilities. We use it to schedule and monitor usage of our TEM. It simulates the old sign up sheet on the wall idea, but on the web. It has a modest amount of security to control which users can sign up onto the schedule. It also keeps a searchable archive of your past schedules for history and billing information.
I've tried to make the installation as automated as possible. You'll need to have an account on a server where you can run cgi scripts, or your own linux/sun/irix/windows nt server. You'll also need to have Perl installed where you want the software to run. Please send me comments if you have any difficulties with setting up or running the software. I'm at version 1.2 which hopefully shouldn't have too many bugs.
The link for the software is at: http://wilfred.berkeley.edu/~gordon/www-sched I ask for a small fee for our lab for the software which is explained on the web page. Gordon Vrdoljak.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Thanks to everyone who replied concerning my request for an EDS standardless quant. calibration standard for the TEM. NIST indeed do sell a mineral glass standard for just this purpose and I am looking into purchasing one of these. Thanks again for your help.
I have done some very nice preps using the Kleinschmit method. The procedure is a bit lengthy so please contact me off line and I would be more than happy to give it to you...It has some modifications but it works very nicely.
Sincerely, Maria
Maria Fazio-Zanakis AIM - TEM Laboratory Unilever Research, U.S. 45 River Road Edgewater, NJ 07020 1-201-840-2287 Maria.Fazio-Zanakis-at-unilever.com
Try Bulbtronics. Sorry, I don't know address, but I think they have a web site. They have all sorts of odd bulbs. Good luck.
On Tue, 17 Apr 2001, Michael Pidgeon wrote:
} Date: Tue, 17 Apr 2001 13:40:09 -0700 } From: Michael Pidgeon {pidgeon-at-hsc.usc.edu} } To: MS listserver {Microscopy-at-sparc5.microscopy.com} } Subject: Help locating replacment bulb } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello All. } } I have been trying to locate a point source bulb for my Durst enlarger, } It is GE bulb BHD it is a single contact bayonet type that is a 100 W , } 20V bulb , which I have been told is an old microscope bulb that has } been discontinued with no replacement noted. If anyone knows of or has } a source where I can get a suitable replacement or have that bulb, } please let me know. } } Thanks, } } Michael Pidgeon } Keck School of Medicine at USC } Dept. of Cell & Neurobiology } } 323-442-1862 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I will send you separately details of the method that I used on bacterial plasmids about 10-11 years ago (the method includes the preliminary stages as well as the technique). I can't send this to the list because the printed schedules need to be sent as attachments.
What I discovered was that it took several goes to get it right but the most important thing to avoid was 'surface active agents' which prevent a good film from spreading. Real 'double distilled water' is essential (not de-ionised or single distilled) and it should be stored in glass containers with no lubricants or plastic components (ground glass bottles or aluminium foil to prevent contact with plastic caps). The next was cleanliness - all glassware needs to be thoroughly soaked in a cleaning agent and then carefully rinsed with no use of detergents (I think some methods recommend chromic acid, but I found that a mixture of 2N (2M) nitric and 2N (1M) sulphuric acids were sufficient and a bit safer - NB obviously take care adding acids to water and do in a fume hood). Finally the 'hyperphase' containing the DNA should be mixed only a minute or 2 before use. Obviously to avoid contaminants it is based to keep a stock of chemicals just for this technique - but there aren't that many and they aren't particularly expensive.
Useful equipment would include a teflon 'Langmuir trough' to perform the spread (although I found that a square plastic dish worked well if thoroughly washed) and a rotary shadowing stage for the vacuum coater (although I just shadowed from two different angles at 90 deg of rotation - it takes longer because you have to return to air twice).
Once the technique was perfected I managed to get several dozen undergraduates to prepare DNA spreads of their plasmids, so it seemed like a fairly robust method where the important bit was the preparation before the DNA spread technique and a bit of practice. I found that in order for the students to get a representative number of plasmids it was best to photograph at about 10k and enlarge by 6x to 10x. I also found that the DNA was easiest to spot by increasing on-screen contrast in the microscope (smallest objective aperture and as low as 40kv electrons).
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
Michael Jarnik wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I would need to prepare DNA for TEM using spreading/shadowing in } Cytochrome C films. My pilot experiments using just plasmid DNA and the } Lang & Mitani method (Biopolymers, 9, p.373, 1970) worked rather poorly } and I would like to hear from people with some experience in this } method. What are the critical points here? Purity of water/chemicals, } Cyt C concentration, time? Any hints would be appreciated. } } Thanks for help, } } -- } Michael Jarnik
Michael, The contact for Bulbtronics is 1-800-654-8542 or www.bulbtronics.com. They have a wide range of bulbs, including a 12 page catalog on just bulbs for microscopes.
I also have a file on a company called Bulb Direct. 1-800-772-5267 www.bulbdirect.com. They also have an extensive catalog.
I have no financial or other interest.............
Don Marshall
} From Microscopy-request-at-sparc5.microscopy.com Tue Apr 17 16:42:12 2001
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
} } Hello All. } } I have been trying to locate a point source bulb for my Durst enlarger, } It is GE bulb BHD it is a single contact bayonet type that is a 100 W , } 20V bulb , which I have been told is an old microscope bulb that has } been discontinued with no replacement noted. If anyone knows of or has } a source where I can get a suitable replacement or have that bulb, } please let me know. } } Thanks, } } Michael Pidgeon } Keck School of Medicine at USC } Dept. of Cell & Neurobiology } } 323-442-1862 } }
They have an awesome selection of all sorts of bulbs at very good prices.
gary g.
At 01:38 PM 4/17/2001, you wrote:
} Hello All. } } I have been trying to locate a point source bulb for my Durst enlarger, } It is GE bulb BHD it is a single contact bayonet type that is a 100 W , } 20V bulb , which I have been told is an old microscope bulb that has } been discontinued with no replacement noted. If anyone knows of or has } a source where I can get a suitable replacement or have that bulb, } please let me know. } } Thanks, } } Michael Pidgeon } Keck School of Medicine at USC } Dept. of Cell & Neurobiology } } 323-442-1862
your argument about film density being linear with exposure for electrons came somewhat as a surprise. As you said, a single electron will *COMPLETELY* expose many grains. So, if the next electrons hits close to that area, some of the grains that would normally become developable are already completely exposed, thus the density of the film can not double and thus the response of the film is not linear.
There is a very simple test for the linearity of film if you have access to a field emission TEM with a biprism, or if you have a clean two beam case for high resolution. If you do a Fourier transform of just a cos-pattern, you should see two peaks in the Fourier transform. You can look this up in any handbook on Fourier transforms. If you do the same with a cos-pattern recorded on film, you will see plenty of higher order peaks in Fourier space. If you do this with a CCD camera, you will almost have to saturate the pixels to detect higher order terms. CCD cameras are, contrary to film, extremely linear.
If you have access to a film with a linear response, I would be eager to learn about it. Otherwise, for more details on my arguments, I would point to "Density Correction of Photographic Material ....", Ultramicroscopy, 55 (1994) 75-90. It gives examples of several films tested, the equations describing the behavior and tips how to linearize the data from film with software once the data are digitized.
With best regards,
Edgar Voelkl
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
--
________________ Dr. Edgar Voelkl Senior Development Staff Member ORNL Bldg 4515, MS 6064 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
Agar melts at 95 C and sets at ~40 C. If one melts agar (1-2%) directly on a hot plate, it can boil over when the temperature gets too high. However, if it is melted in a hot water bath, it will not boil over because the temperature will never get higher than 100 C.
How long does it take to set? It depends on the temperature of liquid agar when you pour it. If you pour 50 C agar, it will set immediately.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } "William F. Tivol" {wft03-at-health.state.ny.us} 04/17 8:48 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Email: wonger-at-allover.com Name: Billone
Organization: Murphy
Education: 6-8th Grade Middle School
Location: San Jose, California
Question: How long should it take for agar to conjeal after I pour it into a petri dish?
Dear Billone, Way back when I was pouring agar for immunodiffusion plates, I found that it congealed almost immediately. I had some trouble keeping it liquid to get a good smooth surface. Since that applies to the specific concentration of agar I used (I forget, but I think it was ~1%) and the temperature it was heated to (~40 C, I think), and since that was before global warming, YMMV. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
For electrons, this is not the case, but instead, the density is linear with exposure.
The only thing I will add to your consise explanation is that the density is linear with exposure only over a limited portion of the total dynamic range. The best films are linear for ODs from 0 to ~2, whereas the dynamic range is ~4 for these films. If one wants to determine the electron dose for the darker areas of the film, one must take a series of exposures to known doses of electrons and produce a curve of OD vs dose. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
} Does anyone know of service labs on the West coast for Amray } instruments? I would appreciate knowing if any exists.
A list of independent SEM service providers can be found at "www.jcnabity.com\service.htm".
Note that in some cases, these companies will service microscopes that are a significant distance from the company location. If a service area is not listed, you should contact the company to find out their service area.
If anyone has any additions or modifications that can be made to this list, please let me know.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Sorry, Tony is correct about the linearity of film, at least after digitization. We have done this test many times, and provided the film is not grossly overexposed, the average exposure time and digitized counts are a straight line. (There is a small offset which you can correct for and is probably associated with our microdensitometer electronics rather than real.) Of course, if you push the film developing, you probably lost the linearity.
On Wed, 18 Apr 2001, Dr. Edgar Voelkl wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Tony Garratt-Reed, } } your argument about film density being linear with exposure for } electrons came somewhat as a surprise. As you said, a single } electron will *COMPLETELY* expose many grains. So, if the next } electrons hits close to that area, some of the grains that would } normally become developable are already completely exposed, thus the } density of the film can not double and thus the response of the film } is not linear. } } There is a very simple test for the linearity of film if you have } access to a field emission TEM with a biprism, or if you have a clean } two beam case for high resolution. If you do a Fourier transform of } just a cos-pattern, you should see two peaks in the Fourier } transform. You can look this up in any handbook on Fourier } transforms. If you do the same with a cos-pattern recorded on film, } you will see plenty of higher order peaks in Fourier space. If you } do this with a CCD camera, you will almost have to saturate the } pixels to detect higher order terms. CCD cameras are, contrary to } film, extremely linear. } } If you have access to a film with a linear response, I would be eager } to learn about it. Otherwise, for more details on my arguments, I } would point to "Density Correction of Photographic Material ....", } Ultramicroscopy, 55 (1994) 75-90. It gives examples of several films } tested, the equations describing the behavior and tips how to } linearize the data from film with software once the data are } digitized. } } With best regards, } } Edgar Voelkl } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Colleagues- } } } } I'm going out on a limb here, because it is not a particular area of } } expertise of mine, but I've not seen it in the discussion. Experts in } } microscopy of radiation sensitive materials are much more familiar with } } these issues (Linn Hobbs - can you help us here?) } } } } I'm surprised that no-one has pointed out a very important difference } } between exposure of films to photons and electrons. Film exposed to } } photons exhibits a logarithmic response - that is, D varies as } } log(exposure). For electrons, this is not the case, but instead, the } } density is linear with exposure. This arises because individual grains } } require exposure to many photons to make them "developable", whereas a } } single electron will *COMPLETELY* expose many grains. For brevity, I will } } not explain how this changes the exposure characteristics, but it does. } } } } A second consequence of this difference is that while change of development } } can change the threshold exposure for development of grains sensitised by } } light, grains exposed to electrons are either fully sensitized or are } } completely virgin, and there is *FAR* less scope to change the image } } characteristics by changing development of electron-exposed emulsions } } (though there is some, for Kodak SO163, for example). } } } } This makes correct exposure much more critical for electron images, and } } makes them much more prone to overexposure. With light, and area with D of } } 4.0 has received 10,000 times more light than an area with D of 1.0. In an } } electron image the area with D of 4.0 has received 4 times more electrons } } than the area with D of 1.0. Do negatives really have D's above 4? } } Certainly they can when exposed to electrons. } } } } There are other consequences. For example, the contrast (as we usually } } define it as the difference in density between different areas), which is } } independant of exposure on the linear portion of a film's response for } } light (as explained by Gary Gaugler), is, in the case of electrons, a } } linear function of the exposure. Underexposure leads to loss of contrast. } } This is why images of radiation-sensitive materials are taken at low } } magnification - it is the only way to maintain enough exposure of the } } emulsion to give acceptable contrast, without increasing the electron dose } } on the sample. Incidentally, the limit on information in such images is } } probably not the "grain size" of the emulsion, but the shot noise due to } } the finite number of electrons used to generate the image. } } } } There is much more to this topic - and I have simplified what I have said. } } } } Tony } } } } } } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } } * Anthony J. Garratt-Reed M.A., D.Phil. } } * MIT, Room 13-1027 } } * 77 Massachusetts Avenue } } * Cambridge, MA 02139-4307 } } * USA } } * Phone: (617) 253-4622 } } * Fax: (617) 258-6478 } } * } } -- } } } ________________ } Dr. Edgar Voelkl } Senior Development Staff Member } ORNL } Bldg 4515, MS 6064 } 1 Bethel Valley Road } P.O. Box 2008 } Oak Ridge, TN 37831-6064 } } Tel.: (865) 574-8181 } Fax: (865) 574-4913 } email: vog-at-ornl.gov } }
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sorry too :)
however I feel quite confident that I can stand behind my experimental results. And I can assure you that film developing was not pushed. In addition, when digitizing film, you get a second non-linear function involved. Right? You can check this by using a Kodak photographic step tablet no.3. They are available off the shelf (at least used to be) and are calibrated. Density range is 21 steps from 0.05 to 3.05. But, maybe your microdensitometer is already corrected for the transmittivity curve (Pixel value P = a 10**S, where a corresponds to the illumination intensity and S is the density of the film) ?
Maybe, we are just talking degrees of linearity here? So, whatever you have may be sufficient in some cases, but no in others?
Edgar
} } Sorry, } Tony is correct about the linearity of film, at least after } digitization. We have done this test many times, and provided the film } is not grossly overexposed, the average exposure time and digitized } counts are a straight line. (There is a small offset which you can } correct for and is probably associated with our microdensitometer } electronics rather than real.) } Of course, if you push the film developing, you probably lost } the linearity. } } } On Wed, 18 Apr 2001, Dr. Edgar Voelkl wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Tony Garratt-Reed, } } } } your argument about film density being linear with exposure for } } electrons came somewhat as a surprise. As you said, a single } } electron will *COMPLETELY* expose many grains. So, if the next } } electrons hits close to that area, some of the grains that would } } normally become developable are already completely exposed, thus the } } density of the film can not double and thus the response of the film } } is not linear. } } } } There is a very simple test for the linearity of film if you have } } access to a field emission TEM with a biprism, or if you have a clean } } two beam case for high resolution. If you do a Fourier transform of } } just a cos-pattern, you should see two peaks in the Fourier } } transform. You can look this up in any handbook on Fourier } } transforms. If you do the same with a cos-pattern recorded on film, } } you will see plenty of higher order peaks in Fourier space. If you } } do this with a CCD camera, you will almost have to saturate the } } pixels to detect higher order terms. CCD cameras are, contrary to } } film, extremely linear. } } } } If you have access to a film with a linear response, I would be eager } } to learn about it. Otherwise, for more details on my arguments, I } } would point to "Density Correction of Photographic Material ....", } } Ultramicroscopy, 55 (1994) 75-90. It gives examples of several films } } tested, the equations describing the behavior and tips how to } } linearize the data from film with software once the data are } } digitized. } } } } With best regards, } } } } Edgar Voelkl } } } } } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Colleagues- } } } } } } I'm going out on a limb here, because it is not a particular area of } } } expertise of mine, but I've not seen it in the discussion. Experts in } } } microscopy of radiation sensitive materials are much more familiar with } } } these issues (Linn Hobbs - can you help us here?) } } } } } } I'm surprised that no-one has pointed out a very important difference } } } between exposure of films to photons and electrons. Film exposed to } } } photons exhibits a logarithmic response - that is, D varies as } } } log(exposure). For electrons, this is not the case, but instead, the } } } density is linear with exposure. This arises because individual grains } } } require exposure to many photons to make them "developable", whereas a } } } single electron will *COMPLETELY* expose many grains. For brevity, I will } } } not explain how this changes the exposure characteristics, but it does. } } } } } } A second consequence of this difference is that while change of development } } } can change the threshold exposure for development of grains sensitised by } } } light, grains exposed to electrons are either fully sensitized or are } } } completely virgin, and there is *FAR* less scope to change the image } } } characteristics by changing development of electron-exposed emulsions } } } (though there is some, for Kodak SO163, for example). } } } } } } This makes correct exposure much more critical for electron images, and } } } makes them much more prone to overexposure. With light, and area with D of } } } 4.0 has received 10,000 times more light than an area with D of 1.0. In an } } } electron image the area with D of 4.0 has received 4 times more electrons } } } than the area with D of 1.0. Do negatives really have D's above 4? } } } Certainly they can when exposed to electrons. } } } } } } There are other consequences. For example, the contrast (as we usually } } } define it as the difference in density between different areas), which is } } } independant of exposure on the linear portion of a film's response for } } } light (as explained by Gary Gaugler), is, in the case of electrons, a } } } linear function of the exposure. Underexposure leads to loss of contrast. } } } This is why images of radiation-sensitive materials are taken at low } } } magnification - it is the only way to maintain enough exposure of the } } } emulsion to give acceptable contrast, without increasing the electron dose } } } on the sample. Incidentally, the limit on information in such images is } } } probably not the "grain size" of the emulsion, but the shot noise due to } } } the finite number of electrons used to generate the image. } } } } } } There is much more to this topic - and I have simplified what I have said. } } } } } } Tony } } } } } } } } } } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } } } * Anthony J. Garratt-Reed M.A., D.Phil. } } } * MIT, Room 13-1027 } } } * 77 Massachusetts Avenue } } } * Cambridge, MA 02139-4307 } } } * USA } } } * Phone: (617) 253-4622 } } } * Fax: (617) 258-6478 } } } * } } } } -- } } } } } } ________________ } } Dr. Edgar Voelkl } } Senior Development Staff Member } } ORNL } } Bldg 4515, MS 6064 } } 1 Bethel Valley Road } } P.O. Box 2008 } } Oak Ridge, TN 37831-6064 } } } } Tel.: (865) 574-8181 } } Fax: (865) 574-4913 } } email: vog-at-ornl.gov } } } } } } ------------------------------------------------------- } Laurence Marks } Department of Materials Science and Engineering & } Center for Transportation Nanotechnology } Northwestern University } Tel: (847) 491-3996 Fax: (847) 491-7820 } mailto:ldm-at-risc4.numis.nwu.edu } http://www.numis.nwu.edu http://www.ctn.northwestern.edu } ------------------------------------------------------- } The Other Nanotubes http://focus.aps.org/open/st12.html } Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } http://xraysweb.lbl.gov/esg/phasing/index.html
--
________________ Dr. Edgar Voelkl Senior Development Staff Member ORNL Bldg 4515, MS 6064 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
Very often it seems that film is described as non-linear, direct CCD collectors are linear, so bye-bye film. Since we did not then (and still do not have) a good CCD detector on our UHV-HREM, we investigated this in detail some years ago to get quantitative TED data from surfaces. While there are of course logarithmic terms involved, with a good drum scanner the electronics to handle this works well. If anyone wants we have calibration curves sitting on the wall over our scanner and I can send them. So long as you maintain the microdensitometer, performing occaisional calibrations, everything is fine. (A microdensitometer without reasonable TLC - GIGO.)
Both CCD camera's on the microscope and scanners have point-spread functions, and these can be important. For instance, we have a rotating drum scanner so the PSF is different along the rotation direction versus at right angles to it. I do not know what the PSF's for modern scanners are like - hence the question I raised a little time ago (but nobody had more information). You also have to have adequate sampling and similar "stuff" under control otherwise aliasing kills you.
A 2kx2k CCD is certainly competitive with film, depending upon the application. Scratches and damage from high intensities are problems for CCD's, but having an image immediately available so you can look at the power-spectrum is an advantage. However, I think the price of the TEM units has to come down before film and $2k (or even $20k) scanners become obsolete.
On Wed, 18 Apr 2001, Dr. Edgar Voelkl wrote:
} } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } Sorry too :) } } however I feel quite confident that I can stand behind my } experimental results. And I can assure you that film developing was } not pushed. In addition, when digitizing film, you get a second } non-linear function involved. Right? You can check this by using a } Kodak photographic step tablet no.3. They are available off the } shelf (at least used to be) and are calibrated. Density range is 21 } steps from 0.05 to 3.05. But, maybe your microdensitometer is } already corrected for the transmittivity curve (Pixel value P = a } 10**S, where a corresponds to the illumination intensity and S is the } density of the film) ? } } Maybe, we are just talking degrees of linearity here? So, whatever } you have may be sufficient in some cases, but no in others? } } Edgar } } } } } } } Sorry, } } Tony is correct about the linearity of film, at least after } } digitization. We have done this test many times, and provided the film } } is not grossly overexposed, the average exposure time and digitized } } counts are a straight line. (There is a small offset which you can } } correct for and is probably associated with our microdensitometer } } electronics rather than real.) } } Of course, if you push the film developing, you probably lost } } the linearity. } } } } } } On Wed, 18 Apr 2001, Dr. Edgar Voelkl wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Tony Garratt-Reed, } } } } } } your argument about film density being linear with exposure for } } } electrons came somewhat as a surprise. As you said, a single } } } electron will *COMPLETELY* expose many grains. So, if the next } } } electrons hits close to that area, some of the grains that would } } } normally become developable are already completely exposed, thus the } } } density of the film can not double and thus the response of the film } } } is not linear. } } } } } } There is a very simple test for the linearity of film if you have } } } access to a field emission TEM with a biprism, or if you have a clean } } } two beam case for high resolution. If you do a Fourier transform of } } } just a cos-pattern, you should see two peaks in the Fourier } } } transform. You can look this up in any handbook on Fourier } } } transforms. If you do the same with a cos-pattern recorded on film, } } } you will see plenty of higher order peaks in Fourier space. If you } } } do this with a CCD camera, you will almost have to saturate the } } } pixels to detect higher order terms. CCD cameras are, contrary to } } } film, extremely linear. } } } } } } If you have access to a film with a linear response, I would be eager } } } to learn about it. Otherwise, for more details on my arguments, I } } } would point to "Density Correction of Photographic Material ....", } } } Ultramicroscopy, 55 (1994) 75-90. It gives examples of several films } } } tested, the equations describing the behavior and tips how to } } } linearize the data from film with software once the data are } } } digitized. } } } } } } With best regards, } } } } } } Edgar Voelkl } } } } } } } } } } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Colleagues- } } } } } } } } I'm going out on a limb here, because it is not a particular area of } } } } expertise of mine, but I've not seen it in the discussion. Experts in } } } } microscopy of radiation sensitive materials are much more familiar with } } } } these issues (Linn Hobbs - can you help us here?) } } } } } } } } I'm surprised that no-one has pointed out a very important difference } } } } between exposure of films to photons and electrons. Film exposed to } } } } photons exhibits a logarithmic response - that is, D varies as } } } } log(exposure). For electrons, this is not the case, but instead, the } } } } density is linear with exposure. This arises because individual grains } } } } require exposure to many photons to make them "developable", whereas a } } } } single electron will *COMPLETELY* expose many grains. For brevity, I will } } } } not explain how this changes the exposure characteristics, but it does. } } } } } } } } A second consequence of this difference is that while change of development } } } } can change the threshold exposure for development of grains sensitised by } } } } light, grains exposed to electrons are either fully sensitized or are } } } } completely virgin, and there is *FAR* less scope to change the image } } } } characteristics by changing development of electron-exposed emulsions } } } } (though there is some, for Kodak SO163, for example). } } } } } } } } This makes correct exposure much more critical for electron images, and } } } } makes them much more prone to overexposure. With light, and area with D of } } } } 4.0 has received 10,000 times more light than an area with D of 1.0. In an } } } } electron image the area with D of 4.0 has received 4 times more electrons } } } } than the area with D of 1.0. Do negatives really have D's above 4? } } } } Certainly they can when exposed to electrons. } } } } } } } } There are other consequences. For example, the contrast (as we usually } } } } define it as the difference in density between different areas), which is } } } } independant of exposure on the linear portion of a film's response for } } } } light (as explained by Gary Gaugler), is, in the case of electrons, a } } } } linear function of the exposure. Underexposure leads to loss of contrast. } } } } This is why images of radiation-sensitive materials are taken at low } } } } magnification - it is the only way to maintain enough exposure of the } } } } emulsion to give acceptable contrast, without increasing the electron dose } } } } on the sample. Incidentally, the limit on information in such images is } } } } probably not the "grain size" of the emulsion, but the shot noise due to } } } } the finite number of electrons used to generate the image. } } } } } } } } There is much more to this topic - and I have simplified what I have said. } } } } } } } } Tony } } } } } } } } } } } } } } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } } } } * Anthony J. Garratt-Reed M.A., D.Phil. } } } } * MIT, Room 13-1027 } } } } * 77 Massachusetts Avenue } } } } * Cambridge, MA 02139-4307 } } } } * USA } } } } * Phone: (617) 253-4622 } } } } * Fax: (617) 258-6478 } } } } * } } } } } } -- } } } } } } } } } ________________ } } } Dr. Edgar Voelkl } } } Senior Development Staff Member } } } ORNL } } } Bldg 4515, MS 6064 } } } 1 Bethel Valley Road } } } P.O. Box 2008 } } } Oak Ridge, TN 37831-6064 } } } } } } Tel.: (865) 574-8181 } } } Fax: (865) 574-4913 } } } email: vog-at-ornl.gov } } } } } } } } } } ------------------------------------------------------- } } Laurence Marks } } Department of Materials Science and Engineering & } } Center for Transportation Nanotechnology } } Northwestern University } } Tel: (847) 491-3996 Fax: (847) 491-7820 } } mailto:ldm-at-risc4.numis.nwu.edu } } http://www.numis.nwu.edu http://www.ctn.northwestern.edu } } ------------------------------------------------------- } } The Other Nanotubes http://focus.aps.org/open/st12.html } } Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html } } } } Workshop May 17-19 2001 "New approaches to the Phase Problem" } } http://xraysweb.lbl.gov/esg/phasing/index.html } } -- } } } ________________ } Dr. Edgar Voelkl } Senior Development Staff Member } ORNL } Bldg 4515, MS 6064 } 1 Bethel Valley Road } P.O. Box 2008 } Oak Ridge, TN 37831-6064 } } Tel.: (865) 574-8181 } Fax: (865) 574-4913 } email: vog-at-ornl.gov } }
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Ed Voelkl is a good microscopist whom I like and respect but he is wrong. Of course, all detectors saturate if the signal is large enough. But the point is how does it behave in the useful detection range. When film is exposed to electrons the density increases linearly with dose - until saturation effects begin. By contrast when exposed to light, film is not linear anywhere.
If you want to get this right, please read:
The response of photographic emulsions to electrons by R C Valentine in Advances in Optical and Electron Microscopy, volume 1 Edited by R Barer and V E Cosslett Academic Press
This is one of those definitive and good articles - which is also clear and easy to read.
Alwyn Eades -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Hey, I am enjoying this discussion. Any chance to get a lunchtime debate going at the M&M meeting like we had a few years ago?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] } Sent: Wednesday, April 18, 2001 2:55 PM } To: EMNET } Subject: Exposure of film to electrons. } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } Ed Voelkl is a good microscopist whom I like and respect but he is } wrong. Of course, all detectors saturate if the signal is large } enough. But the point is how does it behave in the useful detection } range. When film is exposed to electrons the density } increases linearly } with dose - until saturation effects begin. By contrast when exposed } to light, film is not linear anywhere. } } If you want to get this right, please read: } } The response of photographic emulsions to electrons } by R C Valentine } in Advances in Optical and Electron Microscopy, volume 1 } Edited by R Barer and V E Cosslett } Academic Press } } This is one of those definitive and good articles - which is } also clear } and easy to read. } } Alwyn Eades } -- } .......... } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvania 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu }
Mary McCann's email address is: mccanns-at-tiac.net. I'm sorry for any inconvenience to those people who have tried, unsuccessfully, to reach her!
If anyone is interested in attending the meeting--you can email me directly and I can send you a newsletter or fax you the information re: program and registration.
Please include a complete mailing address and fax number.
Thanks! Peggy Sherwood Corresponding Secretary, NESM -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
H. Frieser and E. Klein, Z. Angew. Phys. 10 (1958) 337.
H. Frieser, E. Klein and E. Zeitler, Z. Angew. Phys. 11 (1959) 190.
E. Zeitler, Ultramicroscopy 46 (1992) 405.
E. Voelkl, F. Lenz, Q. Fu and H. Lichte, "Density correction of photographic material for further image processing ....", UM 55 (1994) 75-89.
against:
R C Valentine, in Advances in Optical and Electron Microscopy, volume 1 Edited by R Barer and V E Cosslett, Academic Press
that's 4:1 :)
Edgar
P.S.: So far, the argument remains: a single electron "A" will completely expose many grains. So, if the next electron "B" hits close to the same area, some of the grains are already completely exposed through electron "A", thus less grains become fully developed with electron "B". Therefore, the density of the film can not double and thus the response of the film is non-linear.
At 2:55 PM -0400 4/18/01, Alwyn Eades wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
--
________________ Dr. Edgar Voelkl Senior Development Staff Member ORNL Bldg 4515, MS 6064 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
Since this seems to be a lively thread, I would like to make the following suggestion:
Most of us have an electron microscope, and I would like to suggest the following experiment to test if your film is linear or not. It is a simple, two-step test.
1) Take a high resolution image of some periodic structure such that only two beams contribute to the image, preferably with identical intensity. This will provide a cos- type imprint on your film (actually a P+cos(...) type imprint with P} 1).
2) Develop the film and look through it at a small light source, e.g., a halogen light. If necessary, tilt the film a little to decrease fringe spacing for better separation of the diffraction orders.
If you have a true linear film, no higher diffraction orders will be visible. The two diffracted beams will be quite colorful like a rainbow. If you see more than two diffracted beams, your film has a non-linear response (all of those I have ever seen do (disclaimer: the last time I have dealt with film was 1992)).
BTW,
I like Scotts suggestion of a lunchtime debate going at the M&M meeting. Monday is out with me, but any other day is fine.
Edgar
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
--
________________ Dr. Edgar Voelkl Senior Development Staff Member ORNL Bldg 4515, MS 6064 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
Amray has a list of third party repair outfits. I got it from Amray. I can fax it to you or call Amray and get it direct.
gary g.
At 11:20 AM 4/17/2001, you wrote:
} Does anyone know of service labs on the West coast for Amray } instruments? I would appreciate knowing if any exists. } Thank you. } } Franklin Bailey } University of Idaho } Moscow, ID 83844-2204 } jfb-at-uidaho.edu
Quick and dirty way to check the linearity of the whole acquisition process (film + scanner) is to take multiple exposures on a film by gradually exposing its area. I've done this by using the selected area aperture. First position the aperture so that only a small stripe of the film to be exposed. Make an exposure. Then move the aperture to open a larger area (next stripe). Expose ... Use a relatively low dose so that after 20 or 30 stripes the film to be exposed to twice the dose you use in your experiments. After scanning the film make a "line projection". Plotting the density vs. stripe number (dose) will show you the linearity region of your acquisition system (film + scanner). A quick way to check the Modulation Transfer Function (MTF) of the acquisition system is to take an exposure on the film without specimen (beam only). Theoretically the Fourier transform of the scan should show a constant amplitude over all spatial frequencies (Shot noise = white noise). Average rotationally the amplitude of the Fourier transform - this is the MTF.
Does anyone know of a digital imaging system for scanning EM? I know that CCD imaging systems for TEMs have been discussed in the group before, but I don't recall whether SEMs have been discussed. Perhaps someone could tell me if this subject has been archived, or perhaps someone has a summary. Vendors should also feel free to respond directly to me off-list.
Someone has asked me for information on a digital image acquisition system for SEM (they currently have a screen with a Polaroid camera attachment). If anyone has recommendations for such a digital acquisition system, please contact me with the details.
Thanks very much in advance.
Bob Chiovetti GTI Microsystems, Inc. Leica Exclusive Regional Dealer Southwestern United States rchiovetti-at-aol.com
Just let you know that there will be two short courses on food microscopy in Minneapolis on May 13 as part of the Food Structure & Functionality Symposium.
Course 1 is aimed at researchers interested in specific localisation techniques as a tool for understanding structure-function relationships in foods. Course 2 gives a grounding in light microscopy technques (optical contrast, staining) used for the identification of food materials and contaminants (glass, fibres, plastics etc) and is aimed at quality control/research & development personnel.
Details can be found at: http://www.aocs.org/meetings/am2001/foodstr.htm
Both courses offer practical tuition by internationally recognised speakers.
Let me know if you are interested, or you can book online at: https://www.aocs.org/meetings/am2001/regshort.htm
Regards Mark
Mark Auty Dairy Products Research Centre Moorepark Fermoy Co. Cork Ireland
tel +353 25 42447 fax +353 25 42340 mauty-at-moorepark.teagasc.ie
Just let you know that there will be two short courses on food microscopy in Minneapolis on May 13 as part of the Food Structure & Functionality Symposium.
Course 1 is aimed at researchers interested in specific localisation techniques as a tool for understanding structure-function relationships in foods. Course 2 gives a grounding in light microscopy technques (optical contrast, staining) used for the identification of food materials and contaminants (glass, fibres, plastics etc) and is aimed at quality control/research & development personnel.
Details can be found at: http://www.aocs.org/meetings/am2001/foodstr.htm
Both courses offer practical tuition by internationally recognised speakers.
Let me know if you are interested, or you can book online at: https://www.aocs.org/meetings/am2001/regshort.htm
Regards Mark
Mark Auty Dairy Products Research Centre Moorepark Fermoy Co. Cork Ireland
tel +353 25 42447 fax +353 25 42340 mauty-at-moorepark.teagasc.ie
Hi, I would like to get in contact with anyone currently using a JEOL-220A SEM. I particularly want to know if it is possible to add a digital image acquisition system to this SEM. Our lab has a chance to acquire a 220A which has less than 200 hours of use and was under service contract before going into storage 4 years ago. Any and all information would be appreciated. Thanks.
Tom Bargar EM Lab U. Neb. Med. Ctr. phone (402)-559-7347 tbargar-at-unmc.edu
The Biomaterials Science Group Department of Oral and Dental Science University of Bristol in collaboration with Glaxo SmithKline
Postdoctoral Research Assistant
in the Biomaterials Science Group on the project Interaction Mechanisms of Polymers at Interfaces of Mineralised Tissues
The research area involves the study of the physical and chemical properties and interaction mechanisms of different polymers at interfaces of mineralised tissues. You will have recently been awarded a PhD in an appropriate field and will ideally have experience in scanning probe microscopy (AFM) of biological materials and other analytical techniques and an interest in medical research. You will work in Dr. Jandts group and interact with scientists at Glaxo Smith Kline. The University of Bristol is one of the leading research universities in the UK and provides an outstanding scientific training environment to enhance your qualification. The group is involved in exciting, interdisciplinary projects and maintains appropriate state of the art instrumentation. There exist opportunities for additional interactions with clinical scientists and other centres at the university. We are looking for a dynamic and exceptionally well-qualified postdoctoral researcher who can interact effectively in an international and interdisciplinary team. The appointment will be on a Research Assistant 1A scale with a salary range of # 16775 to # 20465. This is a full time appointment and initially for one year. Applicants should include a short CV, stating research experience and interests, publication list and addresses of two referees. The review of applications will start 24 May 2001 and will continue until the post has been filled. Informal inquiries can be directed by email to Dr. K. D. Jandt (K.Jandt-at-bris.ac.uk), Senior Lecturer in Biomaterials, University of Bristol
Formal applications quoting the reference number 7401 should be directed to
The University of Bristol Recruitment Office Bristol, BS8 1TH United Kingdom
----------------------------------------------------------------- Dr. rer. nat. Klaus D. Jandt Senior Lecturer in Dental Materials Science and Biomaterials University of Bristol, Department of Oral and Dental Science Lower Maudlin Street, Bristol, BS1 2LY, UK Phone: +44-117-9284418, Fax: ++44-117-9284780 Internet: K.Jandt-at-bris.ac.uk WWW: http://www.dent.bris.ac.uk/Biomaterials/kdj.htm "We make Biomaterials Science work!"
I have managed to get a Varian 842 ionization gauge controller and am working on the tube itself, leaving only one component - the cable(s) - remaining. Varian wants $190 for this cable, which seems a little steep to me.
} From the connections on the controller box it looks like there is a four wire cable with an octal plug going that would go to some connector on the bottom of the tube and a separate coax connection for the collector. Are the connectors that would attach to the bottom and top of the tube readily available components and, if so, where could I buy them? Is there some place where I could simply buy the entire cable system at a better price than what Varian is asking? What is so special about five wires and a couple of connectors that warrants that kind of price?
I would appreciate information from anyone regarding the connection of ion gauge tubes to their controllers, Varian or not.
ASSISTANT DIRECTOR (TECHNICAL) Acadia Centre for Microstructural Analysis (ACMA) Competition #01-17
The Acadia Centre for Microstructural Analysis (ACMA) has recently been established to support research initiatives within the Faculty of Pure & Applied Science and provide technical services to the regional R&D community. We are seeking an individual to manage and maintain the microanalytical facility consisting of scanning and transmission electron microscopes, scanning probe microscope, FTIR spectrometer, epi-fluorescence and confocal microscopes along with associated specimen preparation equipment.
Responsibilities: management of day to day operations including training and general supervision of users, technical support and general maintenance of research equipment, technical service and consulting activities, marketing of technical services.
Qualifications: A minimum of a Masters Degree in natural sciences/engineering with courses in computer programming and electronics or an equivalent combination of education and related work experience; experience in instrumentation; some expertise with one or more of TEM, SEM, FTIR, SPM or confocal microscopy; strong communication, writing and interpersonal skills. An ability to provide general electronics and instrumentation support to the Faculty would be considered an asset.
The Assistant Director will be a full-time employee of Acadia University and the initial appointment shall be for one year, with the possibility of renewal for up to five years.
Salary Range: $39,615 to $51,645 per annum, depending on qualifications
Closing Date for Applications: June 1, 2001
Please send your resume including three letters of references to: Marian Reid, Personnel Officer Acadia University, Wolfville NS B0P 1X0 E-mail: marian.reid-at-acadiau.ca Fax: 902-585-1075
We thank all applicants in advance but advise that only those selected for an interview will be contacted. Acadia University reserves the right not to fill this position.
In accordance with Canadian immigration requirements, this advertisement is directed to Canadian citizens and permanent residents.
An equal opportunity employer, Acadia welcomes applications from qualified women and men, including African Nova Scotians, First Nations peoples, persons with disabilities, and racially visible people. Such individuals are encouraged to self-identify.
Dr. Craig Bennett Associate Professor and Acting Head Department of Physics, Acadia University and Director, Acadia Centre for Microstructural Analysis Wolfville, Nova Scotia, Canada B0P 1X0 tel. 902-585-1150 fax. 902-585-1816
Hi all! I have a quick question. I am working with purified nuclear matrix samples and need tips for the fixation / embedding process. First of all, the samples are so small, they are nearly invisible, and secondly, they are incredibly fragile, so great care must be taken during this whole process. To complicate matters, we are doing pre-embedding immunolabeling. The problem comes in after incubation in secondary (gold) antibody. We have to spin down the sample after each rinse or incubation. My concern is about the gold particles........At what RPM will they sediment? Has anyone ever encountered this dileama before? Likewise, does anyone have any tips for fixation and embedding of this kind of sample? Question # 2: Is it ok to go straight from Prop Ox to 100% resin (spurr's)? The matrix is mostly protein, so it shouldn't take long at all to infiltrate...but I am unsure about how to go about infiltrating. In the viscous resin, we won't be able to spin down the matrix, that's where the problem arises.
Any help would be greatly appreciated!
Thanks in advance, Karli Karli Fitzelle MCI Center OARDC / OSU Wooster, Ohio 44691 330-263-3828
I have a user in our EM Center who would like to have a procedure for determining the thickness of their ultrathin sections, 60-100 nm. They are using the normal color criterion of "silver" sections in the boat to select their sections, but would like to be more precise. There is a potential for doing morphometry and comparisons of particle counts between non-serial sections and sections from different specimens.
They have heard of a technique that uses small particles applied to both surfaces of the section and using tilt and geometry of the TEM stage to determine section thickness. Any details of this technique would be appreciated.
All suggestions are welcome.
John Colorado State University john.chandler-at-colostate.edu
Sample holders for JEOL 100CX-2000EXII model TEMs for sale:
Gatan hot stage sample holders, air/water cooled. Both furnaces in working order; holders include one hexnut lockring AND thermocouple controller (sorry, no power wires--lost!): --#1 in good condition, $12,000 (Gatan Power supply model #580-0300) --#2 lightly stripped hex nut assembly in furnace (doesn't tighten all the way but sample will remain in place fairly well), mA display on power supply (Gatan power supply model #628-0500) a bit jumpy but thermocouple reads temperature fine, $7000
Extra washers (Gatan part #628-0223), and hex nut tool (Gatan part #608-0005) NOT included with the above holders.
Please contact Terry K. Baker for further inquiries and negotiations: Phone 508 893 9560 baker-at-catalytic-materials.com Catalytic Materials 1750 Washington St. West Holliston Professional Park, Suite C2 Holliston, MA 01746
I would try to immobilize the stuff on some substrate that could then be carried like a piece of tissue. I am not sure what that would be in this case. Possibly a Nuclepore filter???
At 03:27 PM 4/19/2001 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greg Erdos Assistant Director Biotechnology Program Ph. 352-392-1295 University of Florida Fax 352-846-0251 PO Box 118525 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
I have been asked by one of our collegues hereas to wether there is a selective stain for viewing xylem tissue that has been embedded in resin. These are 1-7um sections viewed with a light microscope
Somewhere in the deep dark recesses (which are not accessible on a Friday afternoon or Monday morning ) I seem to remember there is one; or indeed, a publication that lists many of these such permeations.
Any help appreciated Cheers
Raymond Bennett Keith Williamson EM Unit Hort+Research Palmerston North NEW ZEALAND
_________________________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. _________________________________________________________________
} I have managed to get a Varian 842 ionization gauge controller and am } working on the tube itself, leaving only one component - the cable(s) - } remaining. Varian wants $190 for this cable, which seems a little steep to } me. } } } From the connections on the controller box it looks like there is a four } wire cable with an octal plug going that would go to some connector on the } bottom of the tube and a separate coax connection for the collector. Are the } connectors that would attach to the bottom and top of the tube readily } available components and, if so, where could I buy them? Is there some place } where I could simply buy the entire cable system at a better price than what } Varian is asking? What is so special about five wires and a couple of } connectors that warrants that kind of price? } } I would appreciate information from anyone regarding the connection of ion } gauge tubes to their controllers, Varian or not. } } Bruce Girrell
I got a similar cable from Duniway Stockroom. They were much cheaper than Varian, and the cable has worked great.
Tom
-- Thomas Mullarkey Murray email:tm8a-at-virginia.edu Thornton Hall - MSE phone:(804)982-5659 University of Virginia Fax: (804)982-5660 Charlottesville, VA 22903
Earl ----- Original Message ----- } From: "Bruce Girrell" {bigirrell-at-microlinetc.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 19, 2001 9:26 AM
Question: I am learning to do SEM now and have been doing immunolocalization (histogold) for light microscopy. I am now interested in applying this histogold immunolocalization in EM work. It appears that most immunolocalization I've seen is in TEM. I am working in SEM. Is there any advantage of one over the other besides the obvious difference that TEM looks inside the tissue and SEM looks at the surface? How come there aren't as many SEM localizations out there (that I've seen)? Thank you for your time, Karen
} Subject: EDXA, Need help with detector geometry } } } } Sirs or Madams, } } } } I am running a JEOL CX II with a Kevex mod. 3200-0018 detector/ Kevex Delta } } Class Anlyzer. } } } } I am having difficulty locating the geometric variables unique to this } } mating of scope and detector. Kevex was unable to supply the data. These } } geometric variables are used by the analyzer software (Quantex) in modeling } } and subtracting backgrounds. } } } } The variables I am unable to supply are Working Distance, Fixed Distance, } } and Azimuth. I have seen reference to a Quantex Parameters List. This } } document was shipped with the original equipment, but alas, this is a } second } } hand scope and the detector was taken from the company warehouse. } } } } Does anybody use this combination of TEM and detector or know of someone } } with this combination? Does anyone wish to share a document listing } Quantex } } parameters for different scopes with Kevex detectors? } } } } My sanity is in your hands. } } I remain humbly yours, } } } } } } Stephen Bennett } } EMSL Analytical, Inc. } } Miami, FL } } } } miamilab-at-emsl.com } }
I am sure you will find differential staining methods for xylem in there ..
Using Toluidin Blue O gives good results just by the more intense (dark blue) xylem cell walls compared to the thinner (light blue) parenchyma cell walls - but this is not a differential staining method!
Hope this helps you,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
I have a Hitachi S-800 FESEM gun bellows that has developed a small leak. Although it has been replaced, I have heard of bellows being repaired by plating with cadmium or some other metal.
Karli Greg is right. A nuclepore filter could be used to separate small specimens from reagents, which could be exchanged using a syringe. But this is only one of various options. Another option would be to encapsulate the specimens in low-melting point agarose. Then they can be handled in the same way as as tissue blocks. However, centrifugation using a low-speed centrifuge such as an eppendorff centrifuge, will easily separate the unbound gold probe from your specimens. If you are worried about this, test a sample of you colloidal gold probe in your centrifuge. If the supernatant stays pink and there is no red pellet the gold is still a sol. I would recommend that you process the tissue as you are doing, using centrifugation, until the immunolabelling procedure is completed, and at that stage embed the specimen pellet into agarose prior to dehydration and resin infiltration.
Moving your specimen direct from propylene oxide to pure resin is a recipe for specimen collapse unless your specimens are a) exceedingly small, b) very permeable to the resin. What happens is the very mobile PPO comes out of the specimen faster than the viscous resin can move into it, resulting in a volume reduction and shrinkage. You can normally get away with two or three intermediate steps and these are much easier to accomplish if your specimens are in large pieces (we're talking relative sizes here - large means visible, cubes maybe 0.2 to 1 mm in a side) Good luck Chris
----- Original Message ----- } From: "Greg Erdos" {gwe-at-biotech.ufl.edu} To: "Karli Fitzelle" {fitzelle.1-at-osu.edu} ; {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 19, 2001 9:58 PM
Hi John,
This is a simple parallax problem.
Imagine the specimen in cross section. If there are two particles one vertically above the other they are seperated by the film thickness T. Tilt the film through an angle A and in plan view the particles will seperate by a distance D. (This can also be extended to account for two particles not vertically above each other but I'll stick to the easy case for the explaination.)
Take two negatives one at zero tilt and one at tilt of A and measure the seperation D, the thickness can be calculated by T=D/sinA.
You will need to know the direction of the tilt axis for your measurements and you can see that the larger the tilt angle and the more acurately you can measure the seperation the more accurate your measurement will be. Tilt + and - A to get a more accurate result.
Now you have to think about what you are going to use to mark the surfaces as you need to be able to distinguish the markers at two different tilt angles. Crystals may go in and out of contrast making them difficult to follow. If you use spheres don't forget to subtract the diameter of the sphere (or one the sum of the two radii) from your total thickness to account for the fact that you are probably using the circumferance as the marker.
Of course the alternative is to re-embed and cross section to measure the thickness directly.
With any luck there may be some responses from people currently measuring thicknesses using this, or other techniques, with some relevant hints or comments on expected accuracy of the different methods.
Good luck, Ron
On Thu, 19 Apr 2001 13:35:43 -0600 John Chandler {chandler-at-lamar.ColoState.EDU} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a user in our EM Center who would like to have a procedure for } determining the thickness of their ultrathin sections, 60-100 nm. They are } using the normal color criterion of "silver" sections in the boat to select } their sections, but would like to be more precise. There is a potential for } doing morphometry and comparisons of particle counts between non-serial } sections and sections from different specimens. } } They have heard of a technique that uses small particles applied to both } surfaces of the section and using tilt and geometry of the TEM stage to } determine section thickness. Any details of this technique would be } appreciated. } } All suggestions are welcome. } } John } Colorado State University } john.chandler-at-colostate.edu } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
We get the bellows replaced by a local (UK) firm, typically around 250 pounds for edge welded bellows in stainless steel. This includes removing the old bellows, supply and welding in new bellows and leak testing. It usually takes a few weeks unless we are prepared to pay to interupt the work scedule. These prices are for 20-40mm dia 50-80mm long bellows.
There must be firms in the US (and most other places) to do this.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I have a Hitachi S-800 FESEM gun bellows that has developed a small leak. } Although it has been replaced, I have heard of bellows being repaired by } plating with cadmium or some other metal. } } Does anyone have any experience with this? } } Thank You, } } Earl Weltmer } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
----- Original Message ----- } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: {karenco-at-discoverymail.com} Sent: Friday, April 20, 2001 8:52 AM
Howdy Y'all,
I am looking for an inexpensive vise-style specimen holder for our SEM. It does not need to be fancy, just a flat-jawed simple device with a screw to tighten the faces of the jaws. After becoming frustrated by all the information available on the web,I'm sure someone out there could be of assistance. Please help by responding online.
Mr. Weltmer, Try finding an aircraft grade, two-part epoxy (Epoxo 88). If you have stress cracks on a vacuum bellows, mix a batch of epoxy and thinly spread over the joints and areas where you suspect a crack. Make sure to prep the area with acetone or methanol and wait for the solvent to flash off before applying the epoxy. I have repaired a vacuum manifold on a Poloron plasma asher and have not had a leak for a year now. The epoxy doesn't seem to outgass enough to bother. You may want to place the joint in a muffle furnace at low temp. after a few hours to make sure the product is fully cured.
Hi Karen, Here is a paper that deals with immunoelectron microscopy for the SEM. If you have any questions you can contact me at the e-mail or phone below.
Coller, Barry S., Kutok, J. L., Scudder, L. E., Galanakis, D. K., West, S. M . , Rudomen, G. S., Springer, K. T., "Studies of Activated GPIIb/IIIa Receptors on the Luminal Surface of Adherent Platelets: Paradoxical Loss of Luminal Receptors When Platelets Adhere to High Density Fibrinogen." J. Clin. Invest. Vol. 92, pp. 2796-2806, 1993
karenco-at-discoverymail.com wrote: }
} } Email: karenco-at-discoverymail.com } Name: karen chamusco } } Organization: university of florida } } Education: Graduate College } } Location: gainesville, fl } } Question: I am learning to do SEM now and have been doing } immunolocalization (histogold) for light microscopy. I am now } interested in applying this histogold immunolocalization in EM work. } It appears that most immunolocalization I've seen is in TEM. I am } working in SEM. Is there any advantage of one over the other besides } the obvious difference that TEM looks inside the tissue and SEM looks } at the surface? How come there aren't as many SEM localizations out } there (that I've seen)? Thank you for your time, Karen } } ---------------------------------------------------------------------------
-- Regards, Gregory Rudomen Technical Specialist Electron Microscopy State University of New York at Stony Brook University Microscopy Imaging Center Stony Brook, NY 11794-8088 W-631-444-7372 Greg-at-umic.sunysb.edu--http://www.umic.sunysb.edu ************************************************* Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. *************************************************
The consensus clearly is Duniway Stockroom Corp. of Mountain View, CA. I called them and they have the proper cable in stock for less than half of what Varian wants. Your collective help is greatly appreciated.
If you first remove the plastic from your semi-thin sections, you can use most histological stains. A saturated solution of sodium hydroxide in methanol (sodium methoxide) will remove the plastic from 500 nm sections of plant material in about 10 minutes. Rinse the slides in 100% methanol followed by water and they are ready to go.
Tiny sections are sometimes very hard to find after the resin is removed, so we usually draw around the sections that have been heat-fixed to the slide with a glass scribe.
We (and others) have successfully used analine blue, Auramine O, and Sudan black on Arabidopsis. It is sometimes necessary to remove osmium from the de-plasticized sections (with sodium meta-periodate) for the staining protocols to work.
Most of these techniques are in M.A. Hayat's Principals and Techniques of Electron Microscopy Biological Applications.
If you'd like some references, I'll hunt them up.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Hi Ron, They are robbing you. For less than that price you should get a complete bellow including fittings and freight. That way you can continue operation with the old bellows silicone sealed until the replacment bellows are available. Disclaimer: ProSciTech supplies SS bellows. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, April 20, 2001 5:53 PM, Ron Doole [SMTP:ron.doole-at-materials.oxford.ac.uk] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Earl, } } We get the bellows replaced by a local (UK) firm, typically } around 250 pounds for edge welded bellows in stainless } steel. } This includes removing the old bellows, supply and welding } in new bellows and leak testing. It usually takes a few } weeks unless we are prepared to pay to interupt the work } scedule. } These prices are for 20-40mm dia 50-80mm long bellows. } } There must be firms in the US (and most other places) to do } this. } } Ron } } } On Thu, 19 Apr 2001 22:31:34 -0700 Earl Weltmer } {eweltmer-at-home.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi All, } } } } I have a Hitachi S-800 FESEM gun bellows that has developed a small leak. } } Although it has been replaced, I have heard of bellows being repaired by } } plating with cadmium or some other metal. } } } } Does anyone have any experience with this? } } } } Thank You, } } } } Earl Weltmer } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk }
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Karli Fitzelle writes: I have a quick question. I am working with purified nuclear matrix samples and need tips for the fixation / embedding process.......etc
Hi Karli, Chris Jeffree gave you the advice you need - embed the samples in low melting point agarose. Do this at the very beginning of the process after you have fixed them but washed away the fixative. You can then cut the embedded pieces into small blocks and do all your immunolabeling on these blocks. The agarose is permeable to antibodies and colloidal gold. You may need to increase your incubation and washing times to reflect the reduced accessibility. You will also not have to worry about centrifugation or rushing through the resin embedding as the sample will essentially behave as small tissue pieces.
If you do centrifuge the samples then do not worry about losing the gold - it will remain attached to the antibodies. Centrifuging the gold prior to labeling is another matter.
Regards,
Paul Webster
Paul Webster, Ph.D. Scientist II & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id KAA02466 for dist-Microscopy; Fri, 20 Apr 2001 10:21:44 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id KAA02463 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 20 Apr 2001 10:21:13 -0500 (CDT) Received: from hera.itg.uiuc.edu (hera.itg.uiuc.edu [130.126.126.145]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id KAA02456 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 20 Apr 2001 10:21:02 -0500 (CDT) Received: from buba.uiuc.edu (buba.itg.uiuc.edu [130.126.126.228]) by hera.itg.uiuc.edu (AIX4.3/8.9.3/8.9.3) with ESMTP id KAA257280 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 20 Apr 2001 10:17:51 -0500 Message-Id: {4.3.1.2.20010420085114.00bd5b40-at-mail.itg.uiuc.edu} X-Sender: gfried-at-mail.itg.uiuc.edu X-Mailer: QUALCOMM Windows Eudora Version 4.3.1
The Imaging Technology Group at the Beckman Institute, University of Illinois at Urbana-Champaign, is seeking a light microscopist to work in our multi-user Microscopy Suite.
Responsibilities will include:
· Operating/maintaining the confocal microscope, fluorescence microscope, stereology workstation, and stereo dissecting microscope. · Supervising and training others in the use of these instruments. · Working in conjunction with users to apply light microscopy techniques to their research. · Developing novel applications to take advantage of the unique capabilities of this instrumentation.
A complete job description is posted on the ITG web page http://www.itg.uiuc.edu or http://www.itg.uiuc.edu/ms/lightmicroscopistjob.pdf
Please send a letter of application and resume to:
Lori Heil Imaging Technology Group Beckman Institute for Advanced Science and Technology 405 N. Mathews Urbana, IL 61801 (217) 244-0170 e-mail: lheil-at-uiuc.edu
The University of Illinois is an Affirmative Action/Equal Opportunity Employer. Women and minorities are encouraged to apply.
Ray, I have made a number of specimen holders over the last few years, at rock- bottom prices :). My recipe for a pin-mount style mini-vice is: - grind or cut a small block of brass to the vice size desired - drill one hole in the large face of the block the size of the pin (typ. 3mm) - drill two parallel holes (the size for tap holes) through the long axis of the block - cut off one end of the block perpendicular to the two tap holes - tap the holes in the larger section of the block - drill out the tap holes in the smaller section of the block to make clearance holes - cut off a length of brass rod for the pin mount, and solder it in place in the hole in the larger block - screw two bolts (brass recommended) into the large block through the clearance holes in the smaller block. This is your mini-vice. - de-burr, clean, and polish as desired.
With proper tools several of these can be made in an hour; with what's available from the local hobby store, maybe 1.5-2 hours each.
Ben (simkin-at-egr.msu.edu)
} Howdy Y'all, } } I am looking for an inexpensive vise-style specimen holder for our } SEM. It does not need to be fancy, just a flat-jawed simple device } with a screw to tighten the faces of the jaws. After becoming } frustrated by all the information available on the web,I'm sure } someone out there could be of assistance. Please help by responding } online. } } Regards, } } Ray Grassl } } Grassl.raymond-at-basco.com
} } } } } } I have a user in our EM Center who would like to have a procedure for } } determining the thickness of their ultrathin sections, 60-100 nm. They are } } using the normal color criterion of "silver" sections in the boat to select } } their sections, but would like to be more precise. There is a potential } } for } } doing morphometry and comparisons of particle counts between non-serial } } sections and sections from different specimens. } } } } They have heard of a technique that uses small particles applied to both } } surfaces of the section and using tilt and geometry of the TEM stage to } } determine section thickness. Any details of this technique would be } } appreciated. } } } } All suggestions are welcome. } } } } John } } Colorado State University } } john.chandler-at-colostate.edu } }
The is a very good review of measuring section thickness in Audrey Glauert's book "Sectioning and Cryosectioning for Electron Microscopy". Several different methods are discussed. In short, it is difficult to know precisely how thick a silver section is. My edition of the book was published in 1991. There may be newer and more encouraging reports that I am not aware of.
Good Luck,
John
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
Karli, have you considered using magnetic beads to carry your organelles through these steps? The process is less disruptive to fragile organelles than centrifugation. Whether the gold will settle during centrifugation will depend on the size of the gold, an example is 15 nm. gold settles at 25,000 rpm for 60 minutes. For a discussion of this issue, you can consult the early papers describing the conjugation of various proteins to colloidal gold, Marge
Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation email: springett.margaret-at-mayo.edu
} ---------- } From: Karli Fitzelle } Sent: Thursday, April 19, 2001 2:27 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM Question
In a message dated 4/19/2001 3:01:22 AM Mountain Daylight Time, RCHIOVETTI-at-aol.com-at-sparc5.microscopy.com writes:
} Does anyone know of a digital imaging system for scanning EM?
A list of companies that provide digital imaging systems for SEMs can be found at "www.jcnabity.com/links.htm#Digital Imaging".
The two basic types of systems are passive (where they acquire an image while the SEM scans) and active (where they control the beam position during the image acquisition). The passive types will typically be less expensive, since they do not need to generate the sweep voltages. The active systems require that the SEM has XY inputs for beam control.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
I teach an undergraduate-level course in electron microscopy, and every year I find that the biggest hurdle for my students, not surprisingly, is the cutting of thin sections with glass knives. This is of course the point at which the students begin to get very discouraged with their individual research projects. I have tried various embedding media based on viscosity/penetration demands (Spurr's or ultra-low viscosity for plant tissue, for example), but I have been unable to come up with a medium that gives students a greater chance at success in cutting sections with glass knives. Does anyone have a favorite embedding medium that would allow fairly routine sectioning of diverse biological samples with glass knives. We are using primarily MT-2 microtomes.
Earl Weltmer wrote: ============================================= I have a Hitachi S-800 FESEM gun bellows that has developed a small leak. Although it has been replaced, I have heard of bellows being repaired by plating with cadmium or some other metal. ============================================== Is this one of those kinds of leaks that could be repaired, at least temporarily, with a product like VacSeal™ vacuum leak sealant? See URL http://www.2spi.com/catalog/vac/vacleak.html
It is used even on UHV systems for "temporary" repairs, but depending on the nature of the leak, sometimes these "temporary" repairs can last months or even years.
And this kind of fix is quick and cheap!
Disclaimer: SPI Supplies offers this product to those with vacuum leaks.
Chuck
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Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I'd like to prepare some TEM films of 30-50 Angstrom CdS particles that are in a block-co-polymer solution. Has anyone done this? Could we just pour them onto a carbon film and let it dry to examine the particles? Any other ideas?
I think that glass knives dull very rapidly (either damage or plastic build-up on the edge), and so we have always tried to get the first few sections at a given place on the knife edge, which tend to be the best. I wrote a brief procedure for students years ago, based on that approach, and have typed it below (the handout had a drawing of a glass knife, with area A = 1/3 of edge on the side where the whorl meets the edge, area B = middle 1/3 of knife edge, area C = 1/3 of edge at side where whorl is farthest from the edge):
1. Face the block in area C of knife (see diagram). Cut semithin sections (if desired) in area B of knife. Then move to area A.
2. Bring the block face parallel with the knife, using the shadow method. Then use the shadow to bring the block face as close to the knife (but without touching) as you dare.
3. With the ultramicrotome set for ultrathin sections, manually turn the microtome wheel quite rapidly until you see the first sign of contact (usually a sliver off one side of the block face), then stop turning.
4. Turn on automatic sectioning at usual slow cutting speed. Cut about 6-12 full-face sections, then stop and pick them up on EM grids.
5. Retract the stage slightly, move laterally to another place in area A, and repeat steps 2-4. Continue until area A has all been utilized (or until you have all the sections you need).
Good luck to you and your students.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Office: 5801 Medical Science II Building Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 E-mail: akc-at-umich.edu ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} I teach an undergraduate-level course in electron microscopy, and } every year I find that the biggest hurdle for my students, not } surprisingly, is the cutting of thin sections with glass knives. } This is of course the point at which the students begin to get very } discouraged with their individual research projects. I have tried } various embedding media based on viscosity/penetration demands } (Spurr's or ultra-low viscosity for plant tissue, for example), but I } have been unable to come up with a medium that gives students a } greater chance at success in cutting sections with glass knives. } Does anyone have a favorite embedding medium that would allow fairly } routine sectioning of diverse biological samples with glass knives. } We are using primarily MT-2 microtomes. } } I thank you in advance. } } Dick Briggs } Biology Department } Smith College
Karli, have you considered using magnetic beads to carry your organelles } through these steps? The process is less disruptive to fragile organelles } than centrifugation. Whether the gold will settle during centrifugation } will depend on the size of the gold, an example is 15 nm. gold settles at } 25,000 rpm for 60 minutes. For a discussion of this issue, you can consult } the early papers describing the conjugation of various proteins to colloidal } gold, } Marge
} Margaret Springett } IEM Specialist } Electron Microscopy Core Facility } Mayo Foundation } email: springett.margaret-at-mayo.edu
} } ---------- } } From: Karli Fitzelle } } Sent: Thursday, April 19, 2001 2:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: TEM Question
Robin, Why don't you try polymerizing the block co-polymer and then microtoming the sample. You could get someone over in the med school to do it for you. There are two places on campus that have the facilities and expertise to do it for you, biology, and pathology. The size of the particles would make it very straightforward to do. Talk to Jim Sheetz.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com } [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] } Sent: Friday, April 20, 2001 2:42 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Sample prep of CdS particles in block co-polymer mycells } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } } http://www.msa.microscopy.com/MicroscopyLists } erver/FAQ.html } } } } -------------------------------------------------------------- } ---------. } } } I'd like to prepare some TEM films of 30-50 Angstrom CdS } particles that are } in a block-co-polymer solution. Has anyone done this? Could } we just pour } them onto a carbon film and let it dry to examine the } particles? Any other } ideas? } } } Thanks, } } } Robin Griffin } UAB }
I worked in an MBE group where we grew epitaxial and heteroepitaxial layers on III-V compounds. I thought that I had a pretty good idea of what epitaxy is.
Recently, I have come across a rash of papers that claim epitaxial relationships across dissimilar materials with dissimilar crystal structures. This is just orientational relationships across the interface. It is not epitaxy! Is it because we as materials scientists/microscopists who know better are not getting to review these papers or has there been a change of the definition of epitaxy. What's going on? (This is a rhetorical question posed to foster a discussion.) What are we going to do about it? (another one.) Am I the only one that is concerned that the definition is becoming unclear?
I know that I have made comments about it when I review papers that claim epitaxy when it is really just an orientational relationship.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I am looking for a small oven in which to polymerize epoxy embedded samples (50-90 C). I'd like one small enough to fit into our fume cabinet, but the smallest one I can find is about 13 x 13 x15 inches. Does anyone know of a source for a smaller oven?
Thanks in advance,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
Does LKB still manufacture glass strips? If so, from whom does one obtain such glass? If not, what is the best glass available for sectioning and an approximate price? I vaguely remember "LKB glass" and have been using an unspecified "brand X". Thank you for your replies.
Barbara Plowman Univ. of the Pacific School of Dentistry 2155 Webster St. San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu ph: 415-929-6692
You might want to try contacting Stuart Enterprises run by Stuart Wisun. Specimen holders for SEMs are his specialty. His number is (650)424-9089, and his email address is stuwsn-at-juno.com.
Tom Pella
Raymond Grassl wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Howdy Y'all, } } I am looking for an inexpensive vise-style specimen holder for our } SEM. It does not need to be fancy, just a flat-jawed simple device } with a screw to tighten the faces of the jaws. After becoming } frustrated by all the information available on the web,I'm sure } someone out there could be of assistance. Please help by responding } online. } } Regards, } } Ray Grassl } } Grassl.raymond-at-basco.com
LKB is now part of Leica, but LKB never was a glass manufacturer. They marketed that glass only. Its been sold under the manufacturer's brand name "Alkar" for many years now. All EM suppliers sell that glass. We do sell Alkar glass in Australia and New Zealand, but clearly its one item that you would buy somewhere on your continent. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, April 21, 2001 8:40 AM, Barbara Plowman [SMTP:Bplowman-at-sfmail.dental.uop.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does LKB still manufacture glass strips? If so, from whom does one obtain } such glass? If not, what is the best glass available for sectioning and an } approximate price? I vaguely remember "LKB glass" and have been using an } unspecified "brand X". Thank you for your replies. } } Barbara Plowman } Univ. of the Pacific } School of Dentistry } 2155 Webster St. } San Francisco, CA 94115 } email: Bplowman-at-sf.uop.edu } ph: 415-929-6692 }
Robin Griffin wrote: ============================================================= I'd like to prepare some TEM films of 30-50 Angstrom CdS particles that are in a block-co-polymer solution. Has anyone done this? Could we just pour them onto a carbon film and let it dry to examine the particles? Any other ideas? ============================================================== What tends to happen is that the polymer is present in sufficient amount that it interferes with the imaging. Hence you can get around this problem by putting your preparation on silicon monoxide/dioxide filmed grids, then in an oxygen plasma etcher, etch away the polymer, leaving only the CdS colloid dispersed on the support film. However it might be better to put your solution on a glass slide, allow it to dry, and then etch off the organics as above, then Pt/C shadow or carbon coat only and pick up the replica on a grid and examine.
Disclaimer: SPI Supplies manufactures a plasma etcher and also produces SiO2 filmed grids.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Epitaxial growth is the oriented growth of a crystalline material over another crystalline material [9]. For example, films of fcc Au{001} can be grown by the deposition of gold on fcc Ag{001} surfaces; the silver lattice-parameter (4.08 Ang.) is very close to the gold lattice-parameter (4.09 Ang.). In some case the meshes may not match, e.g., Tb(0001) grown on W{110}, but incommensurate growth is still possible. In other cases, the meshes may match by the expansion, dilation, and rotation of the meshes, as Pb{111} grown on Si{111}. The success or failure of epitaxial growth depends highly upon the chosen material's chemical and physical properties, as well as the surface structure of the substrate.
} From: "Walck, Scott D." {walck-at-ppg.com} } To: "'Microscopy'" {microscopy-at-sparc5.microscopy.com} } Subject: Epitaxy -Am I misinformed or what? } Date: Fri, 20 Apr 2001 17:59:41 -0400 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I worked in an MBE group where we grew epitaxial and heteroepitaxial layers on III-V compounds. I thought that I had a pretty good idea of what epitaxy is. } } Recently, I have come across a rash of papers that claim epitaxial relationships across dissimilar materials with dissimilar crystal structures. This is just orientational relationships across the interface. It is not epitaxy! Is it because we as materials scientists/microscopists who know better are not getting to review these papers or has there been a change of the definition of epitaxy. What's going on? (This is a rhetorical question posed to foster a discussion.) What are we going to do about it? (another one.) Am I the only one that is concerned that the definition is becoming unclear? } } I know that I have made comments about it when I review papers that claim epitaxy when it is really just an orientational relationship. } } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } }
Since Joe's list for some reason unknown to me does not include my company, I would also like to invite you to take a look at our web site. We provide both passive and active systems.
Thanks.
Michael Bode Soft Imaging System Corp. www.soft-imaging.com mb-at-soft-imaging.com
-----Original Message----- } From: "NPGSlithography-at-aol.com"-at-sparc5.microscopy.com [mailto:"NPGSlithography-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, April 20, 2001 10:13 AM To: RCHIOVETTI-at-aol.com; Microscopy-at-sparc5.microscopy.com
In a message dated 4/19/2001 3:01:22 AM Mountain Daylight Time, RCHIOVETTI-at-aol.com-at-sparc5.microscopy.com writes:
} Does anyone know of a digital imaging system for scanning EM?
A list of companies that provide digital imaging systems for SEMs can be found at "www.jcnabity.com/links.htm#Digital Imaging".
The two basic types of systems are passive (where they acquire an image while the SEM scans) and active (where they control the beam position during the image acquisition). The passive types will typically be less expensive, since they do not need to generate the sweep voltages. The active systems require that the SEM has XY inputs for beam control.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Hi Dick Briggs: I was a former student at San Francisco State University (I've graduated in May 2000), and I took an electron microscopy course at that university. My instructor was Dr. Greg Antipa, and I'm sure that he can give you tips on plastics he used, etc. Back to my electron microscopy course ... during part of that course, I was supposed to section some embedded tissue of your choice (heart / liver / ? (can't recall off-hand)), and I had considerable trouble using a microtome well. As I recall, we had some Sorvalls available for use, but I'm unsure if they were MT-2s. I prepared all my tissues (and practice, plastic "pellets") by first cutting with straight razor blades to make an acceptable-sized pyramid, then continue by sectioning with a microtome. Glass knives were used on these embedded tissues -- the plastic used was Epon -- and I recall that one major problem I had was in making sure that the microtome sectioned at a consistent speed. In addition, the glass knife edges would dull pretty quickly after a small number of sections were made, and so it became imperative to use several glass knives for any particular embedded material. I eventually was able to section pretty well, but I also picked up sections from below with a grid (of 200 mesh) that I bent rather badly during the collection process. Epon worked OK for sectioning, and while it's true that students can indeed get discouraged, I also know from experience that students have very different aptitudes towards handling microtomes, preparing the pyramids, etc. I hope that my experience with sectioning may help you understand better (from a student's perspective) the various trouble areas that could occur when students are first learning how to section embedded material. Good luck with the sectioning process. Nelson Conti
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
On Monday April 23rd 2001 at ~ 9AM CST , after more than two decades of operation, the ANL HVEM will begin its last experimental session before permanently shutting down at the end of the day.
On behalf of the hundreds of users in the world wide microscopy research community who have used this facility since 1979, let me offer thanks to the ANL crew who has kept this unique resource alive, functional and running over this period: Hats off to ...Ed Ryan, Stan Ockers, Charlie Allen, Tony McCormick, as well as Alan Philippides, Loren Funk, Loren Thompson, Pete Baldo, and Tony Taylor.
If you would like to peer "live" into the microscope room via TelePresence and be a small part of the last day of operation of this unique instrument for Materials Research just point your Web browser to
http://tpm.amc.anl.gov/HVEM.html
When the last experiment completes later the afternoon of April 23rd, ANL will begin the decommissioning process and over the next few weeks the instrument will be completely dismantled. During this time we will endeavor to keep the live telepresence links operational to broadcast the decommissioning of this unique resource to allow interested students/researchers the opportunity to observe the process.
Coincidentally the last experiment being conducted on the HVEM will be a high energy electron irradiation damage study similiar in many respects to the first official experiment conducted at the facility during its opening ceremonies over twenty years ago.
Nestor J. Zaluzec Materials Science Division Argonne National Laboratory
-- =========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4289 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
Does anyone know how to detect glycolipid using LR-White/Cryo ultrathin-sections? I have tried both. But in cryosections the lipid is flowing out over the section, so when I do my immunogold, there is gold all over. I think that I lose some lipid bound to the membrane, when I use LR-White. Maybe because of the dehydrasion. Maybe there is a speciel way to fix the lipid?
The Biomaterials Science Group Department of Oral and Dental Science University of Bristol in collaboration with Glaxo SmithKline
Postdoctoral Research Assistant
in the Biomaterials Science Group on the project Interaction Mechanisms of Polymers at Interfaces of Mineralised Tissues
The research area involves the study of the physical and chemical properties and interaction mechanisms of different polymers at interfaces of mineralised tissues. You will have recently been awarded a PhD in an appropriate field and will ideally have experience in scanning probe microscopy (AFM) of biological materials and other analytical techniques and an interest in medical research. You will work in Dr. Jandts group and interact with scientists at Glaxo Smith Kline. The University of Bristol is one of the leading research universities in the UK and provides an outstanding scientific training environment to enhance your qualification. The group is involved in exciting, interdisciplinary projects and maintains appropriate state of the art instrumentation. There exist opportunities for additional interactions with clinical scientists and other centres at the university. We are looking for a dynamic and exceptionally well-qualified postdoctoral researcher who can interact effectively in an international and interdisciplinary team. The appointment will be on a Research Assistant 1A scale with a salary range of # 16775 to # 20465. This is a full time appointment and initially for one year. Applicants should include a short CV, stating research experience and interests, publication list and addresses of two referees. The review of applications will start 24 May 2001 and will continue until the post has been filled. Informal inquiries can be directed by email to Dr. K. D. Jandt (K.Jandt-at-bris.ac.uk), Senior Lecturer in Biomaterials, University of Bristol
Formal applications quoting the reference number 7401 should be directed to
The University of Bristol Recruitment Office Bristol, BS8 1TH United Kingdom
----------------------------------------------------------------- Dr. rer. nat. Klaus D. Jandt Senior Lecturer in Dental Materials Science and Biomaterials University of Bristol, Department of Oral and Dental Science Lower Maudlin Street, Bristol, BS1 2LY, UK Phone: +44-117-9284418, Fax: ++44-117-9284780 Internet: K.Jandt-at-bris.ac.uk WWW: http://www.dent.bris.ac.uk/Biomaterials/kdj.htm "We make Biomaterials Science work!"
Does anyone know how to detect glycolipid using LR-White/Cryo } ultrathin-sections? } I have tried both. But in cryosections the lipid is flowing out over the } section, so when I do my immunogold, there is gold all over. } I think that I lose some lipid bound to the membrane, when I use LR-White. } Maybe because of the dehydrasion. } Maybe there is a speciel way to fix the lipid?
What a sad morning this is. I've used this instrument, way back around 1980, and I can say it's a crying shame that such machines have such short lives. I found it to be quite useful for my area of study (finding the orientation distribution, more properly the misorientation distribution, in the cellular substructures of highly cold worked metals) for the simple reason that the selected area aperture of a high-voltage microscope can be demagnified sufficiently accurately for the electron beam to be made to illuminate the sub-micron-sized crystallites.
I've outlived _two_ high-voltage microscopes. Before the ANL instruments, I was a heavy user of the Million-Volt TEM at the US Steel Research Center, and for the same reason. Unfortunately, I was driven to use Argonne's HVEM because US Steel's hourly charges were too high to fit in my research budget; what with lack of support for what should have become a regional resource, the US Steel instrument was soon decommissioned just like ANL's is about to be.
Best regards, George Langford, Sc.D. amenex-at-amenex.com http://www.amenex.com/
Hi Scott, I think the term 'epitaxy' has indistinct boundaries. Okay, so InGaAs on GaAs is undisputedly epitaxy, as long as the crystals line up. But what about GaAs on Si? Most people would say this is epitaxy, but they're different crystal structures (diamond on sphalerite). Would you say something with rock salt structure can't be epitaxial with a diamond structure substrate? And just to stretch it a bit further, how about a (111) cubic layer on a (0001) hexagonal substrate? If not, what about hexagonal CdS on cubic CdS? My interpretation is that epitaxy requires both a reasonably fixed orientation relationship and a deposition. So MBE silicon on sapphire can be epitaxial, but a martensitic phase transformation can not. Maybe I should look it up in a science dictionary...
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I worked in an MBE group where we grew epitaxial and heteroepitaxial layers on III-V compounds. I thought that I had a pretty good idea of what epitaxy is. } } Recently, I have come across a rash of papers that claim epitaxial relationships across dissimilar materials with dissimilar crystal structures. This is just orientational relationships across the interface. It is not epitaxy! Is it because we as materials scientists/microscopists who know better are not getting to review these papers or has there been a change of the definition of epitaxy. What's going on? (This is a rhetorical question posed to foster a discussion.) What are we going to do about it? (another one.) Am I the only one that is concerned that the definition is becoming unclear? } } I know that I have made comments about it when I review papers that claim epitaxy when it is really just an orientational relationship. } } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } }
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
We are planning to do some 3D reconstruction of tick salivary glands from 2D light microscope sections (1 micron thick).
Does anyone have any suggestions for software (PC) that could do this.
thanks
Kevin
Kevin Mackenzie Electron Microscope unit Department Zoology University of Aberdeen Tillydrone Avenue Aberdeen AB24 2TZ ----------------- Tel 01224 272847 Fax 01224 272396 email k.s.mackenzie-at-abdn.ac.uk Web Site http://www.abdn.ac.uk/emunit
In June, I will have a mid-level EM Tech position open. One of my folks is getting married and moving away. We do all TEM--a mixture of negative staining and thin sectioning; 75% is clinical (diagnosing viral diseases with the EM, plus surgical pathology EM) and 25% is research. We also do some teaching of medical and graduate students and residents. We have 5 tech positions, and the 4 remaining individuals are all delightful folks with whom to work. Durham, NC, is a pleasant place in which to live and work--3 hrs from the mountains or the beach, and has more entertainment than you could want with 3 major universities and their arts programs in close proximity--not to mention the NCAA champs (Go Duke!).
Duke requirements for the position (EM Technician, Senior) include a bachelor¹s degree and electron microscopy training (a course or experience) or an associate¹s degree in EM. I am looking for someone who is proficient in cutting thin sections and has experience running an electron microscope. We can teach you to operate the particular scope brand we have, including digital operation for some applications, and we can teach negative staining and virus recognition. I am also looking for someone who enjoys challenging and interesting cases, is dedicated to accuracy, and is willing occasionally to put in extra effort in return for appropriate compensation and consideration when you have special needs. And I am particularly looking for someone who can manage several jobs at once while having a good time sharing camaraderie with the rest of us, i.e., is not high strung. I know this special person exists, since there are 4 lovely folks remaining (3 guys and 1 gal) with these same qualifications. I will be happy to answer any questions you have by phone or email.
If you might be interested, please contact me directly as the position is not officially open yet; I have requested that the paperwork be started. My address and phone number follow.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Hi, I need advice on embedding Thermanox coverslips. It's supposed to peel off leaving the monolayer behind, but I'm not having much luck. I'm using Aralidite 502 as the embedding medium. Would a another resin work better? I would appreciate any and all advice. Thanks.
Tom Bargar EM Lab UNMC 402-559-7347 tbargar-at-unmc.edu
A thread on plant cell wall staining for LM reminded me of a project wherein I would have liked to differentially stained parts of the plant cell wall - but for TEM. I could not readily find a good protocol for a CHO EM "stain" to do this. Does anyone have some good protocols and ideas for adding contrast or staining the plant cell wall?
I would like to express my heart felt sadness that another functioning HVEM will be decommissioned today. As one who has spent over 20 years working with almost the same microscope here in Albany, NY it is especially difficult to watch this days events. I know these microscopes like many know the inner workings of good vintage cars or the DC-3 and its difficult for me knowing how successfully the the ANL crew has been throughout the years with this very servicable machine.
There are 6 and then there 5 HVEMs in the US five years ago. Now there will be only 4!
I'm sure just like today we and NASA marvel at how quickly the technology and support for the Saturn 5 technology has disapeared even as our space program is just getting off low earth orbit, we in the microscopy field will find in the very near future how impossible it is to go back to the highvoltage technology that gave us that special edge for thick and dense specimens.
Dave
David Barnard HVEM Wadsworth Center NYS Dept Health Albany,NY
Autoquant's Autovizualize-3D will do this nicely. The resulting 3D image may be rotated and tilted and undergo further processing if desired.
Please contact me off-line if you are interested in this product or need assistance.
Gary Gaugler, Ph.D. Optical Reflections 916.791.8191 916.791.8186 7970 Twin Rocks Rd Granite Bay, CA 95746-8111 USA
Disclaimer: I am an authorized reseller of Autoquant's image processing software products. I mostly handle the Western US. If you have problems obtaining information on Autoquant products in the UK, please contact me for assistance.
See http://www.aqi.com for product technical info.
At 07:09 AM 4/23/2001, you wrote:
} Dear All } } We are planning to do some 3D reconstruction of tick salivary glands } from 2D light microscope sections (1 micron thick). } } Does anyone have any suggestions for software (PC) that could do this. } } thanks } } Kevin } } } Kevin Mackenzie } Electron Microscope unit } Department Zoology } University of Aberdeen } Tillydrone Avenue } Aberdeen } AB24 2TZ } ----------------- } Tel 01224 272847 } Fax 01224 272396 } email k.s.mackenzie-at-abdn.ac.uk } Web Site http://www.abdn.ac.uk/emunit
We still run a Kevex Quantum on a JEOL 840A SEM. We replaced the Delta V portion of our analyzer a few years ago, but still have the old chassis sitting here if you (or anyone else) ever need parts.
I believe most of these concepts were explained in either the Quantex manual (pg. G-65 for version 5)or in the Tutorial (pg.8-8) manual.
Working distance on an SEM was the distance from pole piece to sample surface. Fixed distance was the distance from the pole piece to the centerline of the detector crystal. Therefore, the combination of (WD-FD) and HD was used to calculate takeoff angle. Therefore, if you cannot get the exact numbers for your scope, you can probably measure the height difference between sample and detector and make up some half-reasonable numbers that generate the correct difference.
Azimuth angle fixed the detector position on the column. Assuming the sample holder tilts about the y-axis, azimuth was the angle between x-axis and the detector port. Thus, an azimuth angle of 0 degrees would mean that your sample tilts directly toward the detector. Our detector was located 23 degrees back of the x-axis. If you never tilt your sample, then azimuth would not matter. If you do, it helps to correct solid angle and takeoff angle for the effects of tilt.
We had a tilted detector on our 840. That made things a little trickier. We had a scale reading to show the distance from detector to sample. But since our detector was tilted, we could not use that straightaway for horizontal distance (HD). We had to look up FD and HD values from a table as a function of scale reading. It was a fairly simple exercise in geometry, but it always seemed strange that fixed distance (FD) was not really fixed when the detector was tilted.
I hope this gets you going. If not, just ask for clarification.
Warren
At 09:30 PM 4/19/2001 -0500, you wrote: } } Subject: EDXA, Need help with detector geometry } } } Sirs or Madams, } } } } } } I am running a JEOL CX II with a Kevex mod. 3200-0018 detector/ Kevex } Delta Class Anlyzer. } } } } } } I am having difficulty locating the geometric variables unique to this } } } mating of scope and detector. Kevex was unable to supply the data. } These } } } geometric variables are used by the analyzer software (Quantex) in } modeling and subtracting backgrounds. } } } } } } The variables I am unable to supply are Working Distance, Fixed Distance, } } } and Azimuth. I have seen reference to a Quantex Parameters List. This } } } document was shipped with the original equipment, but alas, this is a } } second hand scope and the detector was taken from the company warehouse. } } } } } } Does anybody use this combination of TEM and detector or know of someone } } } with this combination? Does anyone wish to share a document listing } } Quantex parameters for different scopes with Kevex detectors? } } } } } } My sanity is in your hands. } } } I remain humbly yours, } } } } } } Stephen Bennett } } } EMSL Analytical, Inc. } } } Miami, FL } } } } } } miamilab-at-emsl.com } } ---------------------- } Warren E. Straszheim } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking
Job Posting for Electron Bioimaging Lab, submitted by David P. Bazett-Jones
Service Manager, Electron Microscopy Facility
Date Posted: April 17, 2001
Position Status: Full-time, Fixed term
Department: Cell BiologyResearch Institute
Available: August 1, 2001
Description of the Position: You will share responsibility for the operation and maintenance of transmission and scanning electron microscopes in a new Bioimaging Facility co-sponsored by teaching hospitals in the University of Toronto. The microscopes include an ESEM (FEI/Philips) and a 200 kV TEM (FEI/Philips) equipped with EDX, GIF and cryostage. You will also be responsible for coordination and management
of electron bioimaging services required by investigators of the Hospital for Sick Children Research Institute. Qualifications: As an ideal candidate, you have completed a M.Sc. in biological sciences, or have completed a B.Sc. with experience in analytical electron microscopy, ultramicrotomy and other sample preparation techniques. Strong computer skills are an asset.You possess excellent verbal communication andorganizational skills. You have the ability to work well independently and in a team.
Hours : 35 hours/week
Salary: $39,848.95 - $50,277.67
Available to: Internal and External Candidates
Deadline: April 25, 2001
Submit Resume to : Erin O’HareThe Hospital for Sick Children, 555 University Avenue, Toronto, OntarioM5G1X8 Fax (416) 813-5671 E-mail: hr.recruiter-at-sickkids.on.ca
Must Quote File Number CG0102-EO We thank you in advance for your interest. Only those applicants selected for an interview will be contacted.
Hello Tom, I use Thermanox coverslips a lot here, so here is my advice: I always place the coverslip inverted on a drop of resin (if I don't, the coverslip will be "embedded" in the resin and is really hard to remove) on an aclar sheet. I have also found that the small weigh dishes, ours are white, are great for keeping many coverslips separate. Just put one coverslip per dish. They do tend to migrate on the aclar sheet, especially when the oven isn't level. Usually they peel off very easily. In the rare instance that they don't, I use a dissecting scope and choose the area I am interested it. I then use a razor blade and cut through the resin side and remove the area of interest. The small piece will come off nicely and is the exact size and shape I need to remount it on a "blank" block for sectioning. This is also great because the rest of the sample (cell
culture) remains whole and labelled for identification later. I hope this helps, Jo Dee
-- Jo Dee Fish Coordinator of Electron Microscopy Cell Analysis Facility The Burnham Institute 10901 N. Torrey Pines Rd. La Jolla, CA 92037 (858)646-3100 ext. 3620
Does anyone know if NaNO3 and KNO3 are or are not reasonably stable under an electron beam?
I want to use them as overlap standards for Na and K respectively, but would prefer to avoid explosions in the specimen chamber. They would be mounted on conductive double-sided tape, and carbon coated, so there is some oxidisable matter available.
Anybody been there?
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
To those biologist out there in cyber space who may have used monastral blue as a marker for macrophages years ago. Many years ago I purchased 3% monastral blue (copper phalocyanine) in solution from Sigma which we used for in vivo and in vitro phagocytic experiments. Unfortunately Sigma now only sell the powder form of monastral blue. Is there any one who has an old bottle of Sigma 3% monastral blue solution that can tell me what was the solvent used to prepare this solution so that I can dissolve this dye to obtain a 3% stock solution. I have tried every solution that I have in the laboratory to dissolve the powder without success. I need to know exactly what the solvent was in the old Sigma solution. Hope someone out there can help.
Terry Robertson (PhD)
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6009
The best way to clean immersion oil from a lens is to use lighter fluid. First remove as much oil from the lens with lens paper. Be gentle; don't rub. Then dip a cotton swab in the lighter fluid and lightly wipe it across the lens. All the oil will be removed and it will evaporate very quickly without leaving a residue or streaks. Xylene is not recommended as it can dissolve the adhesives holding the lens in place.
Cheryl Rehfeld Meyer Instruments, Inc. Leica Distributor -----Original Message----- } From: mckaylodge-at-aol.com {mckaylodge-at-aol.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Consider this option to an actual oven: I've used a TEMP-BLOK MODULE HEATER (by Lab-Line, distributed by Scientific Products) for curing resins. Its a small heater measuring about 5x8x3 inches, has high and low temperature ranges with variable control for each. They have a variety of removable blocks with arrays of wells in them into which you can put BEEM capsules, Eppendorf tubes, gelatin capsules,etc. I stick a thermometer in one of the wells to calibrate the tepmerature settings. Place a tight covering of aluminum foil over the top of the block to better stabilize the temperature inside.
I have bought them used from our University's scientific apparatus shop which trades in used lab gear.
Good luck,
Gib
} } Dear Microscopists, } } I am looking for a small oven in which to polymerize epoxy } embedded samples (50-90 C). I'd like one small enough to } fit into our fume cabinet, but the smallest one I can find } is about 13 x 13 x15 inches. Does anyone know of a source } for a smaller oven? } } Thanks in advance, } } Doug } ---------------------- } Douglas R. Keene } Associate Investigator } Shriners Hospital Research Facilities } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97201 } phone: 503-221-3434 } FAX: 503-412-6894 (9-5 PST) } e-mail: DRK-at-shcc.org } } } }
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
Consider this option to an actual oven: I've used a TEMP-BLOK MODULE HEATER (by Lab-Line, distributed by Scientific Products) for curing resins. Its a small electrical heating device measuring about 5x8x3 inches, has high and low temperature ranges with variable control for each. They have a variety of removable blocks with arrays of wells in them into which you can put BEEM capsules, Eppendorf tubes, gelatin capsules,etc. I stick a thermometer in one of the wells to calibrate the tepmerature settings. Place a tight covering of aluminum foil over the top of the block to better stabilize the temperature inside.
I have bought them used from our University's scientific apparatus shop which trades in used lab gear.
Good luck,
Gib
} } Dear Microscopists, } } I am looking for a small oven in which to polymerize epoxy } embedded samples (50-90 C). I'd like one small enough to } fit into our fume cabinet, but the smallest one I can find } is about 13 x 13 x15 inches. Does anyone know of a source } for a smaller oven? } } Thanks in advance, } } Doug} } ---------------------- } Douglas R. Keene } Associate Investigator } Shriners Hospital Research Facilities } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97201 } phone: 503-221-3434 } FAX: 503-412-6894 (9-5 PST) } e-mail: DRK-at-shcc.org -- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-1799 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
I would like to receive informations about seed tissues separation techniques to analyse under optical microscopy. Thanks for your attention Sinceraly Carla Bocchese
__________________________________________________________________________ Acesso fácil, rápido e ilimitado? Suporte 24hs? R$19,90? Só no AcessoBOL - http://www.bol.com.br/acessobol/
One day while walking down the street a highly successful HR Director was tragically hit by a bus and she died. Her soul arrived up in heaven where she was met at the Pearly Gates by St. Peter himself.
"Welcome to Heaven," said St. Peter. "Before you get settled in though, it seems we have a problem. You see, strangely enough, we've never once had a Human Resources Director make it this far and we're not really sure what to do with you."
"No problem, just let me in," said the woman.
"Well, I'd like to," replied St. Peter, "but I have higher orders. What we're going to do is let you have a day in Hell and a day in Heaven and then you can choose whichever one you want to spend an eternity in."
"Actually, I think I've made up my mind, I prefer to stay in Heaven", said the woman.
"Sorry, we have rules..." And with that St. Peter put the executive in an elevator and it went down-down-down to hell. The doors opened and she found herself stepping out onto the putting green of a beautiful golf course. In the distance was a country club and standing in front of her were all her friends - fellow executives that she had worked with and they were all dressed in evening gowns and cheering for her. They ran up and kissed her on both cheeks and they talked about old times.
They played an excellent round of golf and at night went to the country club where she enjoyed an excellent steak and lobster dinner. She met the Devil who was actually a really nice guy (kinda cute) and she had a great time telling jokes and dancing. She was having such a good time that before she knew it, it was time to leave. Everybody shook her hand and waved goodbye as she got on the elevator. The elevator went up-up-up and opened back up at the Pearly Gates and she found St. Peter waiting for her.
"Now it's time to spend a day in heaven," he said. So she spent the next 24 hours lounging around on clouds and playing the harp and singing. She had a great time and before she knew it her 24 hours were up and St Peter came and got her. "So, you've spent a day in hell and you've spent a day in heaven. Now you must choose your eternity," he said. The woman paused for a second and then replied, "Well, I never thought I'd say this, I mean, Heaven has been really great and all, but I think I had a better time in Hell." So St. Peter escorted her to the elevator and again she went down-down-down back to Hell.
When the doors of the elevator opened she found herself standing in a desolate wasteland covered in garbage and filth. She saw her friends were dressed in rags and were picking up the garbage and putting it in sacks. The Devil came up to her and put his arm around her.
"I don't understand," stammered the woman, "yesterday I was here and there was a golf course and a country club and we ate lobster and we danced and had a great time. Now all there is is a wasteland of garbage and all my friends look miserable."
The Devil looked at her and smiled. "Yesterday we were recruiting you, today you're staff..."
You may want to try dipping just the thermanox portion of your block into liquid nitrogen. The sudden change of temperature will likely loosen the thermanox away from your sample. I do not expect epon to be a problem, but I do know that it works well with Spurrs.
Good luck,
Doug
On Mon, 23 Apr 2001 10:12:10 -0500 "tbargar-at-unmc.edu"-at-sparc5.microscopy.com wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- Send Email } to ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } I need advice on embedding Thermanox coverslips. It's } supposed to peel off leaving the monolayer behind, but I'm } not having much luck. I'm using Aralidite 502 as the } embedding medium. Would a another resin work better? I } would appreciate any and all advice. Thanks. } } Tom Bargar } EM Lab } UNMC } 402-559-7347 } tbargar-at-unmc.edu } }
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
I need to find a commercial source for a calibrated search coil to check stray fields in microscope rooms. Does anyone have any experience with purchasing this item?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
You can buy calibrated digital magnetic field meters from F.W. Bell. The are good down to ~ 0.1 mGauss traceable to NIST. I have the Model 4080 which is a triaxial field measurement device.
http://www.fwbell.com
For Acoustic Measurements I use the EXTECH 407727 Digital Sound meter
http://www.extech.com
Nestor
Your Friendly Neighborhood SysOp
=============== } } I need to find a commercial source for a calibrated search coil to check } stray fields in microscope rooms. Does anyone have any experience with } purchasing this item? } } John C. Wheatley } Lab Manager } Arizona State University -- =========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4289 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
I need to find a commercial source for a calibrated search coil to check stray fields in microscope rooms. Does anyone have any experience with purchasing this item?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Hi John The best bet is to contact your local RS electronics supplier ( find them on http://www.rs-components.com/ )and ask for the ELF Magnetic Field Strength meter part number 212 837. We use it extensively for site tests and have found it to be just as accurate as using a professional test kit. The only difference between this unit and a search coil to a scope is that this device simply tells you if you have a field problem between 20 to 1200Hz where a more expensive search coil will tell you exactly what frequency it is that is causing the field.
Good Luck Luc Harmsen Anaspec, South Africa Technical support on microscopy. Tel + 27 (0) 11 476 3455 Fax + 27 (0) 11 476 7290 anaspec-at-icon.co.za www.anaspec.co.za
Remember, ICEM 15 will be in 2002, Durban, South Africa. www.icem15.com
-----Original Message----- } From: John C. Wheatley [mailto:John.Wheatley-at-asu.edu] Sent: 25 April 2001 01:10 To: Microscopy-at-sparc5.microscopy.com
I need to find a commercial source for a calibrated search coil to check stray fields in microscope rooms. Does anyone have any experience with purchasing this item?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Here, finally, are the responses I got to the query below:
A colleague has asked for recommendations for setting up a digital darkroom (fun to spend someone else's money!). This person would benefit from a really good scanner that could deal with prints, large format negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked into an Agfa Duoscan T2500. Do any of you have an opinion about this or other suitable scanners? ########## ********** We have the Duoscan 1200 but the 2500 is also a very nice unit--additional (real) resolution and a high O.D. range plus 14 or 16 bit image depth. The Agfa units also come with a built-in transparency plate (rather than having to add on a separate transparency adapter). I find that color fidelity is very good with the Duoscan, and Agfa provides both reflective and transparency calibration standards. I am currently scanning in 3x4 TEM negatives at 12 bits (yield is about 26MB per image), and scan time is fairly rapid. There is one option that you might want to consider: the DIImage unit is made for up to 4x5 negatives, and I think (can't remember the last time I read the specs--the neurons aren't firing today) that resolution is in the 2700 dpi range--even for the 4x5 size. ********** I have an Agfa duoscan, not the 2500 but a duoscan and I love it. I scan everything gels, ex-rays, 2x2,line art and em negatives. The scanner is very versatile. I would recommend it. ********** Got one, love it, wouldn't trade it for the world. OF course, we haven't had it long enough for little nitpicky things to start bugging me, but wow, does it do a nice job. We've scanned Polaroid Type 55 negatives, TEM negatives, TEM prints, Type 52 prints, and AFM 3-D presentations with excellent results. We have even scanned in old 35 mm slides (BW and color) and made prints from them, without enhancement, that are just as pleasing as the original slides. We don't regret the purchase. We were able to work through a local photography supply house and obtain a reconditioned one for a reasonable cost. Our biggest problem right now is finding a printer that will do it justice without mortgaging the farm. ********** } From a vendor: We have sold a large number of the Agfa T2500 into this market with great success.
The T2500 is an excellent scanner for TEM negs. The 2500x2500 optical resolution is high enough to capture fine details and still allow cropping or magnification of small areas of the negative. A 3.5 Dmax will capture details in the dense areas of negatives, slides or prints.
The Duoscan uses what Agfa calls "TwinPlate" design. Unlike most flatbed scanners which scan both reflective and film originals through the glass bed, the Duoscan holds films in a drawer, similar to a glassless negative carrier in an enlarger. No glass means no dust or scratches and no Newton Rings.
The Agfa Fotolook software driver is also excellent. Setting up scan parameters is easy and developing custom film terms is straightforward. You have control over almost every aspect of scanning, including a wide range of Gamma control which is beneficial for scanning very low or high contrast TEM negs.
For scanning 35mm slides, a batch holder accommodates up to 20 mounted 35mm slides. Holders are included for 35mm strips, mounted slides, 120/220 films, 4x5 films and a glass plate for odd size or other transparent originals. I like to scan TEM negs using the 4x5 holder, they fit across the opening and it works very well. ******************* I just bought an Epson Perfection 1240U for home use. I am impressed, So was a professional photographer friend - he was the lab photographer until made redundant recently.
It has a removable transmitted light box/lid combo, just lift off the normal lid from its extending hinge slots (to accommodate thickish documents/thin books?) and fit into the space on the flatbed. It has a separate light switch which normally cuts off when you finish a job - it seems.
It comes with a set of (thin, therefore, flimsy) plastic holders for 4 x 5, 120 roll film plus 35 mm and APS (a double strip holder). 35 mm slides have to be put directly on the flatbed.
It came with free Photoshop LE. "LE" meant no "channels" dialogue box! ******************** We have digital imaging equipment set up to produce photographic quality prints from transmission electron microscope negatives 6.5cm x 9.0cm, reproduce photograph prints for posters, prints from slides and of course convert all types of images suitable for email, to name but a few. The flat bed scanner is:- AGFA ARCUS II Slide scanner:- NIKON Coolscan II
Both have given good service. The quality produced by the ARCUS has not been fully exploited as we feel a compromise between file size and quality has to be a consideration. The software we use is Adobe Photoshop. Our equipment is certainly out of date now, but will be interested to hear what the new machines perform like. ********** There is a simple rule of thumb I use.
Nominal grain size of film is about 10 microns (varies with film speed etc but this is the right order of magnitude). Thus to digitize the film to it's nominal limits your scanner should be able to digitize to better than this spatial dimension.
A simple back of the envelope calculation says a spatial resolution of 10 microns is 2540 - dpi..... and as we all know that must be the optical resolution of the scanner not the interpolated resolution. Scanners at this end are obviously more than you need to digitize photo's and get expensive quickly. Also when you see 2 numbers listed as the scanners resolution, believe only the first number, that is the CCD resolution.
Now add your bit depth. 12 bits is the minimum I would shoot for grayscale image, but if your attempting diffraction work the higher the better (i.e. 14 -16 bits+). For color work obviously multiple the bit depth by 3 one for each primary color (RGB). I've seen a number of 36 bit color scanners but not too many 48 bit ones at } 2540 dpi.
Lastly, bit depth is irrelevant if you don't have a high optical density capabilities otherwise your just digitizing noise. The highest value I believe is an OD of 4.0 but this is for DRUM scanners. Flatbed scanners typically run as low as 2.8, upwards to about 3.4 for the best I've seen in a flatbed. ********** We have a Duoscan T2500, and I really like the resolution we can achieve when scanning any transparency media. There is no holder specifically designed for EM negs, but they fit sideways into the 4x5" holders. It's great for scanning Kodachromes; I was given a slide with a photo of someone (very small image) and was able to scan it in, cropped, at 4000 dpi, and turn that file into a 5x7" print without pixellation. Not bad.
The only drawback is that it can be painfully slow when calibrating. Still, I recommend it as a good, medium-to-high-end film scanner. It's also an excellent flatbed scanner, but with the low-end units available today, it's overkill. ********** Have your colleague check out the Imacon Flextight Precision II scanner. The optical resolution is 5760 dpi for slide-sized objects; I believe it drops to 4800 dpi for objects the size of her larger negatives. The scanner collects 14 bits of usable data per channel, which can be exported as a two bytes per channel, and has a dynamic range of 3.9 OD units (4.1 OD max). The machine is also very fast. The URL is: http://www.imacon.dk/usr/imacon/wppImacon.nsf/pages/flexprecision.html ********** If you are scanning EM negatives, you need to keep the dynamic range in mind. Regular flat beds are closer to 3.2 to 3.4 usually.
The ArtixScan 1100 has a Dmax of 3.9 (about $1600). This was has a 1000 x 2000 dpi resolution. more details at www.microtek.com
The Agfa DuoScan HiD (about $2400) has a 3.7 dynamic range. more details at www.agfa.com
Nikon has the new CoolScan 8000 that has a 4 or 4.2 dynamic range but it doesn't hold the large EM negative size - I think it is limited to something like 2.5 x 3.5 but their website http://www.klt.co.jp/Nikon/Press_Release/ls-8000_main.html has the details.
I think I am going to go with the ArtixScan and buy an extra template and have it machined to hold my size of negatives. Somewhere I saw scary data showing that it is important to support all 4 edges of the negative or you get significantly less optimal scans. The ArtixScan comes with 4 holders but none match my negative size exactly. It has a glass plate holder but the problem with these and any conventional flat bed scanner is that you get Newton rings on many or all of the scans if you look closely.
If you are willing to spend $14,000, there is a really neat film scanner called the Imacon Flextight Precision II CCD Drum Scanner that goes up to 5600 dpi (true optical) and 4.1 Dmax. I wish I could afford it. One web site with info about it is http://www.medgraphix.com/imaconscan.htm
a web site with really strong views on scanning negatives is http://www.flatbed-scanner-review.org/ ********** I have the HP Photosmart film scanner. It has a scanner of 2400 dpi, for 35 mm film. I think the recommendation of a film scanner is a good one for the following reason. Some scanner manufacturers make transparency (slide/negative) devices that use mirrors, but the image quality is poor. The Dimage or other large format scanners should provide acceptable images. The catch for large images and high resolution you need a lot of RAM memory. ********** I have the Duoscan and a Nikon slide scanner. The Duoscan can scan slides on the special tray feature but side by side comparisons of the Duoscan and Nikon show that the Nikon scan is much better. For the larger negs we had a special tray made for the Duoscan and we scan in our EM negs. The Nikon has gotten much cheaper and an excellent scanner can be had for $700 with Digital ICE, something you want. Get two scanners. ********** I love my Epson 1640 scanner, 1600x3200 and up to 4x5 negs and transparencies. ********** I was forwarded your inquiry into digital darkrooms by a colleague. I tackled this issue a few years back and the solution I arrived at is working out fine. I have been a professional photographer for 12 years. I work as an imaging specialist/photographer at a Materials Technology Laboratory. When our lab went digital (not yet 100%), I purchased what was then a very good flatbed scanner - Agfa Arcus II. It was a compromise of sorts. It could handle reflective and transparent originals. It has a max density of 3.2 and a max optical resolution of 1200 dpi. It is fine for in house publications and reports but falls short for anything going to a service bureau. I also don't recommend it for 35mm film. It can scan 35mm but not to the quality I required. We still use the Agfa for many scanning tasks but I have since purchased a more capable machine.
The new scanner is a Flextight Precision II, made by Imacon. It has a Dmax of 4.1, a true optical resolution of 5760 dpi and scans at 14 bits per colour. I purchased it primarily for it's density range. We have a large characterization section with a variety of beam instruments but the TEM negs were always tough to print. Some diffraction patterns take hours to print in a wet darkroom. I used the TEM negs as test samples for the scanners I was considering. A weak point of almost all the prospective film scanners was no holders for TEM film. Imacon has the capability of accepting custom made holders (Imacon will make them based on client specs). As well, the Precision II is primarily a film scanner. It will scan reflective originals up to A4, but I rarely use it for that.
If your colleague is looking for a flat bed scanner, Imacon makes a model called the Progression. It is equally as capable as the Precision but appears to handle reflective originals easier( it accepts film originals from 35mm to 5"x7").It also has a Dmax of 4.1, a true optical resolution of 5760 dpi and scans at 14 bits per colour. These are both quite a step up from the T2500. The 2500 boasts a resolution of 5000 dpi but that's interpolated resolution. I make it a practice not to interpolate when scanning scientific images because of the addition of false image information. The 2500 has a Dmax of 3.4 which is quite acceptable for correctly exposed film or originals with slight underexposure. I don't think it could handle a "Hail Mary" type of neg. With the Precision II, I've pulled quality information off a TEM neg in regions where it seemed transparent to the naked eye. I am very impressed with this machine. I don't want to seem indifferent to the T2500 however. I believe it is a good scanner and can handle most jobs with ease. I would also consider the acquisition software. Fotolook is quite good. I like it's tone curve editor. But Colorflex packaged with the Imacon scanners allows more manual control. It has Photoshop-like unsharp mask controls, good colour correction in all channels, ICC profiles, dot gain compensation etc.
I don't know your colleague's requirements. If he/she is looking for a capable, affordable desktop flatbed, I think you were quite correct to recommend the T2500. If he/she is hoping for more capability I would suggest they look into the Imacon line (www.imacon-usa.com).
The Imacon scanners are comparatively affordable. The Precision II is ~ $14,995 US and the Progression is ~ $19,995 US. I say comparatively because many comparable scanners are much more expensive ( priced between $14,000-$150,000). I realize it is a big jump from the $4500 from the T2500. I justified the expense with not only the quality increase but the time saved in the darkroom with trouble negatives. ********** I used Agfa DuoScan HiD earlier and I try to get it here as well. I like that machine a lot. It's optical resolution is 1000x2000 Dynamic range is 3.7D, which would help scanning DP's. If you want more info you can have a look at: http://www.agfa.com/scanners/duoscan_HiD.html Printing is another task you can buy things from AGFA as well. Their photoprinter is just excellent, but a bit expensive. I have tried nice HP inkjet printers with great success. ********** In response to Tina's post, I have not seen any mention on the list of the scanner I purchased a few weeks ago, the Epson Expression 1640XL. It has 1600dpi optical resolution (scans at a hardware resolution of 1600x3200 dpi) 42 bit color (14 bit gray) and Dmax of 3.6. It is large format, and the transparency adapter comes with a range of negative holders. Has SCSI or USB interfaces with firewire as an optional extra (I use USB on a Win 2000 system). Of course, you pay for what you get - it isn't cheap.
We are only just beginning to learn how best to use all the resolution and bit depth we now have, but I and my users love it!
This is not a comparison, of course (I haven't used the other models) but just to say we are happy with what we have. ********** We are getting first rate resolution results from our "UMAX Powerlock 1100 Magicscan" scanner coupled to a" FUJIX Pictography 3000" printer. Our base computer is always an Apple system upgraded periodically. ********** } In response to Tina's post, I have not seen any } mention on the list of the scanner I purchased } a few weeks ago, the Epson Expression 1640XL. } It has 1600dpi optical resolution (scans at } a hardware resolution of 1600x3200 dpi) 42 bit } color (14 bit gray) and Dmax of 3.6.
I would certainly believe the resolution and the color depth for this scanner is adequate, but if scanning TEM films is an issue, I'd seriously advise measuring the optical density of your films ... I've heard these approach OD} 4 ... which would imply you might consider the dedicated film scanners, e.g., Polaroid 45 Ultra or the new Nikon LS-8000. ********** I have the scanner you are looking at & like it a lot. To be quite honest I do not find that I need to exploit it's full capabilities. If I were in the market again, looking at newer technology I would be interested in a faster scanner of similar quality. Yes I want my cake & to eat it too :). I'll give you this analogy. If I have 10 negatives I will franchise my time, that is let things scan while I hang out in the office doing other things. If I have 20 negatives, I'll probably go to the darkroom to make photos. It is quicker & paper is cheaper. BTW I have an Epson 870 inkjet that produces nice quality images... cost is down to $180 US, (now the Epson 880)....no financial interest in these companies. ********** There was a thread recently on scanners for TEM film. I have looked up all the models mentioned, on the web and called agents for prices - and produced a comparative table, given below. I do not guarantee that the figures are accurate but they are my best interpretation of the data given. In the light of experience and Nestor's comments, I would suggest that 2000 dpi is a minimum for TEM negatives. You may be able to get away with less nine times out of ten, but there will be occasions when you need more. I would exclude the Minolta and all the Epsons from consideration (despite the incredibly low prices of some of the Epsons) because of the low pixel density. Among the rest the Nikon has the best pixel density and the best optical density (another critical parameter for TEM negatives). The price is very competitive too. The Nikon web site does not give a time for scanning a negative. On the face of it the Nikon would be a best buy - get a separate, inexpensive flatbed scanner for the other work. These comments are all my own opinions based on manufacturers' data. Since we are considering purchase any comments to the contrary would be most welcome. Code Maker Model Type A Agfa DuoScan T2500 Flatbed -Transparency included
B Epson 1640 several versions Flatbed -Transparency option 1680 several versions
C 1600 several versions Flatbed -Transparency included
D Imacon Flextight Precision II Drum -for film and large format
E Minolta Dimage ScanMulti II Film
F Nikon Super Coolscan 8000ED Film
G Polaroid 45 Ultra Film
H Umax Powerlook 3000 Flatbed -Transparency included
Code dpi OD Time Price Opinion at 6 x 9 cm
A 2500 x2500 3.4 3 min $4,500 Fair B 1600 x 3200 3.6 $300-$3000 Poor $800-$1400 Poor
C 1600 x 3200 3.3 $650-$1160 Not suitable
D 2240 x2240** 3.9/4.1 N/A above $10k Good: low pixel density
E 1128 x 1128 3.6 Not suitable
F 4000 x 4000 4.2 N/A $2,695 V. Good
G 2500 x 2500 3.8 5 min $7,495 Good but pricey
H 3048 x 3048 3.6 3 min $6,499
********** I too am about to buy and I would make a couple of comments on your evaluation. First, let me remind everyone that the Dynamic range is a log scale so small numerical differences are significant.
I also think the Nikon Coolscan 8000 looks great but it only takes a 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone to a smaller film size or is Nikon using a non-Japanese EM as their standard? seems odd but I don't see how the Nikon would be very useful. You say a {2000 line scanner would be useful 9 out of 10 times but want the 2000+ lines for the occasional high res scan. I would argue that the size of the negative was the more important variable to be worried about. The Nikon couldn't handle 4x5 LM negatives or transparencies from autoradiography of Westerns/Northerns, etc.
My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. more details at www.microtek.com. This is my leading candidate. It was 4 negative carriers and I await word whether one could be modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have my scientific instrumentation shop guys fabricate a holder. It comes with a glass 8 x 10 glass carrier for odd size negs but I want to avoid Newton rings and want a glassless carrier.
I would appreciate comments on the following argument (I think I have this correctly figured out but am not sure since so many out there seem to want to have a higher resolution scanner). I have a Fuji Pictrography 3000 printer with a 400 dpi output that is as good as any other widely available printer in the academic world. If you figure the maximum published image size is about 8 inches, that would mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% depending on the orientation of the negative and still be taking full advantage of the printer resolution. In reality, most EM publication prints are smaller than 8" wide so one could crop even more and still not need more than 1000 dpi. A resolution } 1000 dpi would be useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x 4 negative would be 192 MB. That is pretty big for doing morphometry on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB and that is much more manageable. Perhaps the difference is in the type of EM we are doing. I am working with biological specimens doing standard thin section type stuff. are you doing some Material Sci application that demands more? I would love to take advantage of the Firewire option but my information is that the 8700 has a Dmax of 3.4 vs the 3.9 for the 1100. That is a significant difference. Do EM negatives of biological thin sections reach that? I think so. I do a lot of EM immunocytochemistry and have to look for gold (intensely black) against a very dark tissue component so I am hoping the higher Dmax improves my results. I frequently scan negatives on a Umax 1100 (Dmax 3.4??) and can't differentiate the gold from the background although by eye I can discriminate them when the negative is placed on a light box. Changing my exposure would give me an unusable image for the rest of the tissue. Maybe this is an extreme case but I suspect that lots of "dark organelles" (e.g., lysosomes, nuclei) have fine structure that get lost in the scanning with a low Dmax scanner. ********** Your information is correct and mine is not. The Dmax of the 8700 is 3.4. ********** A colleague and I each recently bought Microtek scanners to scan TEM negatives. I have the Artixscan 1100 and he has the Model 8700 which has similar characteristics (actually higher resolution -1200dpi), 3.9 dmax at 42 bits color (14 grayscale), and the glassless film carrier setup. The 8700 has USB and Firewire interfaces and is cheaper ( {$1000), and the 1000 dpi Model 1100 has a SCSI interface. You might want to check out the specs of the lower cost model 8700 on the microtekusa website if your computer can handle USB or Firewire. Both scanners have performed up to our expectations, which I would characterize as modest. Microtek does not supply a 3-1/4 x 4 " negative carrier for standard size TEM film but you can easily make a serviceable one from stiff paper or light cardboard.
How much scanner resolution should you buy? The answer depends on how you intend to use it. Most applications do not require capturing the full resolution of the negative. From a practical viewpoint, the scanner resolution just determines how many times you can magnify the negative image to produce the final print size. For example, to get a publication-size print at 300 dpi, an image scanned at 1200 dpi scan could be zoomed 4X. A practical alternative to spending more for higher scanning resolution is to take photos at higher magnification. One exception is with lattice images from the TEM, which (depending on the lattice fringe spacing on the negative) might require higher scan resolutions to avoid getting a moire effect. (Of course, not everyone agrees. My colleague prefers to always scan at the maximum resolution).
What does a Dmax of 3.9 mean to you? To me it means a very dark negative. D is the log of the transmitted to incident intensity ratio. I wonder if users ever actually verify the manufacturer's specs with a calibrated density target. A Dmax of 3.9 can be useful for scanning TEM diffraction patterns that might have high contrast, but TEM micrograph negatives of metals and ceramics generally don't have that much contrast and biological thin section photos tend to have rather weak contrast. If your negatives are simply dark, use shorter photo exposure times. Scanning with maximum allowed grayscale resolutions (e.g., 14 bits rather than 8) is highly recommended if you intend to enhance or adjust images, but that's another story. I believe that those Agfa scanners are OEM by Microtek. If budget is the concern, I would recommend buying a Microtek Scanmaker 5 ($1,100) instead, which I have used for scanning quite a few EM negatives and have satisfactory results. The Dmax and the dynamic range for Scanmaker 5 is are about 3.7 and 3.4. Another model your colleague might consider is an Agfa Duoscan HiD ($ 2,500) which has a higher Dmax of 3.7, but less optical resolution of 1,000 dpi compared with 2500 dpi on a DuoScan T2500. What others failed to mentioned is that the DuoScan T2500 only has a narrow strip on the CCD bay being capable of scanning at 2,500 dpi, otherwise the true optical resolution is 1,000 dpi. Although a lot of investigators think that the higher scanning resolution, the better, my personal bias is leaning toward to purchase a scanner having at optical resolution at 1,000-1,200 dpi. Umax also carries a few mid- to high-end scanners such as Powerlook III for routine negative stains. My personal experience for the UMAX scanner is only limited to the Powerlook II, a mid-range scanner which gives more grayish scanned images compared to those high end models I mentioned previously. However, it is a descent scanner if you are working with color transparencies.
********** I have not summarized the lengthy thread about film (logarithmic) vs digital (linear) response!
Aloha, Tina
************************************************* * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} I am helping a former traditional photographic media user/computerphobe } set up digital imaging capabilities since her university/museum } department is closing their darkroom facilities and reassigning personnel } (sigh). She has what appears to be a decent budget (until I started } pricing the good stuff!). I am proposing she get a fast (733MHz) G4 Mac } with maximum (1.5GB) RAM, and she saw and fell in love with the Apple } 22" cinema display (as did I when I saw it in person). People seem to } like the Agfa T2500 scanner for prints and negatives. A moderate color } printer,since she has access to other really good printers in her department. A } Nikon Coolpix 990 digital camera for on- and off-microscope. Photoshop } 6.0, for which I'll train her. Corel Draw for vector graphics? ********** Adobe Illustrator or Freehand for vector graphics. Corel Draw is less common and has a proprietary file format that has caused me troubles preparing articles for Microscopy Today. Corel can save in other formats, but people have to use that option. Also, Adobe has educational pricing, and special package prices that bundle full versions of Photoshop and Illustrator.
The 733 Mhz G4 Mac is a great idea since it comes with the superdrive for making DVDs and CDs, which your friend will probably find very helpful for archiving. I would also encourage getting a copy of Retrospect for backups. I am a huge fan of HP printers and prefer them over Epson. I find the print quality as good or a bit better, and the software engines are much faster, in my experience with HP 970 cxi and Epson 750 and 850.
I also very much like the Nikon 990, although it has taken me a long time to figure out the best way to use it (almost too many options....especially for white balance and light metering, but the quality is very good). Make sure she gets a memory disk for the 990 that is large enough, 64 MB at least. If the memory card is too small she won't be able to save TIFF files, only JPG. She will need TIFF if anyone wants to do any sort of quantitative imaging, and the TIFF files are all well over 2 MB each. Get at least two cards and a USB card reader (these are cheap) so she can keep the card reader attached to the Mac, and just swap out memory cards instead of connecting the camera directly to the Mac, which is kind of a pain. Make sure she gets the AC adapter, too. Running the Nikon 990 off batteries is OK if you need to be mobile but in a lab setting swapping batteries gets old. Alternately, get a battery charger and extra NiCad batteries and keep the battery charger going all the time in a handy outlet. Then you avoid the extra AC power cord, which reduces cord tangle (a pet peeve of mine....). ********** If you want to do a darkroom for $10K you can do pretty well, especially if you don't need to include a killer printer. If you need a decent printer for proofing, etc, I would either use an inkjet or a low/mid end laser printer. Does she need color output from the local printer?
For the scanner, if she wanted one "great" scanner, the T2500 would be my choice, but here's a few things to consider. What makes a scanner great depends on the workflow.She should look at her workflow, in particular what amount of scanning from 35mm will be done? If there is much 35mm, will there be a large amount of 35mm slides where some type of auto feeder would be handy? Would scanning from uncut rolls of 35mm film be needed? The mix is the first thing to figure out. For occasional 35mm scanning the T2500 would do the job. As the mix of 35mm increases, the value of a dedicated 35mm scanner increases. One of the best new 35mm scanners will be the Nikon Supercoolscan 4000ED. 4000dpi res, great Dmax, plus a slide feeder, long roll holder and even an adapter for glass microscope slides are available.
On the TEM side, I think the T2500 is still the nuts, especially for cropping a smaller areas of a neg and still having the resolution to blow it up for larger prints.
If the scanner budget didn't allow for the T2500 or for the T2500 along with a 35mm scanner, the Duoscan Hi-D would be my next choice. The Hi-D has slightly lower res(2000x1000) but a high Dmax of 3.7. Agfa's software is also very powerful.
The first key is probably to figure how much she's going to spend on the workstation. After that do the printer and/ or the scanner(s).
Microtek has a new line called Artixscan. When you look at the specs you will notice many similarities to the Duoscans. This is because Microtek shares some of Agfa's hardware. Agfa makes their own apochromatic lenses and the software is different. I personally like Agfa's software better but Microtek does offer a slight price advantage. Attached is info on the Microtek 2500 and 1100 scanners. ************************* I both run the microscope labs here and am a researcher in materials engineering.
First I want to say that I strongly support the use of the digital laboratory. While we still occasionally use film for our highest quality requirements, in general we are fully digital. The use of digital cameras has really expanded our undergraduate teaching laboratories and has sped up our research.
I have found one "dark-side" to a digital imaging laboratory as a lab manager. As the lab manager, I have found that keeping a digital laboratory up-to-date is much more expensive than the film laboratory. When we were only film, we had to repair the film cartridges for our Polaroid PN film (it takes about five minutes) and have the microscopes cleaned about once a year at a cost of about $1k.
The digital lab. is much more expensive time and repair wise. Because we crunch our computers with our image size and storage, it takes more of my time to keep stuff going. All our computers are networked and in addition to work, the students tend to junk up the computers with downloads etc which stops them from working for the image processing work. This requires continual monitoring on my part (in spite of rules against using them for these applications!) In addition, keeping computers that will run the data is expensive. I buy pretty much the best out there, but somehow upgrades are still inevitable. I also have to supply print cartridges, etc. Researchers always supplied their own film and dark room supplies. In addition, I've had to have our cameras repaired numerous times. The cost was high (at least $500) and they stayed gone for up to a month. Finally, some of my cameras are about 3 years old. I can see a degradation in the image quality from when they were purchased. The cameras are much noisier. I see a future of regular replacement of my cameras in addition to the computer upgrades. So while the cost to the researchers is lower (which helps me as a researcher), the cost to the lab itself is higher (which hurts me as a lab manager). I'm working on setting up a fee schedule for this equipment but REALLY hate to have to do it. All of you who do this in a university know how painful it is!
Regarding the camera purchase-in addition to considering the camera resolution and cost, I think you should consider the image transfer. I recommend considering a camera with immediate transfer of the image to the microscope if you have numerous inexperienced users. Being able to focus on the screen is extremely helpful. The image transfer time is also important if you have many images to capture. We do image analysis on numerous images and some of the cameras have about a 30 second transfer time for decent resolution. This would be unbearable for the number of images we collect. I'm not sure how the Nikon Coolpix works but this should be considered by anyone that is purchasing a digital camera. ********** I just put together a nearly identical system: G4 533MHz, 1.5GB, 22" conventional monitor (but loved the cinema), Agfa and Polaroid 4x5 scanners, Nikon 990 and Fuji Pro S1, Photoshop, Adobe Illustrator, HP 5000PS poster printer and Adobe In-Design for laying out posters. Whew.....awesome.
Glad to see that you went with Macintosh. Very wise. Too many people fall for the empty promises made by the PC Windoze. ********** I agree that Corel Draw is less common, *especially for Macintosh*. If you were setting up a PC studio, I'd say otherwise, but Illustrator and Freehand have been the standard tools for Mac vector graphics for about a decade. I think Illustrator may have expanded portability with Photoshop since they're both made by Adobe. For the most part, Illustrator, Freehand and CorelDraw provide more or less the same features, they may just call their tools by different names. Once you know one, it's not too difficult to pick up the others for basic illustration. ********** I think you are on the right track. I think you might look at a dedicated slide scanner too. We got one years ago and it has seen lots of use. So many people have big collections of slides that they want to turn into digital images and the slide scanner fills the gap. It is quick, easy, and doesn't need much training to use. We also have a flat bed scanner, Arcus II, that gets lots of use, but I am glad we have the slide scanner too. Arranging slides and cropping etc, can be a pain on the flat bed. With the slide scanner, just slip in the slide and scan away. Ours is an old Polaroid Sprint Scan. Today you can get one for pretty cheap that is even better than ours.
I agree with the recommendation to go with Illustrator over Corel. Illustrator plays well with Photoshop and has never screwed us up. Sometimes we run into problems, Canvas has also been a trouble maker.
After a while we started having 'Disk Full' errors on the machine used with the scanners. Photoshop wants at least 2 - 3 times the size of the file on the 'Scratch Disk' to swap files etc. If the disk is getting full, you get stopped by no room on the 'Scratch Disk', same can happen when trying to print big files, need room to spool the file for printing. So, get some kind of removable medium, new Macs might have a CD-RW and that would be cool. I partitioned our big drive, setting aside 1 GB as a scratch disk where no files or other junk are allowed.
Slightly off the main topic, we have found a wide format printer gets lots of use. We have an HP 755CM. 36" wide color inkjet. People from all over use it to make posters for meetings and displays for classes etc.
I think you have started a very good discussion. It is one all of us are facing and things are changing so fast, we all need to benefit from the experience of each other. I don't mind at all that you 'introduced a subject that gets periodically posted here', its a new ballgame just about every six months. ********** } From last February(?)
} A broad question for the light microscopists- } } I'm writing up a wish list for our EM lab, and it includes (gasp) light } microscopes. My question is - how do I go about evaluating and choosing a } digital camera for light microscopes? It would be for both compound and } dissecting microscopes, should be color, decent resolution, not } necessarily low light nor real-time video, but capable of good images for } image analysis on sections. We are getting a confocal, so fluorescence } imaging would be done there rather than with the proposed 'scope and } camera. } } What do I need to look for, and what price ranges are we talking about? ********** Here is what I know from my explorations. We got a Kodak MDS 120 system. It is now obsolete and has been replace by a newer, higher resolution model. Cost was a couple of K. Half the cost was the adapter to get the camera on to the microscope.
Unlike many 35mm cameras, the lens of most digital cameras is not removable. So to get the camera to see what the microscope sees, there needs to be this special adapter gizmo. It is matched to the threads on the camera at one end and then to the microscope on the other end. The DC 120 is about 1K x 1K res. and does pretty well for up to 5" x 7" pictures. Even some 8" x 10" are OK. The key is to remember is a photo 'documentation' system. Good for reminding yourself of what you saw and good enough for most applications, but not the equal of film.
Using it is similar to film, only different. It works best in brightfield with plenty of light. It is rated at an equivalent of ASA 160. The overall appearance of the pictures looks harsher to me. The dynamic range seems to be more compressed than film. Adjusting the light for best contract is different than using film.
The biggest negative is the way it transfers the digital file from the camera to the computer. It goes something like this: Set up the camera like a film camera, fiddle around with the adapter (it is focusable) until the eyepieces and the camera are parfocal (not easy). If you are pretty close, take a preview picture. Wait. Wait. The picture has to be transferred from the camera to the computer. It is not real time like a video camera. After you wait, the image appears on the computer screen. If its OK, usually not, take a final high resolution picture. Wait longer while the larger file is transferred. Once its there, you are done. Ours uses a Photoshop plug in for acquisition so it easy going from here.
The trouble is that it is more like film in that you don't know if the picture is good the instant you expose it. Sometimes the light is not just right, sometimes the focus is a little off. You can't see these things in real time like you can with a video camera, but a video camera is really crummy resolution. If everything is set perfectly, then it is like exposing film, you take the picture and wait, a few seconds to a minute rather than a few days to a week to see the final result. I never worried about how the pictures would come out with film, they always did. But somehow not being able to see the final result instantly with the digital camera is very frustrating. Maybe since the digital camera is an add on it is not as well set up and out film camera for focus etc. I did find out that I had to get some non-adjustable eyepieces so the focus between the oculars and the camera would not change between users. We went round and round chasing focus until that was fixed.
This is another problem if you plan to use the camera on different microscopes. The adapter will have to be adjusted to the new system each time it is changed. A pain.
All that said, it has been useful having the camera. I learned a lot about digital photography, some people have gotten good use from it on a microscope, it is useful for regular photography, and I now can sound like an expert when discussing the pros and cons of systems with folks here and abroad who want to buy a system. ********** I looked at the MDS 290 also, Kodak was offering an 'upgrade' path from the 120. I passed because the 290's equiv. ASA is 100 vs 160 for the 120. Also the 290 would require USB for image transfer. It is supposed to be faster, but still won't be video rate. Also to get USB in the room with the microscope would require a new computer and since the old computer has a NuBus frame grabber for our video camera I would have to get a new frame grabber for the PCI bus new computer. So while the camera upgrade was pretty attractive, the total cost to upgrade was way more than could be justified by the amount of use that the digital camera has generated so far. I have heard a lot of interest in the Nikon Cool Pix 990 (I think that's the one). It apparently has the ability to send a lower res. B&W signal to a video monitor so focusing etc can be done in real time, then switch to high res color for the final digital shot. ******************************
Thanks to everyone who contributed!
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I wondered how commonly people use a tool (like a loop) to pick up ultrathin sections.
Does it make life easier or does it only introduce different problems?
I was told that I can buy some "luxury" - but am still undecided.
Your views would be very much appreciated.
Regards
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk Fax: 44(0)208 547 7562
Tina: Thanks for the time and effort you have invested in this. A most enlightening and useful summary.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc On Tue, 24 Apr 2001 21:58:02 -1000 (HST), Tina Carvalho wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | | Here, finally, are the responses I got to the query below: | | A colleague has asked for recommendations for setting up a digital | darkroom (fun to spend someone else's money!). This person would benefit | from a really good scanner that could deal with prints, large format | negatives (4"x5", 3.25"x4") as well as 35 mm slides. At one time I looked | into an Agfa Duoscan T2500. Do any of you have an opinion about this or | other suitable scanners? | ########## | ********** | We have the Duoscan 1200 but the 2500 is also a very nice unit--additional | (real) resolution and a high O.D. range plus 14 or 16 bit image | depth. The Agfa units also come with a built-in transparency plate | (rather than having to add on a separate transparency adapter). I find | that color fidelity is very good with the Duoscan, and Agfa provides both | reflective and transparency calibration standards. I am currently | scanning in 3x4 TEM negatives at 12 bits (yield is about 26MB per image), | and scan time is fairly rapid. There is one option that you might want to | consider: the DIImage unit is made for up to 4x5 negatives, and I think | (can't remember the last time I read the specs--the neurons aren't firing | today) that resolution is in the 2700 dpi range--even for the 4x5 size. | ********** | I have an Agfa duoscan, not the 2500 but a duoscan and I love it. I scan | everything gels, ex-rays, 2x2,line art and em negatives. The scanner is | very versatile. I would recommend it. | ********** | Got one, love it, wouldn't trade it for the world. OF course, we haven't | had it long enough for little nitpicky things to start bugging me, but | wow, does it do a nice job. We've scanned Polaroid Type 55 negatives, TEM | negatives, TEM prints, Type 52 prints, and AFM 3-D presentations with | excellent results. We have even scanned in old 35 mm slides (BW and | color) and made prints from them, without enhancement, that are just as | pleasing as the original slides. We don't regret the purchase. We were | able to work through a local photography supply house and obtain a | reconditioned one for a reasonable cost. Our biggest problem right now is | finding a printer that will do it justice without mortgaging the farm. | ********** | } From a vendor: We have sold a large number of the Agfa T2500 into this | market with great success. | | The T2500 is an excellent scanner for TEM negs. The 2500x2500 optical | resolution is high enough to capture fine details and still allow cropping | or magnification of small areas of the negative. A 3.5 Dmax will capture | details in the dense areas of negatives, slides or prints. | | The Duoscan uses what Agfa calls "TwinPlate" design. Unlike most flatbed | scanners which scan both reflective and film originals through the glass | bed, the Duoscan holds films in a drawer, similar to a glassless negative | carrier in an enlarger. No glass means no dust or scratches and no Newton | Rings. | | The Agfa Fotolook software driver is also excellent. Setting up scan | parameters is easy and developing custom film terms is | straightforward. You have control over almost every aspect of scanning, | including a wide range of Gamma control which is beneficial for scanning | very low or high contrast TEM negs. | | For scanning 35mm slides, a batch holder accommodates up to 20 mounted | 35mm slides. Holders are included for 35mm strips, mounted slides, | 120/220 films, 4x5 films and a glass plate for odd size or other | transparent originals. I like to scan TEM negs using the 4x5 holder, they | fit across the opening and it works very well. | ******************* | I just bought an Epson Perfection 1240U for home use. I am impressed, So | was a professional photographer friend - he was the lab photographer until | made redundant recently. | | It has a removable transmitted light box/lid combo, just lift off the | normal lid from its extending hinge slots (to accommodate thickish | documents/thin books?) and fit into the space on the flatbed. It has a | separate light switch which normally cuts off when you finish a job - it | seems. | | It comes with a set of (thin, therefore, flimsy) plastic holders for 4 x | 5, 120 roll film plus 35 mm and APS (a double strip holder). 35 mm slides | have to be put directly on the flatbed. | | It came with free Photoshop LE. "LE" meant no "channels" dialogue box! | ******************** | We have digital imaging equipment set up to produce photographic quality | prints from transmission electron microscope negatives 6.5cm x 9.0cm, | reproduce photograph prints for posters, prints from slides and of course | convert all types of images suitable for email, to name but a few. | The flat bed scanner is:- AGFA ARCUS II | Slide scanner:- NIKON Coolscan II | | Both have given good service. The quality produced by the ARCUS has not | been fully exploited as we feel a compromise between file size and quality | has to be a consideration. The software we use is Adobe Photoshop. | Our equipment is certainly out of date now, but will be interested to hear | what the new machines perform like. | ********** | There is a simple rule of thumb I use. | | Nominal grain size of film is about 10 microns (varies | with film speed etc but this is the right order of magnitude). | Thus to digitize the film to it's nominal limits your scanner should | be able to digitize to better than this spatial dimension. | | A simple back of the envelope calculation says a spatial resolution | of 10 microns is 2540 - dpi..... and as | we all know that must be the optical resolution of the | scanner not the interpolated resolution. Scanners at this | end are obviously more than you need to digitize photo's and | get expensive quickly. Also when you see 2 numbers listed | as the scanners resolution, believe only the first number, that is | the CCD resolution. | | Now add your bit depth. 12 bits is the minimum I | would shoot for grayscale image, but if your attempting | diffraction work the higher the better (i.e. 14 -16 bits+). | For color work obviously multiple the bit depth by 3 | one for each primary color (RGB). I've seen a number | of 36 bit color scanners but not too many 48 bit ones at | } 2540 dpi. | | Lastly, bit depth is irrelevant if you don't have a high | optical density capabilities otherwise your just digitizing | noise. The highest value I believe is an OD of 4.0 but | this is for DRUM scanners. Flatbed scanners typically | run as low as 2.8, upwards to about 3.4 for the best | I've seen in a flatbed. | ********** | We have a Duoscan T2500, and I really like the resolution we can achieve | when scanning any transparency media. There is no holder specifically | designed for EM negs, but they fit sideways into the 4x5" holders. It's | great for scanning Kodachromes; I was given a slide with a photo of | someone (very small image) and was able to scan it in, cropped, at 4000 | dpi, and turn that file into a 5x7" print without pixellation. Not bad. | | The only drawback is that it can be painfully slow when calibrating. | Still, I recommend it as a good, medium-to-high-end film scanner. It's | also an excellent flatbed scanner, but with the low-end units available | today, it's overkill. | ********** | Have your colleague check out the Imacon Flextight Precision II | scanner. The optical resolution is 5760 dpi for slide-sized objects; | I believe it drops to 4800 dpi for objects the size of her larger | negatives. The scanner collects 14 bits of usable data per channel, | which can be exported as a two bytes per channel, and has a dynamic | range of 3.9 OD units (4.1 OD max). The machine is also very fast. | The URL is: | http://www.imacon.dk/usr/imacon/wppImacon.nsf/pages/flexprecision.html | ********** | If you are scanning EM negatives, you need to keep the dynamic range in | mind. Regular flat beds are closer to 3.2 to 3.4 usually. | | The ArtixScan 1100 has a Dmax of 3.9 (about $1600). This was has a 1000 x | 2000 dpi resolution. more details at www.microtek.com | | The Agfa DuoScan HiD (about $2400) has a 3.7 dynamic range. more details | at www.agfa.com | | Nikon has the new CoolScan 8000 that has a 4 or 4.2 dynamic range but it | doesn't hold the large EM negative size - I think it is limited to | something like 2.5 x 3.5 but their website | http://www.klt.co.jp/Nikon/Press_Release/ls-8000_main.html has the | details. | | I think I am going to go with the ArtixScan and buy an extra template and | have it machined to hold my size of negatives. Somewhere I saw scary data | showing that it is important to support all 4 edges of the negative or you | get significantly less optimal scans. The ArtixScan comes with 4 holders | but none match my negative size exactly. It has a glass plate holder but | the problem with these and any conventional flat bed scanner is that you | get Newton rings on many or all of the scans if you look closely. | | If you are willing to spend $14,000, there is a really neat film scanner | called the Imacon Flextight Precision II CCD Drum Scanner that goes | up to 5600 dpi (true optical) and 4.1 Dmax. I wish I could afford it. | One web site with info about it is | http://www.medgraphix.com/imaconscan.htm | | a web site with really strong views on scanning negatives is | http://www.flatbed-scanner-review.org/ | ********** | I have the HP Photosmart film scanner. It has a scanner of 2400 dpi, for | 35 mm film. I think the recommendation of a film scanner is a good one | for the following reason. Some scanner manufacturers make transparency | (slide/negative) devices that use mirrors, but the image quality is poor. | The Dimage or other large format scanners should provide acceptable | images. The catch for large images and high resolution you need a lot of | RAM memory. | ********** | I have the Duoscan and a Nikon slide scanner. The Duoscan can scan slides | on the special tray feature but side by side comparisons of the Duoscan | and Nikon show that the Nikon scan is much better. For the larger negs we | had a special tray made for the Duoscan and we scan in our EM negs. The | Nikon has gotten much cheaper and an excellent scanner can be had for $700 | with Digital ICE, something you want. Get two scanners. | ********** | I love my Epson 1640 scanner, 1600x3200 and up to 4x5 negs and | transparencies. | ********** | I was forwarded your inquiry into digital darkrooms by a colleague. I | tackled this issue a few years back and the solution I arrived at is | working out fine. I have been a professional photographer for 12 years. I | work as an imaging specialist/photographer at a Materials Technology | Laboratory. | When our lab went digital (not yet 100%), I purchased what was then a | very good flatbed scanner - Agfa Arcus II. It was a compromise of | sorts. It could handle reflective and transparent originals. It has a max | density of 3.2 and a max optical resolution of 1200 dpi. It is fine for in | house publications and reports but falls short for anything going to a | service bureau. I also don't recommend it for 35mm film. It can scan 35mm | but not to the quality I required. We still use the Agfa for many scanning | tasks but I have since purchased a more capable machine. | | The new scanner is a Flextight Precision II, made by Imacon. It has a Dmax | of 4.1, a true optical resolution of 5760 dpi and scans at 14 bits per | colour. I purchased it primarily for it's density range. We have a large | characterization section with a variety of beam instruments but the TEM | negs were always tough to print. Some diffraction patterns take hours to | print in a wet darkroom. I used the TEM negs as test samples for the | scanners I was considering. A weak point of almost all the prospective | film scanners was no holders for TEM film. Imacon has the capability of | accepting custom made holders (Imacon will make them based on client | specs). As well, the Precision II is primarily a film scanner. It will | scan reflective originals up to A4, but I rarely use it for that. | | If your colleague is looking for a flat bed scanner, Imacon makes a model | called the Progression. It is equally as capable as the Precision but | appears to handle reflective originals easier( it accepts film originals | from 35mm to 5"x7").It also has a Dmax of 4.1, a true optical resolution | of 5760 dpi and scans at 14 bits per colour. These are both quite a step | up from the T2500. The 2500 boasts a resolution of 5000 dpi but that's | interpolated resolution. I make it a practice not to interpolate when | scanning scientific images because of the addition of false image | information. The 2500 has a Dmax of 3.4 which is quite acceptable for | correctly exposed film or originals with slight underexposure. I don't | think it could handle a "Hail Mary" type of neg. With the Precision II, | I've pulled quality information off a TEM neg in regions where it seemed | transparent to the naked eye. I am very impressed with this machine. I | don't want to seem indifferent to the T2500 however. I believe it is a | good scanner and can handle most jobs with ease. I would also consider the | acquisition software. Fotolook is quite good. I like it's tone curve | editor. But Colorflex packaged with the Imacon scanners allows more manual | control. It has Photoshop-like unsharp mask controls, good colour | correction in all channels, ICC profiles, dot gain compensation etc. | | I don't know your colleague's requirements. If he/she is looking for a | capable, affordable desktop flatbed, I think you were quite correct to | recommend the T2500. If he/she is hoping for more capability I would | suggest they look into the Imacon line (www.imacon-usa.com). | | The Imacon scanners are comparatively affordable. The Precision II is ~ | $14,995 US and the Progression is ~ $19,995 US. I say comparatively | because many comparable scanners are much more expensive ( priced between | $14,000-$150,000). I realize it is a big jump from the $4500 from the T2500. I | justified the expense with not only the quality increase but the time | saved in the darkroom with trouble negatives. | ********** | I used Agfa DuoScan HiD earlier and I try to get it here as well. I like | that machine a lot. It's optical resolution is 1000x2000 Dynamic range is | 3.7D, which would help scanning DP's. If you want more info you can have a | look at: | http://www.agfa.com/scanners/duoscan_HiD.html | Printing is another task you can buy things from AGFA as well. Their | photoprinter is just excellent, but a bit expensive. I have tried nice HP | inkjet printers with great success. | ********** | In response to Tina's post, I have not seen any mention on the list of the | scanner I purchased a few weeks ago, the Epson Expression 1640XL. It has | 1600dpi optical resolution (scans at a hardware resolution of 1600x3200 | dpi) 42 bit color (14 bit gray) and Dmax of 3.6. It is large format, and | the transparency adapter comes with a range of negative holders. Has SCSI | or USB interfaces with firewire as an optional extra (I use USB on a Win | 2000 system). Of course, you pay for what you get - it isn't cheap. | | We are only just beginning to learn how best to use all the resolution and | bit depth we now have, but I and my users love it! | | This is not a comparison, of course (I haven't used the other models) but | just to say we are happy with what we have. | ********** | We are getting first rate resolution results from our "UMAX Powerlock | 1100 Magicscan" scanner coupled to a" FUJIX Pictography | 3000" printer. Our base computer is always an Apple system upgraded | periodically. | ********** | } In response to Tina's post, I have not seen any | } mention on the list of the scanner I purchased | } a few weeks ago, the Epson Expression 1640XL. | } It has 1600dpi optical resolution (scans at | } a hardware resolution of 1600x3200 dpi) 42 bit | } color (14 bit gray) and Dmax of 3.6. | | I would certainly believe the resolution and the color depth for | this scanner is adequate, but if scanning TEM films is an issue, I'd | seriously advise measuring the optical density of your films ... I've | heard these approach OD} 4 ... which would imply you might consider the | dedicated film scanners, e.g., Polaroid 45 Ultra or the new Nikon | LS-8000. | ********** | I have the scanner you are looking at & like it a lot. To be quite honest | I do not find that I need to exploit it's full capabilities. If I were in | the market again, looking at newer technology I would be interested in a | faster scanner of similar quality. Yes I want my cake & to eat it too :). | I'll give you this analogy. If I have 10 negatives I will franchise my | time, that is let things scan while I hang out in the office doing other | things. If I have 20 negatives, I'll probably go to the darkroom to make | photos. It is quicker & paper is cheaper. BTW I have an Epson 870 inkjet | that produces nice quality images... cost is down to $180 US, (now the | Epson 880)....no financial interest in these companies. | ********** | There was a thread recently on scanners for TEM film. I have looked up | all the models mentioned, on the web and called agents for prices - and | produced a comparative table, given below. | I do not guarantee that the figures are accurate but they are my best | interpretation of the data given. | In the light of experience and Nestor's comments, I would suggest that | 2000 dpi is a minimum for TEM negatives. You may be able to get away | with less nine times out of ten, but there will be occasions when you | need more. | I would exclude the Minolta and all the Epsons from consideration | (despite the incredibly low prices of some of the Epsons) because of the | low pixel density. | Among the rest the Nikon has the best pixel density and the best optical | density (another critical parameter for TEM negatives). The price is | very competitive too. The Nikon web site does not give a time for | scanning a negative. On the face of it the Nikon would be a best buy - | get a separate, inexpensive flatbed scanner for the other work. | These comments are all my own opinions based on manufacturers' data. | Since we are considering purchase any comments to the contrary would be | most welcome. | Code Maker Model Type | A Agfa DuoScan T2500 Flatbed | -Transparency | included | | B Epson 1640 several versions Flatbed | -Transparency | option | 1680 several versions | | C 1600 several versions Flatbed | -Transparency | included | | D Imacon Flextight Precision II Drum -for film and | large | format | | E Minolta Dimage ScanMulti II Film | | F Nikon Super Coolscan 8000ED Film | | G Polaroid 45 Ultra Film | | H Umax Powerlook 3000 Flatbed | -Transparency | included | | | | | | Code dpi OD Time Price | Opinion | at 6 x 9 cm | | | A 2500 x2500 3.4 3 min $4,500 | Fair | B 1600 x 3200 3.6 $300-$3000 | Poor | $800-$1400 | Poor | | C 1600 x 3200 3.3 $650-$1160 | Not suitable | | D 2240 x2240** 3.9/4.1 N/A above $10k | Good: low pixel density | | E 1128 x 1128 3.6 | Not suitable | | F 4000 x 4000 4.2 N/A $2,695 | V. Good | | G 2500 x 2500 3.8 5 min $7,495 | Good but pricey | | H 3048 x 3048 3.6 3 min $6,499 | | ********** | I too am about to buy and I would make a couple of comments on your | evaluation. First, let me remind everyone that the Dynamic range is | a log scale so small numerical differences are significant. | | I also think the Nikon Coolscan 8000 looks great but it only takes a | 2.5 x 3.5 negative which is smaller than my JEOL and Hitachi EM | negative sizes (~ 3 1/2 by 4 1/2"). Have these EM manufacturers gone | to a smaller film size or is Nikon using a non-Japanese EM as their | standard? seems odd but I don't see how the Nikon would be very | useful. You say a {2000 line scanner would be useful 9 out of 10 | times but want the 2000+ lines for the occasional high res scan. I | would argue that the size of the negative was the more important | variable to be worried about. The Nikon couldn't handle 4x5 LM | negatives or transparencies from autoradiography of | Westerns/Northerns, etc. | | My leading candidate is the ArtixScan 1100 has a Dmax of 3.9 (about | $1600 with SCSI card). This was has a 1000 x 2000 dpi resolution. | more details at www.microtek.com. This is my leading candidate. It | was 4 negative carriers and I await word whether one could be | modified to carry a 3 1/2 by 4 1/2 negative. At worst, I will have | my scientific instrumentation shop guys fabricate a holder. It comes | with a glass 8 x 10 glass carrier for odd size negs but I want to | avoid Newton rings and want a glassless carrier. | | I would appreciate comments on the following argument (I think I have | this correctly figured out but am not sure since so many out there | seem to want to have a higher resolution scanner). I have a Fuji | Pictrography 3000 printer with a 400 dpi output that is as good as | any other widely available printer in the academic world. If you | figure the maximum published image size is about 8 inches, that would | mean the maximum image size be 3200 dpi wide. A 1000 dpi scan of my | negative would be 4500 x 3500 dpi. I could crop by about 28% or 10% | depending on the orientation of the negative and still be taking full | advantage of the printer resolution. In reality, most EM publication | prints are smaller than 8" wide so one could crop even more and still | not need more than 1000 dpi. A resolution } 1000 dpi would be | useful for subtle morphometric analysis but a 4000 dpi scan of a 3 x | 4 negative would be 192 MB. That is pretty big for doing morphometry | on! A 1000 dpi scan of a 3.5 x 4.5" negative would be about 16 MB | and that is much more manageable. Perhaps the difference is in the | type of EM we are doing. I am working with biological specimens | doing standard thin section type stuff. are you doing some Material | Sci application that demands more? | I would love to take advantage of the Firewire option but my | information is that the 8700 has a Dmax of 3.4 vs the 3.9 for the | 1100. That is a significant difference. Do EM negatives of | biological thin sections reach that? I think so. I do a lot of EM | immunocytochemistry and have to look for gold (intensely black) | against a very dark tissue component so I am hoping the higher Dmax | improves my results. I frequently scan negatives on a Umax 1100 | (Dmax 3.4??) and can't differentiate the gold from the background | although by eye I can discriminate them when the negative is placed | on a light box. Changing my exposure would give me an unusable | image for the rest of the tissue. Maybe this is an extreme case but | I suspect that lots of "dark organelles" (e.g., lysosomes, nuclei) | have fine structure that get lost in the scanning with a low Dmax | scanner. | ********** | Your information is correct and mine is not. The Dmax of the 8700 is 3.4. | ********** | A colleague and I each recently bought Microtek scanners to scan TEM | negatives. I have the Artixscan 1100 and he has the Model 8700 which has | similar characteristics (actually higher resolution -1200dpi), 3.9 dmax at | 42 bits color (14 grayscale), and the glassless film carrier setup. The | 8700 has USB and Firewire interfaces and is cheaper ( {$1000), and the 1000 | dpi Model 1100 has a SCSI interface. You might want to check out the | specs of the lower cost model 8700 on the microtekusa website if your | computer can handle USB or Firewire. | Both scanners have performed up to our expectations, which I would | characterize as modest. Microtek does not supply a 3-1/4 x 4 " negative | carrier for standard size TEM film but you can easily make a serviceable | one from stiff paper or light cardboard. | | How much scanner resolution should you buy? The answer depends on how you | intend to use it. Most applications do not require capturing the full | resolution of the negative. From a practical viewpoint, the scanner | resolution just determines how many times you can magnify the negative | image to produce the final print size. For example, to get a | publication-size print at 300 dpi, an image scanned at 1200 dpi scan could | be zoomed 4X. A practical alternative to spending more for higher | scanning resolution is to take photos at higher | magnification. One exception is with lattice images from the TEM, which | (depending on the lattice fringe spacing on the negative) might require | higher scan resolutions to avoid getting a moire effect. (Of course, not | everyone agrees. My colleague prefers to always scan at the maximum | resolution). | | What does a Dmax of 3.9 mean to you? To me it means a very dark | negative. D is the log of the transmitted to incident intensity ratio. I | wonder if users ever actually verify the manufacturer's specs with a | calibrated density target. A Dmax of 3.9 can be useful for scanning TEM | diffraction patterns that might have high contrast, but TEM micrograph | negatives of metals and ceramics generally don't have that much contrast | and biological thin section photos tend to have rather weak contrast. If | your negatives are simply dark, use shorter photo exposure | times. Scanning with maximum allowed grayscale resolutions | (e.g., 14 bits rather than 8) is highly recommended if you intend to | enhance or adjust images, but that's another story. | I believe that those Agfa scanners are OEM by Microtek. If budget is the | concern, I would recommend buying a Microtek Scanmaker 5 ($1,100) instead, | which I have used for scanning quite a few EM negatives and have | satisfactory results. The Dmax and the dynamic range for Scanmaker 5 is | are about 3.7 and 3.4. Another model your colleague might consider is an | Agfa Duoscan HiD ($ 2,500) which has a higher Dmax of 3.7, but less | optical resolution of 1,000 dpi compared with 2500 dpi on a DuoScan | T2500. What others failed to mentioned is that the DuoScan T2500 only has | a narrow strip on the CCD bay being capable of scanning at 2,500 dpi, | otherwise the true optical resolution is 1,000 dpi. Although a lot of | investigators think that the higher scanning resolution, the better, my | personal bias is leaning toward to purchase a scanner having at optical | resolution at 1,000-1,200 dpi. Umax also carries a few mid- to high-end scanners such as | Powerlook III for routine negative stains. My personal experience for the | UMAX scanner is only limited to the Powerlook II, a mid-range scanner | which gives more grayish scanned images compared to those high end models | I mentioned previously. However, it is a descent scanner if you are | working with color transparencies. | | ********** | I have not summarized the lengthy thread about film (logarithmic) vs | digital (linear) response! | | | Aloha, | Tina | | ************************************************* | * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * | * Biological Electron Microscope Facility * (808) 956-6251 * | * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* | **************************************************************************** | | | | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Question: I am trying to do immunohistochemistry on lung tissue. What is the best way to fix this tissue? Is there a good reference for protocols? Thank you
A slip of the finger caused me to send an un-topical joke to all. While no has yet criticized (though some have expressed appreciation and/or agreement), I feel that the joke was inappropriate to the venue. My apologies.
Go for it! The "magic" loop is great and makes life sooo much easier. I use it for collecting both the semi-thin sections (instead of an eye lash) and the thins. Not only you get better control of the position of sections on a grid, it also saves you time. Although the loop requires very careful handling - it withstands only certain number of accidents when you bend it from its original angle - it's worth every penny. Good luck, Alice.
-----Original Message----- } From: Claudia Hayward-Costa To: Microscopy-at-sparc5.microscopy.com Sent: 4/25/01 2:57 AM
Dear Microscopists,
I wondered how commonly people use a tool (like a loop) to pick up ultrathin sections.
Does it make life easier or does it only introduce different problems?
I was told that I can buy some "luxury" - but am still undecided.
Your views would be very much appreciated.
Regards
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk Fax: 44(0)208 547 7562
If you are lucky, then you can find the FWBell4080 still with a distributor. However, it was discontiuned. However, you can still get the 4090, which has nice output.
If you also go for the Extech sound meter (as per Nestor's suggestion), then get the high end one. Model 407355 has software for graphing the results. It is well worth the extra $.
JQ
} } From Microscopy-request-at-sparc5.microscopy.com Wed Apr 25 05:22:50 2001 } From: "Anaspec" {anaspec-at-icon.co.za} } To: "'John C. Wheatley'" {John.Wheatley-at-asu.edu} , } {Microscopy-at-sparc5.microscopy.com} } Subject: RE: Search Coil } Date: Wed, 25 Apr 2001 07:46:27 +0200 } } } Hi John } The best bet is to contact your local RS electronics supplier ( find them on } http://www.rs-components.com/ )and ask for the ELF Magnetic Field Strength } meter part number 212 837. } We use it extensively for site tests and have found it to be just as } accurate as using a professional test kit. The only difference between this } unit and a search coil to a scope is that this device simply tells you if } you have a field problem between 20 to 1200Hz where a more expensive search } coil will tell you exactly what frequency it is that is causing the field. } } Good Luck } Luc Harmsen } Anaspec, South Africa } Technical support on microscopy. } Tel + 27 (0) 11 476 3455 } Fax + 27 (0) 11 476 7290 } anaspec-at-icon.co.za } www.anaspec.co.za } } } -----Original Message----- } } From: John C. Wheatley [mailto:John.Wheatley-at-asu.edu] } } Sent: 25 April 2001 01:10 } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Search Coil } } } } } } I need to find a commercial source for a calibrated search coil to check } } stray fields in microscope rooms. Does anyone have any experience with } } purchasing this item? } } } } John C. Wheatley } } Lab Manager } } Arizona State University } } Center for Solid State Science } } PSA-213 } } BOX 871704 } } Tempe, AZ 85287-1704 } } } } } } Phone: (480) 965-3831 } } FAX: (480) 965-9004 } } John.Wheatley-at-ASU.Edu } } } } } } } }
Until recently, I was very much against using a loop to pick up ultrathin sections. A student in the lab started using it and I kind of reluctantly gave it a try. Now I am a regular user of the loop to pick up sections on coated grids.
First I clean the loop with 100% ethanol, air dry and pick up sections. I place a coated grid on a piece of filter paper and bring the loop with the sections down onto the coated grid and let the filter paper soak the water droplet. The sandwiched grids are allowed to air dry for few minutes and you can then remove the loop using a fine tweezer. It works just great.
I bought my loop from Electron Microscopy Sciences. No financial interest with EMS.
Just give it a try.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
On Wed, 25 Apr 2001, Claudia Hayward-Costa wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopists, } } I wondered how commonly people use a tool (like a loop) to pick up } ultrathin sections. } } Does it make life easier or does it only introduce different problems? } } I was told that I can buy some "luxury" - but am still undecided. } } Your views would be very much appreciated. } } Regards } } Claudia } } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } 44(0)208 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk } Fax: 44(0)208 547 7562 } }
In a message dated 4/24/2001 9:41:00 PM Mountain Daylight Time, John.Wheatley-at-asu.edu writes:
} I need to find a commercial source for a calibrated search coil to check } stray fields in microscope rooms. Does anyone have any experience with } purchasing this item?
An inexpensive (~$90) digital gauss meter (Extech Model #480823) that is quite adequate for locating sources of 30 to 300 Hz magnetic fields can be found at Meters and Instruments, "www.MetersandInstruments.com", (800) 773-0370.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Picking up from above is easy with the grid held by a forceps (I prefer a curved Dumont 5 or 7 with a rubber "clamp" to hold it together), the sections just "jump" on my filmed grid. I have tried a Perfect Loop (commercially available) and I also liked it - I could see no difference in "section quality" comparing both methods. Picking up with a grid from below the water surface I do not like (especially with filmed grids, but thats just my personal opinion)
I sometimes loose sections when I am not carefull enough with my eyelash but very very rarely when picking up...
Of course for cryosectioning a loop is an absolute must - and here I clearly prefer the Perfect Loop! The droplet of sucrose solution it can hold is much larger than in a homemade wire loop (gives you a bit more time in the cryochamber) and you can nicely use this large drop as a magnification lens for locating your cryoscetions before picking them up.
Best regards,
Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
This reminds me - we used to get Epon or Spurr's off sandwiched glass slides by putting them in -20C freezer for 1/2 hr and then work the sample off. John Shields EM Lab Univ. of GA Athens
On 24 Apr 2001, at 14:47, Douglas Keene wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } } You may want to try dipping just the thermanox portion of } your block into liquid nitrogen. The sudden change of } temperature will likely loosen the thermanox away from your } sample. I do not expect epon to be a problem, but I do } know that it works well with Spurrs. } } Good luck, } } Doug } } On Mon, 23 Apr 2001 10:12:10 -0500 } "tbargar-at-unmc.edu"-at-sparc5.microscopy.com wrote: } } } } } -------------------------------------------------------------------- } } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } ---. } } } } } } Hi, } } I need advice on embedding Thermanox coverslips. It's } } supposed to peel off leaving the monolayer behind, but I'm } } not having much luck. I'm using Aralidite 502 as the } } embedding medium. Would a another resin work better? I } } would appreciate any and all advice. Thanks. } } } } Tom Bargar } } EM Lab } } UNMC } } 402-559-7347 } } tbargar-at-unmc.edu } } } } } } ---------------------- } Douglas R. Keene } Associate Investigator } Shriners Hospital Research Facilities } 3101 S.W. Sam Jackson Park Road } Portland, Oregon 97201 } phone: 503-221-3434 } FAX: 503-412-6894 (9-5 PST) } e-mail: DRK-at-shcc.org } }
Hi Listers, What's the best 'glue' to use for semiconductor cross section sample prep? We use Gatan G1 to stick the 'sandwich' together (bake at about 150C for an hour) BUT then we stick a 3mm supporting washer onto the mechanically thinned sample using Devcon 5 Minute epoxy prior to ion beam milling. In our constant search for less 'contamination', we are not sure if this is good practice in a FEG (especially) TEM. Are there any other alternative 'glues' that would do the job - without baking? Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
A quick question for the clinical diagnostic people out there. I was wondering what an average turn around time would be for an EM specimen. that is from time of arrival to the time it is placed in the scope.
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} A quick question for the clinical diagnostic people out there. I was {BR} wondering what an average turn around time would be for an EM specimen. that {BR} is from time of arrival to the time it is placed in the scope. {/FONT} {/HTML}
I am trying to find a place to donate my father's microscope. He was a physician, born in Germany. The microscope was bought in Germany, probably in the 1920's or 1930's. It is stored in a wooden case. Is there any place this instrument could be useful? A school or university in the third world? A school in the US?
Please contact me, offline, at ulmithaca-at-home.com.
The Microscopy and Flow Cytometry Core Facility within the MRC Clinical Sciences Centre, Imperial College School of Medicine, has immediate openings for:
Microscopy Research Associate/Assistant (Ref: MRA) Microscopy Core Facility
The imaging of molecules within cells, organs and whole organisms is at the forefront of scientific developments in the life sciences. The Clinical Sciences Centre (CSC) is a leading research institute with groups developing new avenues in the areas of confocal laser scanning microscopy (CLSM), live-cell imaging, scanning ion conductance microscopy, position emission tomography (PET), and nuclear magnetic resonance (NMR). The MRC is seeking a highly motivated individual to be responsible for this state-of-the-art microscope core facility at the CSC, to provide full support for confocal, live-cell imaging, and other fluorescence microscope users within the institute. The successful candidate should have a science degree in a relevant subject. Experience in confocal, live-cell and digital imaging (particularly Leica confocal microscope and/or deconvolution software) would be highly desirable but not essential, as training can be given. Duties will include direct technical support, training and supervision of users. Involvement in research project using fluorescence microscopy will be encouraged (further information about research at the CSC can be found at www.csc.mrc.ac.uk). Informal enquiries and further information about the position to Drs Ana Pombo (tel: +0/44 20 8383 8232; ana.pombo-at-csc.mrc.ac.uk) or Alex Sardini (tel: +0/44 20 8383 8270, a.sardini-at-csc.mrc.ac.uk), MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Campus, Du Cane Road, London W12 0NN. The closing date for applications is 30 April 2001.
Research Assistant (Ref: FCRA) Flow Cytometry Core Facility
Flow cytometric analysis and cell sorting are important for many areas of research in the CSC, including immunology, gene expression and stem cell work. The facility is operated jointly with Imperial College School of Medicine (ICSM) and consists of 2 FACS Vantage cell sorters and several bench top analysers. The MRC is seeking a full time research assistant to help our existing staff provide a cell sorting and analysis service , advise users on the design of their experiments and liase with service engineers (all instruments are on full service contracts). Involvement in research projects will be encouraged. The successful candidate will have a strong interest in the interface between biology and technology. Experience in flow cytometry will be an advantage but is not essential as training can be given. Informal enquiries and further information about the position to Dr Matthias Merkenschlager (tel: +0/44 20 8383 8236/9; matthias.merkenschlager-at-csc.mrc.ac.uk), MRC Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Campus, Du Cane Road, London W12 0NN. The closing date for applications is 30 April 2001.
The CSC is an institute funded by the Medical Research Council and forms part of the ICMS, based at the Hammersmith Hospital in West London. This recently established centre has first class facilities and provides investigators from clinical and basic science backgrounds with the opportunity to pursue innovative, multidisciplinary research within the established clinical base of ICMS. Salaries will be on the MRC's own pay scales and will be commensurate with experience. Further details on how to apply are available from the Human Resources Group tel: 020 8383 3446/7, quoting the relevant reference above.
Advertised in Nature and New Scientist.
Ana Pombo, D.Phil. Nuclear Organisation Group MRC Clinical Sciences Centre Imperial College School of Medicine Hammersmith Hospital Campus Du Cane Road London W12 0NN UK
Although the 1.2MeV HVEM at ANL has sadly departed, we at NCEM want to remind everyone on the listserver that there still exists a HVEM in the US available for on-site and remote use, free of charge with approved proposal.
The Kratos 1.5 MeV HVEM has recently been refurbished (following some substantial high voltage and vacuum issues). It is presently operational at 1 MeV, and is being conditioned for use at 1.5 MeV. 1.5 MeV operation is expected by the end of June, at the latest.
NCEM is actively seeking user proposals for this instrument. Please see:
http://ncem.lbl.gov
http://ncem.lbl.gov/frames/hvem.htm
for further details regarding the instrument's many capabilities (among them straining stage experimentation and environmental cell work), or feel free to contact either myself or Doug Owen - DKOwne-at-LBL.gov
Regards, Eric Stach -- Eric A. Stach Staff Scientist National Center for Electron Microscopy Mail Stop 72-150 Lawrence Berkeley National Laboratory Berkeley, CA 94720 Phone: 510.486.4634 Fax: 510.486.5888 http://ncem.lbl.gov
I am trying to find a place to donate my father's microscope. He was a physician, born in Germany. The microscope was bought in Germany, probably in the 1920's or 1930's. It is stored in a wooden case. Is there any place this instrument could be useful? A school or university in the third world? A school in the US?
Please contact me, offline, at ulmithaca-at-home.com.
Dear Alan, In the original semi-conductor cross-section discussions, M-Bond 610 was recommended. It is a srain-gauge glue, two-part, that requires about 100 deg. C to harden, but it is low viscosity and left a very thin joint. When we were waitng for our order to arrive, we used Devcon 2-ton epoxy, which is higher viscosity and requires eight hours to harden, but still made a thin joint if you squeezed it in a parallel-jawed vice. Once these are hard, we saw no evidence of out-gassing, but we do not have a FEGTEM. At 04:55 PM 4/25/01 +0100, you wrote: } } Hi Listers, } What's the best 'glue' to use for semiconductor cross section sample prep? } We use Gatan G1 to stick the 'sandwich' together (bake at about 150C for an } hour) BUT then we stick a 3mm supporting washer onto the mechanically } thinned sample using Devcon 5 Minute epoxy prior to ion beam milling. } In our constant search for less 'contamination', we are not sure if this is good } practice in a FEG (especially) TEM. Are there any other alternative 'glues' } that would do the job - without baking? } Alan Walker
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Technically you can not remove this type of plastic from a section. Several methods have been published and suggested however, they often destroy the section at the same time. Pam Marcum
-----Original Message----- } From: Rinaldo Pires dos Santos [mailto:rinaldo-at-ufrgs.br] Sent: Wednesday, April 25, 2001 8:23 AM To: Listserv Microscopy
Hi all,
Is there some procedure for to remove hydroxyethylmetacrylate resins (Leica Historesin) from semithin sections (1-2 micrometers)? Thank you.
Dr. Rinaldo Pires dos Santos Lab. of Plant Anatomy - Dept. of Botany E-mail: rinaldop-at-uol.com.br UFRGS - Porto Alegre - RS Brazil
If you are not doing site-specific work with semiconductors as I assume that you are from your message, I would suggest that you use the small angle cleavage technique to do your cross sections. I have done III-V and Si based samples with superb results. Take a look at John McCaffrey and my article in the Number IV MRS TEM Sample Prep book, Vol 480, 1997 for a detailed procedure. South Bay Technology sells the "Micro Cleave Kit". This technique is inexpensive and gives great samples. I have used the samples with no plasma cleaning in a FEGTEM without any problems. In addition, you have no ion milling damage. In fact for a FEG, you have a disadvantage because there is little or no amorphous area to use for focusing.
I use the H-22 silver epoxy from Epoxy Technology to put the samples on to the grids. This requires heating. When I do not want to heat the samples, I have used a Duro 90 minute epoxy for long working times. Unfortunately, it takes about 12 hours to fully cure. I have also used these samples in a FEGTEM without problems. I have cured them in atmosphere and in a nitrogen atmosphere.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: A.Walker [mailto:Alan.Walker-at-sheffield.ac.uk] Sent: Wednesday, April 25, 2001 11:56 AM To: Microscopy-at-sparc5.microscopy.com
Hi Listers, What's the best 'glue' to use for semiconductor cross section sample prep? We use Gatan G1 to stick the 'sandwich' together (bake at about 150C for an hour) BUT then we stick a 3mm supporting washer onto the mechanically thinned sample using Devcon 5 Minute epoxy prior to ion beam milling. In our constant search for less 'contamination', we are not sure if this is good practice in a FEG (especially) TEM. Are there any other alternative 'glues' that would do the job - without baking? Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
The loop works great!!! It's kind of pricy, so careful handling is a must, but it can be a real headache saver. I, too, bought my loop from Electron Microscopy Sciences. I have no financial interest in EMS.
Good luck,
Elizabeth P. Bray Plant Chemist, Central Laboratory South Carolina Electric and Gas Co. 2102 N. Lake Dr. Columbia, SC 29212
----- Original Message ----- } From: "Claudia Hayward-Costa" {LS_S562-at-crystal.kingston.ac.uk} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 25, 2001 5:57 AM
I rather enjoyed it.
Earl
----- Original Message ----- } From: "Chuck Butterick" {cbutte-at-ameripol.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 25, 2001 5:36 AM
SUMMER I 2001 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section B)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 2001 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 29 and end on June 28, 2001.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $92 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum01/away.pdf. The phone registration option is available until tomorrow 4/26/01 (6:30 PM) by calling 516-572-7131 or 7372 or 7425.
P.S. A Fall 2001 TEM course is also being offered (BIO 221 - Section E2) on Thursday evenings beginning at 5:30 PM.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Dr. Rinaldo Pires dos Santos wrote: ================================================================= Is there some procedure for to remove hydroxyethylmetacrylate resins (Leica Historesin) from semithin sections (1-2 micrometers)? Thank you. ================================================================== Perhaps there are other methods, but one that has been used quite successfully has been the use of plasma etching in a unit such as the SPI Plasma Prep™ II.
Some examples are on the SPI Supplies website. For example, on URL http://www.2spi.com/catalog/instruments/etchers1.html you will see an example of a thick section: "Statocyst organ etched 3 min. using O2", and of a thin section, "Low temperature oxygen plasma etched thin section of bacterium embedded in SPI Chem Low Acid GMA for TEM."
Like with any technique in LM or EM, this approach has its limitations. But it is a good way, at least in some instances to remove GMA without disturbing the rest of the sample.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Thanks to all that responded to my question about making it easier for students to cut thin sections using glass knives. Three key points emerged: small block faces, fresh knives (changed often), and a variety of different embedding media. A summary of the responses is below.
Thank you again.
Dick Briggs 1. This is I think a quite common problem. The students in my lab normally embed in Spurr's firm (both animal and plant tissue) and they initially use glass knife and after they feel comfortable with glass knife, I let them use a diamond knife to do their final sectioning. We also use MT2 and MT2B ultramicrotomes.
2. I think LR White resin should fit the bill. It penetrates easily and you don't have the same level of toxicity as with Spurr's (which is always good for beginners).
3. Teaching microtomy is the most difficult portion of any microscopy class. There is such a long learning curve and so many pitfalls that getting good sections depends on the ultimate patience and dexterity of the student. Some will get it right off, others must persevere. You probably already know my suggestions but I'll put my 2 cents in. My suggestion is to start the student with a quality block of a material that cuts easily. Mouse or rat, kidney or liver are good candidates. As they get good sections on these then move them to tissues that may be more difficult. They may be able to troubleshoot problems without questioning the block. I would also ask how the knives are broken? It may be a dull knife problem. A balanced break really helped improve the quality of the knives we were getting from an LKB knife breaker. Additionally a slow break has always seemed better. Wetting the scribe before breaking also lessens the breaking force required and improves edges. Of course small block faces are also going to improve the results dramatically but trying to get the student to cut down the block face is difficult. There are all the usual excuses.
4. I think some of the problem is your choice of tissue rather than plastic. Plants have tough walls, and open spaces mixed together. This causes a lot of different plasticities (?) within a section because some areas will infiltrate to a better or worse degree than a neighboring area. You would have much better luck using a homogeneous mammalian tissue like liver.
5. I faced the same problem. I placed most of the blame on the novices' inability to make good knives. My solution: I break about 25-50 knives a week and give them to the students. I have found that it greatly enhances their ability to get good sections. I also got a grant to buy a new ultramicrotome (Ultracut T). Between my knives and the new 'tome, things are much better now.
6. I think that glass knives dull very rapidly (either damage or plastic build-up on the edge), and so we have always tried to get the first few sections at a given place on the knife edge, which tend to be the best. I wrote a brief procedure for students years ago, based on that approach, and have typed it below (the handout had a drawing of a glass knife, with area A = 1/3 of edge on the side where the whorl meets the edge, area B = middle 1/3 of knife edge, area C = 1/3 of edge at side where whorl is farthest from the edge):
a. Face the block in area C of knife (see diagram). Cut semithin sections (if desired) in area B of knife. Then move to area A. b. Bring the block face parallel with the knife, using the shadow method. Then use the shadow to bring the block face as close to the knife (but without touching) as you dare. c. With the ultramicrotome set for ultrathin sections, manually turn the microtome wheel quite rapidly until you see the first sign of contact (usually a sliver off one side of the block face), then stop turning. d. Turn on automatic sectioning at usual slow cutting speed. Cut about6-12 full-face sections, then stop and pick them up on EM grids. e. Retract the stage slightly, move laterally to another place in area A, and repeat steps 2-4. Continue until area A has all been utilized (or until you have all the sections you need).
7. I teach EM at a four-year college also. I use Spurr's and glass knives made with an LBK 7800 knife maker. I am careful not to select difficult tissues (nervous or muscle or bone or cartilage), but generally my students have been able to get excellent sections by the end of the semester. a. Details to consider: Do your knife maker and microtome work properly (I have an MT2-B and a newer Leica UltraCut. Some students actually prefer the MT2-B. Are you fully dehydrating specimens and are you using truly dry acetone to dehydrate--I dehydrate mine with ashed CUS04. b. Are you buying good float glass and are the students cleaning it fully and handle it carefully. Mine wash with detergent and hot water, DI water, and ethanol, and wash glass and make knives immediately before use.
8. I always have good results using Spurr's. The sample morphology is not as good as other resins. Be sure to polymerize for a good 48 hours or more.
9. I'm doing the same thing with my EM class this semester. I use only Spurr's for both animal and plant tissues and glass knives (our knife breaker is as old as our MT-2). I find students have greater success with the smallest block faces. I also have them use Formvar-coated grids (100 mesh) that tend to stabilize those sections with bad knife marks. I start by making them get thick sections on larger block faces -- they learn the mechanics of the microtomes, block trimming, etc. When they are ready to get thin sections it seems easy to them to just let the motor take over.
10. I typically use an Epon-Araldite mixture for all of my biological samples (generally mammalian soft tissue samples) without any sectioning problems over many years. For botanicals you might need a low viscosity resin like Spurr's. I use MT-2B's in teaching of my students and they rarely have problems obtaining good sections (silver) in ribbons. The key I believe is in the block trimming. My students trim their trapezoid faces no larger than 0.5mm on the longest side (many are around 0.25 mm). I also have the students use the flimsy double-edge razor blades vs. the single edge variety - a trick I learned from my mentor. The blades are much sharper (and more difficult to handle - I require my students to have Band-Aids on hand for block trimming ;-) and give much smoother top and side of the pyramid surfaces. The Epon-Araldite Mixture I use is as follows:
Stock Solution (can be frozen) Small Volume Large Volume Araldite 6005 12.5ml 25ml Poly/Bed 812 15.5ml 31ml Dibutyl phthalate 2ml 4ml
Final Working Solution
Stock Solution 4ml 8ml DDSA 10ml 20ml DMP-30 14 drops* 28 drops* (*Drops introduced with a Pasteur Pipette) 11. Centuries ago, I used the following mixture to embed cellular slime molds for thin-sectioning with glass knives: Araldite 502 13.5 ml DDSA 11.5 ml DMP-30 0.4 ml The Araldite made it a little softer and easier to cut. Hope it helps!
12. My recommendation is to use LR White on a tissue like spleen, kidney, intestine or heart muscle. One mouse would give you more than enough tissue to last a lifetime. Plants are notoriously difficult to section (walls, air spaces, etc.). If you wish to avoid killing animals, you might sacrifice one mouse and put away tissues in fixative or buffer and then provide the students with vials containing the tissues. Also, sometimes vivariums may be euthanizing animals or a fish may die, etc. providing some valuable tissues.
13. When I was an undergraduate (eons ago) I used the "American Araldite" recipe:
Araldite 502 27 parts DDSA 23 parts DMP-30 1.5-2.0% (accelerator) I never included dibutylpthalate.
We mixed it in old (but clean) baby food jars by volume. We had a "reference" jar with marks at the appropriate places and just lined up the reference jar with the one we were measuring into. Very crude but we got good results. I used small pieces of tissue (kidney, spleen, sm. intestine) and went straight from 1:1 to pure resin and into the oven. The mixture is very viscous but can be warmed a bit to make handling easier. I don't know if plant material would work. I used glass knives and an MT-1, but not after a lot of coffee or a night on the town.
14. I have played this game for 32 years now and still don't consider myself an expert because things fool me all the time. I believe almost any embedding medium will work, if instructions are followed and components are fresh. I use pre-mix kits from TAAB, add bottle one to bottle two etc. It's for an easy life. The leftovers, which can be most of the bottle, go into the freezer. Fresh Spurr's cuts like a treat, sometimes thawed Spurr's is fine, sometimes it is not. The reasons can be maybe a little humidity gets in if not up to room temperature (maybe) or it begins to age (increase viscosity due to starting to cure). Anther things to remember are the smaller the block, the better it cut? Also, don't dwell on one piece of edge for too long, often I get just a couple of grids and then move along. I used to make a special mix which was magic (really). It was TAAB-Hard. With about 1% extra MNA (used to weigh all the components) and 1% DMP-30 accelerator. Curing was also important - 48 hours at 35 degrees C, 24 at 45 and 24 at 60 degrees. It was very hard and brittle. It was sterically inhibited i.e. not highly cross-linked but with as much hardener (MNA) as ever allowed. It cut all day on one knife and was terrific. But people didn't want to follow the curing schedule - 4 days!! But the results were the best sections I ever used to cut; I don't do so well these days, even with diamonds.
15. I too teach an undergrad EM course (with Sorvall MT-2s). I've discovered my students are very reluctant to change knives once started. Forcing constant changes has improved the sectioning. We use Spurr's: 10 ERL 5 DER 25 NSA 0.5 DMAE These proportions work best. After dehydration through 50/75/80/95 we go through 4 100% rinses then 1resion:2 ETOH for 3 hours, then 1ETOH to 2 resins (2-3 hours) then full resin overnight - then bake. I assume you're using these proportions so if you want more detail, let me know. The strange thing is my kids do quite well at getting thins with glass knives - they consistently surpass my expectations.
16. We us Epon with mouse liver. Spurr's would probably be better with plants. Plants are always are a tough go for the uninitiated (though probably easier to obtain and no animal right's issues). Plants also require longer infiltration times and smaller increases between infiltration steps (I usually go 25, 50, 70, 80, 90, and a couple of 100% before polymerization). LR White has been suggested, but as the resin is more hydrophilic it is actually more difficult to learn on at first. The block face has a tendency to wet easily and usually students get frustrated trying to figure out why the block face is getting wet even with the more hydrophobic resins like Spurr's or Epon.
17. I find the same problem. I also find that having them get very close to really mastering thick sectioning first (thereby seeing some really pretty LM slides) greatly helps their confidence, and I also have them use the tiniest faces they can manage (0.1mm?) to get started with thins. They must be the correct shape with no ragged edges (such as you might get after taking (or trying to take) many thicks. Also make sure their knives are good - especially FRESH - and that they are using the correct edge and don't' try to take too many sections before switching knives. Have them start with something like liver where the tissue/resin interface is not an issue. I have tried Spurr's but it is so nasty to use, and is not as stable or as pretty in the EM.
Secondarily, I keep troubleshooting sheets taped to the wall - check size! Check shape! Check knife-edge! Check speed! Check angle! etc. I find this helps beginners understand that lots of factors need to be satisfied - it seems that so many of them probably get by in other courses by cramming the night before, which just doesn't' work here... although some of the 'rules' (such as face size) can be relaxed once they get the basics down.
18. There is a resin additive sold by Electron Microscopy Sciences, called "Sure-Cut Surfactant" (their #21630). I have not tried it myself, but remember a posting on this server 1-2 years ago praising this additive very highly. The guy was saying it saved his students a lot of frustration. There is a trade-off, of course, -- they say this additive somewhat interferes with staining. I would give EMS a call; they are usually helpful. (I have no interest in their sales, of course.)
19. Is it the embeddment or sectioning which is the problem? For sectioning try to keep the section area as small as possible. If you keep the sectioning width less than ~.25mm it makes sectioning much easier.
20. There is no magic embedding media that allows easy sectioning with glass knives. All they can do is trim the blocks as small as possible, and I do mean small. If the block face looks big under the high-end magnification of the stereomicroscope it is still too large. Have them practice (a lot) trimming blocks. Don't them rush into thick sections too soon. Make sure they can master the cutting one-micron sections and them trimming to the desired area. The guy I trained under had no clue. He had us cutting as soon as possible. The drop rate for the class was around 50%. And console them even the big folks had to learn using glass knives. It is possible to do. If they think that is hard wait till they stain the sections with Reynolds lead for the first few hundred times. Tell them to cheer up; at least they are not learning on an MT1 manual microtome like I did.
21. Is your course specifically in electron microscopy or would slightly thicker sectioning be almost as good from the microtechnical perspective? For light microscopy, I embed plant tissue in Glycol Methacrylate (GMA) from Electron Microscopy Sciences. The embedding protocol is simpler, and there is the potential that the plastic is less carcinogenic than Spurr's resin. I have recently completed a project in which I was sectioning largish block faces of plant apices embedded in GMA using a glass knife on an Olympus rotary microtome. Sections of less than 1 micrometer were no sweat and I bet they'd be even easier on an ultramicrotome. Very satisfying from the teaching side as it should keep the student frustration level down. GMA is also quite easy to stain with e.g. Richardson's stain or methylene blue. Unfortunately, GMA is a bit too soft to section thinner and it doesn't hold up well under the e-beam so for electron microscopy I use Spurr's or LR White.
22. I noticed your comments on the Microscopy ListServer about students and thin sections. There are a couple of fairly easy ways to make it less painful. My old graduate advisor and I have worked together for many years and I have watched, and at times helped him teach his EM course. For his students sectioning was always the worst experience. Two things might help: 1) use a soft to very soft formulation of one of the Epon substitutes. In that way trimming and thick sectioning are easier and glass knives do not dull as quickly nor do they need to be perfect. The second is make a small block face. Unfortunately that may be more difficult to do than sectioning. Some friends of mine at UC Davis use the soft formulation of Epon which really makes sectioning and trimming easier. It is soft enough to easily make an imprint with a fingernail.
23. I was a former student at San Francisco State University (I've graduated in May 2000), and I took an electron microscopy course at that university. Back to my electron microscopy course. During part of that course, I was supposed to section some embedded tissue of my choice (heart / liver /? (can't recall off-hand)), and I had considerable trouble using a microtome well. As I recall, we had some Sorvalls available for use, but I'm unsure if they were MT-2s. I prepared all my tissues (and practice, plastic "pellets") by first cutting with straight razor blades to make an acceptable-sized pyramid, then continue by sectioning with a microtome. Glass knives were used on these embedded tissues -- the plastic used was Epon -- and I recall that one major problem I had was in making sure that the microtome sectioned at a consistent speed. In addition, the glass knife edges would dull pretty quickly after a small number of sections were made, and so it became imperative to use several glass knives for any particular embedded material. I eventually was able to section pretty well, but I also picked up sections from below with a grid (of 200 mesh) that I bent rather badly during the collection process. Epon worked OK for sectioning, and while it's true that students can indeed get discouraged, I also know from experience that students have very different aptitudes towards handling microtomes, preparing the pyramids, etc. I hope that my experience with sectioning may help you understand better (from a student's perspective) the various trouble areas that could occur when students are first learning how to section embedded material.
********************* 1st Announcement and call for contributions *****************************
A meeting organised by the Royal Microscopical Society and supported by EMAG, FEI UK Ltd, JEOL (UK) Ltd.,LEO EM Inc., Gatan UK, TVIPS GmbH
DEVELOPMENTS IN ENERGY-FILTERED ELECTRON MICROSCOPY
Wednesday 4 July 2001
Department of Materials, Oxford University
The use of electron energy filters in analytical electron microscopy (EFTEM) is a relatively recent development that is proving to be an extremely powerful tool. In-column filters (e.g. so-called "omega-filters") and post-column filters (e.g. the "GIF") represent different approaches which have their own advantages as well as experimental difficulties. This one day meeting is designed to explore the current state of the art, both with regard to the instrumentation and also applications of EFTEM in life and physical sciences.
Invited speakers: Dr Bernd Feja (Tietz Video & Image Processing Systems GbmH) Energy-filtered electron tomography Prof Joachim Mayer (Aachen University of Technology) EFTEM - the state of the art and future trends Dr Paul Midgley (University of Cambridge) EFTEM image series - taking elemental mapping into a new dimension Prof Michael Trendelenburg (German Cancer Research Center, DKFZ) EFTEM in biomedicine & biotechnology: Recent advances in specific element mapping
* CONTRIBUTIONS Contributed presentations are now being sought. Abstracts of no more than 200 words should be sent by email to: crispin.hetherington-at-materials.ox.ac.uk and should arrive by 31 May, 2001.
* REGISTRATION Registration will be £30 RMS/EMAG members, £15 students and £40 others and a printable form will be avialable from the new RMS website, from May on this page :
http://www.rms.org.uk/current%20events.html#eftem
Further details are available from the organisers: Dr Crispin Hetherington, tel. 01865 273799, crispin.hetherington-at-materials.ox.ac.uk Dr Laurence Tetley, tel. 0141 330 4431, l.tetley-at-bio.gla.ac.uk and from the meeting website : http://www-em.materials.ox.ac.uk/events/eftem.html
Note this meeting is timed to follow the 1-day FEGTEM III meeting to be held on 3 July 2001, also in Oxford (http://www-em.materials.ox.ac.uk/events/fegtem.html) **************************************************************************** ****************
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
OK this is going to seem like a stupid question. I am going to try araldite 502 embedding media. the instruction sheet calls for a volumetric measurement. I would prefer to measure the each chemical in a balance. OK here is the question: do i multiply the specific gravity by the volume or divide it. it has been a long time for me since chemistry 101. thanks
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} OK this is going to seem like a stupid question. I am going to try araldite {BR} 502 embedding media. the instruction sheet calls for a volumetric {BR} measurement. I would prefer to measure the each chemical in a balance. OK {BR} here is the question: do i multiply the specific gravity by the volume or {BR} divide it. it has been a long time for me since chemistry 101. {BR} thanks {/FONT} {/HTML}
If I'm not swamped, which is never, I do a two day processing. Using a Lynx overnight to get into resin I then put the specimens into molds and let them cure for 24 hours. When I come in on day three I can cut thick sections, have a conference with the Doc who submitted the case then cut and stain the thins and I'm ready to sit down at the scope. In reality, A specimen coming into our cutting room is submitted for routine light microscopy and possibility special stains. Those sections are then reviewed and the decision is made to submit the case for EM examination. That can take one or two days. So there are four, possibility five days before I can photograph the case. Another issue is prioritization. I do live patients first and I do the tumors before genetics cases. It makes no sense to work on a possible mitochondria disorder when there is a kid with a chest wall tumor or a brain tumor. The same goes for a kidney biopsy over a ciliary disorder. I have to prioritize and frequently cases get put off a few days until the heats down. With a current volume of about four hundred cases a year and an additional two hundred cases for research I'm often asked "How long will this take?" My usual answer which includes the scoping and printing or making a CD is five working days. Autopsy's are not high on my list and periodically I'll hear that the resident who submitted the case has rotated into another department before the EM is done. To answer your question I'd have to know what your case load is and how many hands are available.
Howard Mulhern Supv. Path. EM Facility Children's Hospital Dept. of Pathology 300 Longwood Ave. Boston, 02115 Ma.
********************* 1st Announcement and call for contributions *****************************
A meeting organised by the Royal Microscopical Society and supported by EMAG, FEI UK Ltd, JEOL (UK) Ltd.,LEO EM Inc., Gatan UK, TVIPS GmbH
DEVELOPMENTS IN ENERGY-FILTERED ELECTRON MICROSCOPY
Wednesday 4 July 2001
Department of Materials, Oxford University
The use of electron energy filters in analytical electron microscopy (EFTEM) is a relatively recent development that is proving to be an extremely powerful tool. In-column filters (e.g. so-called "omega-filters") and post-column filters (e.g. the "GIF") represent different approaches which have their own advantages as well as experimental difficulties. This one day meeting is designed to explore the current state of the art, both with regard to the instrumentation and also applications of EFTEM in life and physical sciences.
Invited speakers: Dr Bernd Feja (Tietz Video & Image Processing Systems GbmH) Energy-filtered electron tomography Prof Joachim Mayer (Aachen University of Technology) EFTEM - the state of the art and future trends Dr Paul Midgley (University of Cambridge) EFTEM image series - taking elemental mapping into a new dimension Prof Michael Trendelenburg (German Cancer Research Center, DKFZ) EFTEM in biomedicine & biotechnology: Recent advances in specific element mapping
* CONTRIBUTIONS Contributed presentations are now being sought. Abstracts of no more than 200 words should be sent by email to: crispin.hetherington-at-materials.ox.ac.uk and should arrive by 31 May, 2001.
* REGISTRATION Registration will be £30 RMS/EMAG members, £15 students and £40 others and a printable form will be avialable from the new RMS website, from May on this page :
http://www.rms.org.uk/current%20events.html#eftem
Further details are available from the organisers: Dr Crispin Hetherington, tel. 01865 273799, crispin.hetherington-at-materials.ox.ac.uk Dr Laurence Tetley, tel. 0141 330 4431, l.tetley-at-bio.gla.ac.uk and from the meeting website : http://www-em.materials.ox.ac.uk/events/eftem.html
Note this meeting is timed to follow the 1-day FEGTEM III meeting to be held on 3 July 2001, also in Oxford (http://www-em.materials.ox.ac.uk/events/fegtem.html) **************************************************************************** ****************
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
********************* 1st Announcement and call for contributions *****************************
A meeting organised by the Royal Microscopical Society and supported by EMAG, FEI UK Ltd, JEOL (UK) Ltd.,LEO EM Inc., Gatan UK, TVIPS GmbH
DEVELOPMENTS IN ENERGY-FILTERED ELECTRON MICROSCOPY
Wednesday 4 July 2001
Department of Materials, Oxford University
The use of electron energy filters in analytical electron microscopy (EFTEM) is a relatively recent development that is proving to be an extremely powerful tool. In-column filters (e.g. so-called "omega-filters") and post-column filters (e.g. the "GIF") represent different approaches which have their own advantages as well as experimental difficulties. This one day meeting is designed to explore the current state of the art, both with regard to the instrumentation and also applications of EFTEM in life and physical sciences.
Invited speakers: Dr Bernd Feja (Tietz Video & Image Processing Systems GbmH) Energy-filtered electron tomography Prof Joachim Mayer (Aachen University of Technology) EFTEM - the state of the art and future trends Dr Paul Midgley (University of Cambridge) EFTEM image series - taking elemental mapping into a new dimension Prof Michael Trendelenburg (German Cancer Research Center, DKFZ) EFTEM in biomedicine & biotechnology: Recent advances in specific element mapping
* CONTRIBUTIONS Contributed presentations are now being sought. Abstracts of no more than 200 words should be sent by email to: crispin.hetherington-at-materials.ox.ac.uk and should arrive by 31 May, 2001.
* REGISTRATION Registration will be £30 RMS/EMAG members, £15 students and £40 others and a printable form will be avialable from the new RMS website, from May on this page :
http://www.rms.org.uk/current%20events.html#eftem
Further details are available from the organisers: Dr Crispin Hetherington, tel. 01865 273799, crispin.hetherington-at-materials.ox.ac.uk Dr Laurence Tetley, tel. 0141 330 4431, l.tetley-at-bio.gla.ac.uk and from the meeting website : http://www-em.materials.ox.ac.uk/events/eftem.html
Note this meeting is timed to follow the 1-day FEGTEM III meeting to be held on 3 July 2001, also in Oxford (http://www-em.materials.ox.ac.uk/events/fegtem.html) **************************************************************************** ****************
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
One thing that was not mentioned was the use of resin conditioners that help to preserve the knife edge of a glass knife. We use this with beginners. We have used lecithin and "PolyCut Ease" from Polysciences Information on this from a previous discussion can be found at http://www.biotech.ufl.edu/icbr/emcl/db/slippery.html I am not sure if a conditioner for LR White is mentioned there or in the Mollenhauer pub., but in my personal conversations with Hilton, he said that camphor could be used for acrylic resins. If this is of any interest I can dig deeper into this or perhaps we can contact Hilton Mollenhauer directly.
At 07:25 AM 4/26/2001 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greg Erdos Assistant Director Biotechnology Program Ph. 352-392-1295 University of Florida Fax 352-846-0251 PO Box 118525 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
} If I'm not swamped, which is never, I do a two day processing. Using } a Lynx overnight to get into resin. I then put the specimens into molds and } let them cure for 24 hours. When I come in on day three I can cut thick } sections, have a conference with the Doc who submitted the case then cut } and stain the thins and then I'm ready to sit down at the scope.
} In reality, A specimen coming into our cutting room is submitted for } routine light microscopy and possibility special stains. Those sections are } then reviewed and the decision is made to submit the case for EM } examination. That can take one or two days. So there are four, possibility } five days before I can photograph the case. Another issue is } prioritization. I do live patients first and I do the tumors before } genetics cases. It makes no sense to work on a possible mitochondria } disorder when there is a kid with a chest wall tumor or a brain tumor. The } same goes for a kidney biopsy over a ciliary disorder. I have to prioritize } and frequently cases get put off a few days until the heats down. With a } current volume of about four hundred cases a year and an additional two } hundred cases for research I'm often asked "How long will this take?" My } usual answer which includes the scoping and printing or making a CD is five } working days. Autopsy's are not high on my list and periodically I'll hear } that the resident who submitted the case has rotated into another } department before the EM is done. To answer your question I'd have to know } what your case load is and how many skilled hands are available.
} Howard Mulhern } Supv. Path. EM Facility } Children's Hospital } Dept. of Pathology } 300 Longwood Ave. } Boston, 02115 Ma. } } } }
I have been using a Chien grid to pick up sections, like a loop, and place it on top of a coated grid. Let water evaporate and sections will drop on to the grid below. I dip Chien grids in 1% formvar, shake off excess and place then on a filter paper to dry. This treatment ensures water film is maintained during the transfer. Cleaning the Chien grids in acid has the same effect. The handle of Chien grid is bended for better handling. I have tried a "perfect loop" and I prefer Chien grids.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Claudia Hayward-Costa {LS_S562-at-crystal.kingston.ac.uk} 04/25 5:57 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Microscopists,
I wondered how commonly people use a tool (like a loop) to pick up ultrathin sections.
Does it make life easier or does it only introduce different problems?
I was told that I can buy some "luxury" - but am still undecided.
Your views would be very much appreciated.
Regards
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk Fax: 44(0)208 547 7562
} -----Original Message----- } From: Earl Weltmer [SMTP:eweltmer-at-home.com] } Sent: Wednesday, April 25, 2001 8:19 PM } To: Chuck Butterick; Microscopy-at-sparc5.microscopy.com } Subject: Re: Apology } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I rather enjoyed it. } } Earl } } ----- Original Message ----- } } From: "Chuck Butterick" {cbutte-at-ameripol.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, April 25, 2001 5:36 AM } Subject: Apology } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Listers, } } } } A slip of the finger caused me to send an un-topical joke to all. } } While no has yet criticized (though some have expressed } appreciation } } and/or agreement), I feel that the joke was inappropriate to the } } venue. My apologies. } } } } Chuck Butterick } } } } }
OK this is going to seem like a stupid question. I am going to try araldite 502 embedding media. the instruction sheet calls for a volumetric measurement. I would prefer to measure the each chemical in a balance. OK here is the question: do i multiply the specific gravity by the volume or divide it. it has been a long time for me since chemistry 101. thanks
Dear J, At the beginning of every semester I give three general hints on calculations to my students. The relevant one for you is always write down units explicitly. Since the density (numerically equal to the specific gravity in cgs units) is in g/cm^3, if you multiply the volume (in cm^3) by the density, you get the mass. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Following the recent discussion thread of scanners, I wanted to say that we are the proud owners of a Polaroid SprintScan 45 Ultra, and are very happy with the results we have gotten from it. It came with a number of negative holders covering a range of sizes from 35mm to 4x5, except one for 3.25x4.25 TEM sheet film. Right now we're trying to think of ways around this problem and would be happy to hear offline from anyone who has some ideas and/or solutions. I would even entertain the idea of having a holder made (this is not an option locally for me).
Thanks in advance for any help (and there is usually plenty, thanks to this list!).
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Tel: (902) 679-5566 FAX: (902) 679-2311
email: allanwojtasp-at-em.agr.ca
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Here in the lab we still do the hand processing.. not automated processor yet.
There are two techs, but turnaround time is different between both of us.. i.e. one of us works faster than the other...
Normally when a biopsy comes in on say Monday.. I can have images by Wednesday.. Out here we do not use a darkroom anymore.. Our lab has gone digital with a AMT system.
The Pathologists are thrilled with it since it cuts down our turnaround time from 5 days to 3 days...
Does anyone knows where I can find a five-axes (four-axes U-Stage is fine too) universal stage for sale or donation?
Thank all of you in advance.
Best wishes,
Leonardo -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
I would like to start up a thread about the level of instruction and the amount of time that undergraduates spend in an introductory EM course that covers TEM & SEM. The need comes from low numbers of student credit hours generated verses the amount of time spent by the students and instructor in the course. For example, I usually have between 2 to 4 students in my four credit hour course, taught every fall sememster. The students receive two one hour lectures per week, two supervised two hour labs and a third unsupervised two hour lab. I teach the students in pairs during the intensive hands-on portions like ultramicrotomy and scope operations, so I usually have two lab sections to prep for, or about eight contact hours for labs per week. With the lectures, I then have ten hours of contact per week for the course.
What are the teaching loads and class sizes of other instructors who teach undergrad EM?
Thanks
Robert
Robert Fitton Teaching Associate/Director of Laboratories Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 563-387-1559 FAX 563-387-1080 (If the 563 area code does not work, try our old area code of 319)
Enjoy a visit to our website: http://www.luther.edu/~biodept/
Our TAT differs depending on the service rendered.
Diagnostic virology: 15 min to ~2 hours (depending on whether we ultracentrifuge) for negative staining of bodily fluids. 2-3 days for thin sections of tissue (depending on how many blocks we have to cut and how long we have to look if the sample shows no viruses before we call it negative). Three super techs do ~700 negative stain specimens and 100 thin section samples of clinical material per year, plus a number of research samples (20-40, some of which are cryosections for IEM).
Surgical pathology: 2 days (about 32 hours). In on a morning, into the oven that night, cut, photographed, and dark room micrographs printed and delivered to the pathologist by 5 PM the next day. (We're working on going digital.) Two super techs do ~500 clinical samples plus ~100 research samples a year.
Five techs total. They can cross cover, but each has a specialty, and generally, the same 3 do virology, and the other 2 do surgical pathology.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Dear Microscopist,
} Does anyone knows where I can find a five-axes (four-axes U-Stage is } fine too) universal stage for sale or donation?
} Thank all of you in advance.
} Best wishes,
} Leonardo } -- } --- } Leonardo Lagoeiro } Departamento de Geologia } Universidade Federal de Ouro Preto } Ouro Preto, MG, 35400-000 } Brazil } E-mai: lagoeiro-at-degeo.ufop.br
The message you sent to jeolbxl-at-pophost.eunet.be could not be delivered. Either you misspelled your correspondent's name, or jeolbxl no longer exists in the pophost.eunet.be domain.
If you want more information, you can contact the postmaster at postmaster-at-pophost.eunet.be. Please understand that we can not give emailaddresses from our customers.
Kind Regards,
The mail delivery system. -- --- Leonardo Lagoeiro Departamento de Geologia Universidade Federal de Ouro Preto Ouro Preto, MG, 35400-000 Brazil E-mai: lagoeiro-at-degeo.ufop.br
Polaroid has a small sheet insert that fits into the 4 X 5 and holds the negative down with small magnets. You need to call someone at Polaroid and request this. They gave them to me and a few others on the List for free. I originally talked to John Warren WARRENJ1-at-POLAROID.COM There is also a Dennis Lizier, but I do not have his contact information. -Scott
-----Original Message----- } From: Paula Allan-Wojtas [mailto:AllanWojtasP-at-em.agr.ca] Sent: Thursday, April 26, 2001 12:17 PM To: microscopy-at-sparc5.microscopy.com
Hi, all,
Following the recent discussion thread of scanners, I wanted to say that we are the proud owners of a Polaroid SprintScan 45 Ultra, and are very happy with the results we have gotten from it. It came with a number of negative holders covering a range of sizes from 35mm to 4x5, except one for 3.25x4.25 TEM sheet film. Right now we're trying to think of ways around this problem and would be happy to hear offline from anyone who has some ideas and/or solutions. I would even entertain the idea of having a holder made (this is not an option locally for me).
Thanks in advance for any help (and there is usually plenty, thanks to this list!).
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Tel: (902) 679-5566 FAX: (902) 679-2311
email: allanwojtasp-at-em.agr.ca
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Robert Any undergraduate teaching of EM techniques we do strictly within the context of student projects, chiefly in the final honours year (4th year of the BSc course at Edinburgh). Since I joined the Botany department staff in 1976 the policy has been not to teach methods. I don't especially agree with that policy, but it is the way things have always been done here. I get the luxury of a 1-hour presentation on EM to first year biology students at the end of their first year. We also demonstrate the EMs to class groups from various departments, and we train a few students per year as they require access to EM for project work, but there have been no formal taught undergraduate courses with hands-on practical tuition in EM in the Science Faculty at Edinburgh for many years now. Chris
----- Original Message ----- } From: "Robert Fitton" {fittonro-at-luther.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 26, 2001 9:40 PM
FW: Re: Film Processing and dynamic range and Exposure of film to electrons
} A couple of thoughts on film: } Kodak special publication P-116 describes the interaction of electrons with film (from the point of view of people that produce the film-I assume they study it fairly closely): } "Each electron in an incident beam may pass through perhaps 30 or more silver halide grains before it is stopped completely...........The trajectory is a changing one as collisions occur, and energy is lost to the silver halide grains, and to a lesser extent, to the gelatin in which the silver halide grains are dispersed. This transfer of energy, which is low at the beginning of an electron path and increases as the electron is slowed by collisions, is responsible for the formation of specs of silver atoms in the affected grain. If the aggregate of silver atoms formed is sufficiently large-believed to be between 3 and 6 atoms, the entire grain will be capable of being converted to metallic silver during photographic development. While the passage of a single electron may not render each grain developable, to which it gives up energy, overall sensitivity is such that normally at least one grain, and very likely, a number of grains, will be converted to metallic silver. If this is the case, the photographic material records all of the information in the electron beam;" } This document further discusses contrast, illustrating the sensitivity of the film with log exposure/density curves, but only up to a density of 2. It may be worth noting that John Spence states in the appendix of his text that for photographic emulsions, the exposure/density curve is linear up to a density of between one and 2. The departure from linearity would seem to be related to some sort of "pile-up" effect. While the exposure/density curves may be linear over this range, the read-out of the film is logarithmic, as the density is defined as the log base 10 of the transmitted light intensity divided by the incident light intensity. That is why, for example, one commonly used, and successful method of increasing contrast is to increase the exposure level of the film. For example, a 10% change in electron dose near a density of 1 will produce a 25% change in transmitted light intensity, where a 10 % change in dose near a density of 2 will produce a 58% change in transmitted light intensity. This logarithmic read-out characteristic is perhaps why all of the Kodak sensitivity plots are in log dose/density format, and they are curves. Kodak also illustrates how the contrast of the print can be estimated from the slope of these log dose/density curves, since the slope increases with average density. In summary, even though the density of the film may be linear with electron dose, what you see in a print, what you get from a scanner, even what you see from observing the film on a light box, is logarithmic with respect to electron dose. This is also why images obtained with a CCD camera or an image plate, which are linear with electron dose, have a distinctly different appearance than images obtained with film unless digital gamma is used to adjust the display. So, everyone was essentially correct in what they said, it's just that they were not describing the same thing. } One may want to look at the claims made for Fuji image plates on their web site, which can be accessed at "fujimed.com/sub/FDL5000.pdf". They show plots of log readout intensity/log dose for the Fuji image plate superimposed on density/log dose for film. The Fuji image plate is more linear than film, but for the range 0-2, film is relatively parallel to the image plate. Beyond a density of 2, the nonlinearity becomes gradually more pronounced, but there is never any saturation encountered. } Finally, I would like to reiterate that this has been a fascinating string, and it seems that everyone has contributed some useful and thought provoking information.} } } John Mardinly } Intel Materials Technology
I have the pleasure of teaching full semester (13 week) EM course to undergraduates at Queensland University of Technology in Brisbane, Australia. We've offered this course for quite a number of years - in fact, even had it extended from part of a broader topic to a full semester just for EM! The course is offered as part of the School of Life Sciences undergraduate program and, hence, primarily addresses biological EM. However, it is open to students from other disciplines - presently we do not have a full EM course in the physical sciences, although my colleague Dr Thor Bostrom teaches several weeks of EM in various courses for physics, chemistry and engineering undergraduate students.
Apart from this, I will usually run single 2 hour practical sessions for a couple of other undergraduate courses (mainly microbiology, where students look at negatively stained microbes they have isolated from various sources). I'll often be asked to give a half hour (sometimes an hour) introductory EM lecture to these students, prior to the practicals.
I also teach a brief introduction (2-3 weeks) to EM as part of a post-graduate course on cell structure and function in pathology. Other post-graduates needing EM as part of their research are taught individually by staff in our EM Facility, to the level that their projects require.
My full semester EM course is offered in the third year of the B. Applied Science course. This year I have 30 students - the numbers have been increasing over the past few years from a previous average of 20. I usually also have a couple of post-graduate (research) students attending the lectures - these students will be using our EM facilities as part of their research, and I find this is an effective means of giving them some useful background and theory to support their laboratory work.
For this course, I am allocated two by one hour lectures each week (total of 26 hours lectures for the semester). I cover basic instrumentation (TEM, SEM, preparation equipment) and biological sample preparation methods, including cryotechniques. I also include immunolabelling methods, some cytochemistry, microanalysis (Thor Bostrom gives these lectures), a few interesting "non-biological" applications, and a brief introduction to some other imaging methods (confocal, AFM etc).
Practical sessions have been a bit of a "challenge" as student numbers have increased and EM staff numbers have decreased! However, I've now got a system that works, and doesn't cause me (or the equipment) too much stress. I have two by 3 hours practical sessions running each week - ie. 6 hours of my teaching time per week (total of 72 hours of my teaching time for the semester).
Each student attends a total of 24 hours practical sessions for the semester (this fits with our "official" allocation for total practical time for a course of this credit point value). I divide the class into small groups - 3 to 4 students per group, with current student numbers. For preparation lab based pracs (where we have enough space and the students don't need such close supervision), I'll have a number of these small groups attending the same practical session. For EM operation and viewing pracs, I'll only have one small group attending at a time (usually 1 or 1.5 hours of the session for each small group). This has enabled me to run practical sessions almost single-handed - money to employ tutors/demonstrators or other teaching assistants has been a bit difficult to obtain! If anyone would like further information on the practicals I run or on the logistics of this, please contact me by direct email - does anyone else out there run EM practicals for undergraduates??? I haven't seen a rush of emails in response to Robert's questions.
Fortunately, my university is still keen to have students doing "hands-on" practicals in their courses. However, I've certainly had to decrease practical work substantially since I first started teaching in this EM course. A lot of this is due to decreased staff numbers in our EM Facility - a problem that I guess most of us face. I teach other courses (microbiology, parasitology) and encounter the same problems there - it is now very difficult to get competent, experienced people to assist in practical classes, particularly for more advanced or specialised courses.
On the bright side, I've found the students enjoy EM a great deal - and it is one of the few opportunities our undergraduates get to use expensive and (relatively!) modern instrumentation, even if it is only on a very limited scale. I've not had many students who don't show a great deal of enthusiasm when told it is now their turn to operate the TEM or SEM and take some micrographs. This semester, more so than previously, at the end of each practical session quite a number of my students have taken the time to thank me for running the practicals (even after the night classes which finish at 8pm). I figure that has got to be a good sign!!!
Regards Deb ***************************************************** Dr Deborah Stenzel Lecturer (Microbiology) School of Life Sciences and Applications Specialist (Biological) Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434 Brisbane 4001 Australia
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Hi, everyone, Now, I need to simulate HRTEM image of a phase that I can not find its space group in X-ray powder diffraction files. Do you know whether it is possible for me to find its atomic positions. Could you please give me suggestions? Thank you very much in advance. WY
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Dear Listers, We are in need to prepare a short course on EM basics. I would be thankfull for any information about free internet resources or commercially availiable CDs containing illustative materials, animations, programms illustrating image formation (not simulating software) that we can use in the lectures and during the seminars.
EuroFE 2001, the European Meeting on Applications of Field Emission Technologies, EuroFE 2001 will tale palace this year in University of Alicante, Spain from November 12-16, 2001. Details are available at www.cmp-cientifica.com/eurofe2001
The fundamental purpose of workshops such as EUROFE2001 consists in bringing together a hundred scientific, leaders in this area, in order to: Link all active research groups in Field Emission.
Link all industries interested in Field Emission. At EUROFE2001, major companies such as Motorola (USA), Candescent (USA), Samsung (Korea), PixTech (France), Saint-Gobain (France), Thomson (France), LG (Korea) and PFE (UK) will be present.
Foster co-operation and the interchange of ideas between research groups and European Industry.
Provide an overview of the research and commercialisation of Field Emission technologies within Europe. During EUROFE2001, a session will be dedicated to Space Applications (ESA/ESTEC speakers).
Allow rapid and flexible response to new technological challenges
Based on the format established at EUROFE2000, ample time for discussion will be available for meaningful interaction between senior researchers/graduate students and industrial partners.
Funds to offset travel expenses for graduate students to present their work in a poster session at EUROFE2001 will be available.
For further details please contact:
Antonio Correia CMP Cientifica Apdo. Correos 20 28230 Las Rozas (Madrid), Spain Tel: +34 91 6407187 Fax: +34 91 6407186
Regards,
Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
Hi, everyone, Now, I need to simulate HRTEM image of a phase that I can not find its space group in X-ray powder diffraction files. Do you know whether it is possible for me to find its atomic positions. Could you please give me suggestions? Thank you very much in advance. WY
Carbon black in polymer or rubber will cause no more problems than most any other hydrocarbon. The electron beam does sublimate the plastic. Especially when working with pure carbon black, one can essentially watch a contaminant layer grow on the constituent particles of a given aggregate. A cold finger is a very good idea but not absolutely required if you are just scanning the sample.
Our Philips 300 is still in excellent operating condition after 28 years of carbon black work.
Chuck Butterick Engineered Carbons, Inc. Borger, TX
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id HAA26160 for dist-Microscopy; Fri, 27 Apr 2001 07:37:15 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id HAA26157 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 27 Apr 2001 07:36:45 -0500 (CDT) Received: from mercury.uwe.ac.uk (mercury.uwe.ac.uk [164.11.132.23]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id HAA26150 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 27 Apr 2001 07:36:33 -0500 (CDT) Received: from fas655.uwe.ac.uk ([164.11.205.145]) by mercury.uwe.ac.uk (2.0.4/SMS 2.0.4-devel) with SMTP id NAA04426; Fri, 27 Apr 2001 13:33:48 +0100 (BST)
Back in 1989 all Biology degree and sub-degree students attended 3 sessions in the EM Unit. Now the expansion in numbers precludes such demonstrations. I regret this change.
In the first year they get a few lectures and a "dry" class identifying cell organelles. In the final year about 6 students do major projects in the lab.
We still do undergraduate classes in dust ID and in EDX for modules taken by Chemistry, Environmental Science and Environmental Health students as the numbers are smaller.
Dave
On Fri, 27 Apr 2001 16:08:08 +1000 Dr Deborah Stenzel {d.stenzel-at-qut.edu.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Robert and all } } I have the pleasure of teaching full semester (13 week) EM course to } undergraduates at Queensland University of Technology in Brisbane, } Australia. We've offered this course for quite a number of years - in } fact, even had it extended from part of a broader topic to a full semester } just for EM! The course is offered as part of the School of Life Sciences } undergraduate program and, hence, primarily addresses biological } EM. However, it is open to students from other disciplines - presently we } do not have a full EM course in the physical sciences, although my } colleague Dr Thor Bostrom teaches several weeks of EM in various courses } for physics, chemistry and engineering undergraduate students. } } Apart from this, I will usually run single 2 hour practical sessions for a } couple of other undergraduate courses (mainly microbiology, where students } look at negatively stained microbes they have isolated from various } sources). I'll often be asked to give a half hour (sometimes an hour) } introductory EM lecture to these students, prior to the practicals. } } I also teach a brief introduction (2-3 weeks) to EM as part of a } post-graduate course on cell structure and function in pathology. Other } post-graduates needing EM as part of their research are taught individually } by staff in our EM Facility, to the level that their projects require. } } } My full semester EM course is offered in the third year of the B. Applied } Science course. This year I have 30 students - the numbers have been } increasing over the past few years from a previous average of 20. I } usually also have a couple of post-graduate (research) students attending } the lectures - these students will be using our EM facilities as part of } their research, and I find this is an effective means of giving them some } useful background and theory to support their laboratory work. } } For this course, I am allocated two by one hour lectures each week (total } of 26 hours lectures for the semester). I cover basic instrumentation } (TEM, SEM, preparation equipment) and biological sample preparation } methods, including cryotechniques. I also include immunolabelling methods, } some cytochemistry, microanalysis (Thor Bostrom gives these lectures), a } few interesting "non-biological" applications, and a brief introduction to } some other imaging methods (confocal, AFM etc). } } Practical sessions have been a bit of a "challenge" as student numbers have } increased and EM staff numbers have decreased! However, I've now got a } system that works, and doesn't cause me (or the equipment) too much } stress. I have two by 3 hours practical sessions running each week - ie. 6 } hours of my teaching time per week (total of 72 hours of my teaching time } for the semester). } } Each student attends a total of 24 hours practical sessions for the } semester (this fits with our "official" allocation for total practical time } for a course of this credit point value). I divide the class into small } groups - 3 to 4 students per group, with current student numbers. For } preparation lab based pracs (where we have enough space and the students } don't need such close supervision), I'll have a number of these small } groups attending the same practical session. For EM operation and viewing } pracs, I'll only have one small group attending at a time (usually 1 or 1.5 } hours of the session for each small group). This has enabled me to run } practical sessions almost single-handed - money to employ } tutors/demonstrators or other teaching assistants has been a bit difficult } to obtain! If anyone would like further information on the practicals I } run or on the logistics of this, please contact me by direct email - does } anyone else out there run EM practicals for undergraduates??? I haven't } seen a rush of emails in response to Robert's questions. } } Fortunately, my university is still keen to have students doing "hands-on" } practicals in their courses. However, I've certainly had to decrease } practical work substantially since I first started teaching in this EM } course. A lot of this is due to decreased staff numbers in our EM Facility } - a problem that I guess most of us face. I teach other courses } (microbiology, parasitology) and encounter the same problems there - it is } now very difficult to get competent, experienced people to assist in } practical classes, particularly for more advanced or specialised courses. } } On the bright side, I've found the students enjoy EM a great deal - and it } is one of the few opportunities our undergraduates get to use expensive and } (relatively!) modern instrumentation, even if it is only on a very limited } scale. I've not had many students who don't show a great deal of } enthusiasm when told it is now their turn to operate the TEM or SEM and } take some micrographs. This semester, more so than previously, at the end } of each practical session quite a number of my students have taken the time } to thank me for running the practicals (even after the night classes which } finish at 8pm). I figure that has got to be a good sign!!! } } } Regards } Deb } ***************************************************** } Dr Deborah Stenzel } Lecturer (Microbiology) } School of Life Sciences } and } Applications Specialist (Biological) } Analytical Electron Microscopy Facility } Queensland University of Technology } GPO Box 2434 } Brisbane 4001 } Australia } } Phone + 61 7 3864 5036 } Fax + 61 7 3864 5100 } email d.stenzel-at-qut.edu.au } } http://www.sci.qut.edu.au/aemf } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
You make mention of "the EM staff" - I'm wondering how many people are actually involved in teaching the course components, and how much time do they collectively spend, for example per student per week??
I'm curious to know because we have two people involved in teaching a full semester Bio EM course for up to 8 students. The students in the end turn in a fairly polished portfolio (with about a dozen 8x10 micrographs plus figure legends, from three different tissues that they have processed, cut, and scoped).
Questions have recently arisen as to how many man-hours we 'should' allot to such a course, which is far outside the norm for our other lab courses.
I'm interested in off-line dialogue, if others are.
Ann Hein Lehman EM Facility Manager Trinity College Hartford CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu
-----Original Message----- } From: Dr Deborah Stenzel [mailto:d.stenzel-at-qut.edu.au] Sent: Friday, April 27, 2001 2:08 AM To: Microscopy-at-sparc5.microscopy.com
Dear Robert and all
I have the pleasure of teaching full semester (13 week) EM course to undergraduates at Queensland University of Technology in Brisbane, Australia. We've offered this course for quite a number of years - in fact, even had it extended from part of a broader topic to a full semester just for EM! The course is offered as part of the School of Life Sciences undergraduate program and, hence, primarily addresses biological EM. However, it is open to students from other disciplines - presently we do not have a full EM course in the physical sciences, although my colleague Dr Thor Bostrom teaches several weeks of EM in various courses for physics, chemistry and engineering undergraduate students.
Apart from this, I will usually run single 2 hour practical sessions for a couple of other undergraduate courses (mainly microbiology, where students look at negatively stained microbes they have isolated from various sources). I'll often be asked to give a half hour (sometimes an hour) introductory EM lecture to these students, prior to the practicals.
I also teach a brief introduction (2-3 weeks) to EM as part of a post-graduate course on cell structure and function in pathology. Other post-graduates needing EM as part of their research are taught individually by staff in our EM Facility, to the level that their projects require.
My full semester EM course is offered in the third year of the B. Applied Science course. This year I have 30 students - the numbers have been increasing over the past few years from a previous average of 20. I usually also have a couple of post-graduate (research) students attending the lectures - these students will be using our EM facilities as part of their research, and I find this is an effective means of giving them some useful background and theory to support their laboratory work.
For this course, I am allocated two by one hour lectures each week (total of 26 hours lectures for the semester). I cover basic instrumentation (TEM, SEM, preparation equipment) and biological sample preparation methods, including cryotechniques. I also include immunolabelling methods, some cytochemistry, microanalysis (Thor Bostrom gives these lectures), a few interesting "non-biological" applications, and a brief introduction to some other imaging methods (confocal, AFM etc).
Practical sessions have been a bit of a "challenge" as student numbers have increased and EM staff numbers have decreased! However, I've now got a system that works, and doesn't cause me (or the equipment) too much stress. I have two by 3 hours practical sessions running each week - ie. 6 hours of my teaching time per week (total of 72 hours of my teaching time for the semester).
Each student attends a total of 24 hours practical sessions for the semester (this fits with our "official" allocation for total practical time for a course of this credit point value). I divide the class into small groups - 3 to 4 students per group, with current student numbers. For preparation lab based pracs (where we have enough space and the students don't need such close supervision), I'll have a number of these small groups attending the same practical session. For EM operation and viewing pracs, I'll only have one small group attending at a time (usually 1 or 1.5 hours of the session for each small group). This has enabled me to run practical sessions almost single-handed - money to employ tutors/demonstrators or other teaching assistants has been a bit difficult to obtain! If anyone would like further information on the practicals I run or on the logistics of this, please contact me by direct email - does anyone else out there run EM practicals for undergraduates??? I haven't seen a rush of emails in response to Robert's questions.
Fortunately, my university is still keen to have students doing "hands-on" practicals in their courses. However, I've certainly had to decrease practical work substantially since I first started teaching in this EM course. A lot of this is due to decreased staff numbers in our EM Facility - a problem that I guess most of us face. I teach other courses (microbiology, parasitology) and encounter the same problems there - it is now very difficult to get competent, experienced people to assist in practical classes, particularly for more advanced or specialised courses.
On the bright side, I've found the students enjoy EM a great deal - and it is one of the few opportunities our undergraduates get to use expensive and (relatively!) modern instrumentation, even if it is only on a very limited scale. I've not had many students who don't show a great deal of enthusiasm when told it is now their turn to operate the TEM or SEM and take some micrographs. This semester, more so than previously, at the end of each practical session quite a number of my students have taken the time to thank me for running the practicals (even after the night classes which finish at 8pm). I figure that has got to be a good sign!!!
Regards Deb ***************************************************** Dr Deborah Stenzel Lecturer (Microbiology) School of Life Sciences and Applications Specialist (Biological) Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434 Brisbane 4001 Australia
Like Chris, our department does not teach techniques courses. However, I include SEM and TEM in a course called Microanatomy (about 16 students every fall semester). What I have done is combine more-or-less traditional histology class lectures with a lab component in which students compare the histology of the same organ from a mouse and frog. They make their own paraffin, methacrylate, and epoxy plastic sections and critical point dry their other samples. I spend a great deal of time on light microscopy theory, how to clean and align a light microscope, capture digital images, and prepare micrographs for illustration (scale bars and labels). I do not spend a lot of time on the use of the electron microscopes apart from showing students enough for them to change mag, focus, and take a picture. Since I can't do everything, I've decided that more students will find basic light microscopy more generally useful in the future than EM techniques. However, the students really love using the "big iron" and are especially enthusiastic about SEM.
Cutting thin sections with glass knives has also been a bottleneck for my students so I was glad to see the discussion on this recently. Thanks, Dick, for summarizing the responses. -- Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
We took one of the smaller negative holders for which we didn't foresee a need and submitted it along with a TEM negative to a machinist in our shop. He was able to mill the opening such that it was the ideal size for the 3.25 x 4.25 negative. To give it that professional look he completed the job by touching up the milled edges with black paint.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Paula Allan-Wojtas [mailto:AllanWojtasP-at-em.agr.ca] Sent: Thursday, April 26, 2001 12:17 PM To: microscopy-at-sparc5.microscopy.com
Hi, all,
Following the recent discussion thread of scanners, I wanted to say that we are the proud owners of a Polaroid SprintScan 45 Ultra, and are very happy with the results we have gotten from it. It came with a number of negative holders covering a range of sizes from 35mm to 4x5, except one for 3.25x4.25 TEM sheet film. Right now we're trying to think of ways around this problem and would be happy to hear offline from anyone who has some ideas and/or solutions. I would even entertain the idea of having a holder made (this is not an option locally for me).
Thanks in advance for any help (and there is usually plenty, thanks to this list!).
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Tel: (902) 679-5566 FAX: (902) 679-2311
email: allanwojtasp-at-em.agr.ca
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Hello, I feel that I must add to this because as a general statement I believe it is mis-leading ... with respect to carbon black & many other carbon species... some 7000 images of carbon later I suggest that watching carbon morph in the beam is an exception not the rule. If you sit on anything long enough it will collect carbon but let's remain objective. Among other things, I have looked at CBs right out of reactors & extracted from polymers. How clean your instrument is can be a significant issue. My tool is a JEOL 2010, LaB6 operating at ~2x10-5Pa, & I use an LN2 cooled anti contamination device, ACD in Jeol lingo. The stage is room temperature and I usually work at 100 kV for the contrast enhancement. I have witnessed such things in experimental materials that I will term "highly reactive carbons". This lore of morphing carbon may be the most commonly used challenge of conclusions drawn from TEM data. Stories of exceptions tend to propagate over space & time. Don't get me wrong, it does happen.
Diverging a bit...
Of much more concern to anyone working in carbon is the carbon contamination supplied on new, right out of the box carbon coated grid formvar &/or Lacey carbon grid. The is a real problem for the manufactures & to my knowledge, despite any claims you may hear, no one has solved this problem. Once you understand & know the many habits carbon can have, that is the stuff supplied on new grids, you will realize that many images you see in the literature could easily be store bought carbon grid contamination. I have personally recorded dozens of such images... onions, fibers, rods, angular faceted bits, plates, turbostratic plates, polycrystalline graphite, CB looking stuff, graphitic ribbons & oh the nano structures, horns, fibers, .... Oh yea, watch out for very thin, low contrast crystals that throw a textbook perfect [001] hexagonal DP. it is not carbon. I don't know what it is. Weather on not you have realized it, you have seen them in imaging mode & probably passed them off as film thickness irregularities. I am looking at an image computer directory titled "grid stuff". it contains 12 images recorded on one new out of the box lacey carbon grid. Their titles are: hex plate, nest-2, nest -1, plates, rapping ribbon, plates plus, ribbon, fiber, rodlike, nanohorn, elongated hex. To all of this, let me refer you to an article recently or soon to be published in Carbon. I believe the author's name is Harris. I apologize for the poor quality reference, I've spaced out my copy of the preprint. It pretty much reads as something I have contemplated writing over the years but...
it's real folks.
Bruce Brinson Rice U.
Guess there is a reason ASTM standards require 1000 representative images.
Chuck Butterick wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Carbon black in polymer or rubber will cause no more problems than } most any other hydrocarbon. The electron beam does sublimate the } plastic. Especially when working with pure carbon black, one can } essentially watch a contaminant layer grow on the constituent } particles of a given aggregate. A cold finger is a very good idea but } not absolutely required if you are just scanning the sample. } } Our Philips 300 is still in excellent operating condition after 28 } years of carbon black work. } } Chuck Butterick } Engineered Carbons, Inc. } Borger, TX } } ______________________________ Reply Separator _________________________________ } Subject: carbon black in polymer } Author: Margaret Miller {MILLERMM-at-uthscsa.edu} at INTERNET-MAIL } Date: 4/26/01 2:27 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does carbon black cause a problem in the TEM? What percautions should I } take? Is the use of the cold finger recommended? } }
Just turn the So-163 film sideways on the 4x5 holder.
Paul PPG Industries
-----Original Message----- } From: Gerroir, Paul J [mailto:Paul.Gerroir-at-crt.xerox.com] Sent: Friday, April 27, 2001 6:28 AM To: Paula Allan-Wojtas; microscopy-at-sparc5.microscopy.com
Paula,
We took one of the smaller negative holders for which we didn't foresee a need and submitted it along with a TEM negative to a machinist in our shop. He was able to mill the opening such that it was the ideal size for the 3.25 x 4.25 negative. To give it that professional look he completed the job by touching up the milled edges with black paint.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Paula Allan-Wojtas [mailto:AllanWojtasP-at-em.agr.ca] Sent: Thursday, April 26, 2001 12:17 PM To: microscopy-at-sparc5.microscopy.com
Hi, all,
Following the recent discussion thread of scanners, I wanted to say that we are the proud owners of a Polaroid SprintScan 45 Ultra, and are very happy with the results we have gotten from it. It came with a number of negative holders covering a range of sizes from 35mm to 4x5, except one for 3.25x4.25 TEM sheet film. Right now we're trying to think of ways around this problem and would be happy to hear offline from anyone who has some ideas and/or solutions. I would even entertain the idea of having a holder made (this is not an option locally for me).
Thanks in advance for any help (and there is usually plenty, thanks to this list!).
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Tel: (902) 679-5566 FAX: (902) 679-2311
email: allanwojtasp-at-em.agr.ca
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
I used to teach an EM course to the graduate students here. It would run for 10 weeks and would consist of a 2-2.5 hour lecture period followed by a lunch break and then a 4 hour lab session during which I would teach them the "technique du jour" (fixation, embedding, block trimming, thick sectioning, thin sectioning, SEM sample prep, darkroom techniques, use of the EM....and in my really crazy days, alignment of the column!). the class would then be expected to come in during the week to put into practice what we had done in class. by the end of the class I expected each to have produces block, slides with stained thicks, grids with thins and 8X10 printed micrographs that showed me more than highly enlarged organelles.
I was also supposed to be running the dept's EM facility through all this. I would tell all of my usual clients not to expect anything substantial from me while the course ran, because I was too busy holding the hands of my 5-10 students. It would amount to 20-30 contact hours/week for the 10 week run. Exhausting.
Now that my facility is a college core, I can't just "drop out" for 10 weeks, and so the course is no longer offered.
Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Deb and others,
An interesting thread...
You make mention of "the EM staff" - I'm wondering how many people are actually involved in teaching the course components, and how much time do they collectively spend, for example per student per week??
I'm curious to know because we have two people involved in teaching a full semester Bio EM course for up to 8 students. The students in the end turn in a fairly polished portfolio (with about a dozen 8x10 micrographs plus figure legends, from three different tissues that they have processed, cut, and scoped).
Questions have recently arisen as to how many man-hours we 'should' allot to such a course, which is far outside the norm for our other lab courses.
I'm interested in off-line dialogue, if others are.
Ann Hein Lehman EM Facility Manager Trinity College Hartford CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu
-----Original Message----- } From: Dr Deborah Stenzel [mailto:d.stenzel-at-qut.edu.au] Sent: Friday, April 27, 2001 2:08 AM To: Microscopy-at-sparc5.microscopy.com
Dear Robert and all
I have the pleasure of teaching full semester (13 week) EM course to undergraduates at Queensland University of Technology in Brisbane, Australia. We've offered this course for quite a number of years - in fact, even had it extended from part of a broader topic to a full semester just for EM! The course is offered as part of the School of Life Sciences undergraduate program and, hence, primarily addresses biological EM. However, it is open to students from other disciplines - presently we do not have a full EM course in the physical sciences, although my colleague Dr Thor Bostrom teaches several weeks of EM in various courses for physics, chemistry and engineering undergraduate students.
Apart from this, I will usually run single 2 hour practical sessions for a couple of other undergraduate courses (mainly microbiology, where students look at negatively stained microbes they have isolated from various sources). I'll often be asked to give a half hour (sometimes an hour) introductory EM lecture to these students, prior to the practicals.
I also teach a brief introduction (2-3 weeks) to EM as part of a post-graduate course on cell structure and function in pathology. Other post-graduates needing EM as part of their research are taught individually by staff in our EM Facility, to the level that their projects require.
My full semester EM course is offered in the third year of the B. Applied Science course. This year I have 30 students - the numbers have been increasing over the past few years from a previous average of 20. I usually also have a couple of post-graduate (research) students attending the lectures - these students will be using our EM facilities as part of their research, and I find this is an effective means of giving them some useful background and theory to support their laboratory work.
For this course, I am allocated two by one hour lectures each week (total of 26 hours lectures for the semester). I cover basic instrumentation (TEM, SEM, preparation equipment) and biological sample preparation methods, including cryotechniques. I also include immunolabelling methods, some cytochemistry, microanalysis (Thor Bostrom gives these lectures), a few interesting "non-biological" applications, and a brief introduction to some other imaging methods (confocal, AFM etc).
Practical sessions have been a bit of a "challenge" as student numbers have increased and EM staff numbers have decreased! However, I've now got a system that works, and doesn't cause me (or the equipment) too much stress. I have two by 3 hours practical sessions running each week - ie. 6 hours of my teaching time per week (total of 72 hours of my teaching time for the semester).
Each student attends a total of 24 hours practical sessions for the semester (this fits with our "official" allocation for total practical time for a course of this credit point value). I divide the class into small groups - 3 to 4 students per group, with current student numbers. For preparation lab based pracs (where we have enough space and the students don't need such close supervision), I'll have a number of these small groups attending the same practical session. For EM operation and viewing pracs, I'll only have one small group attending at a time (usually 1 or 1.5 hours of the session for each small group). This has enabled me to run practical sessions almost single-handed - money to employ tutors/demonstrators or other teaching assistants has been a bit difficult to obtain! If anyone would like further information on the practicals I run or on the logistics of this, please contact me by direct email - does anyone else out there run EM practicals for undergraduates??? I haven't seen a rush of emails in response to Robert's questions.
Fortunately, my university is still keen to have students doing "hands-on" practicals in their courses. However, I've certainly had to decrease practical work substantially since I first started teaching in this EM course. A lot of this is due to decreased staff numbers in our EM Facility - a problem that I guess most of us face. I teach other courses (microbiology, parasitology) and encounter the same problems there - it is now very difficult to get competent, experienced people to assist in practical classes, particularly for more advanced or specialised courses.
On the bright side, I've found the students enjoy EM a great deal - and it is one of the few opportunities our undergraduates get to use expensive and (relatively!) modern instrumentation, even if it is only on a very limited scale. I've not had many students who don't show a great deal of enthusiasm when told it is now their turn to operate the TEM or SEM and take some micrographs. This semester, more so than previously, at the end of each practical session quite a number of my students have taken the time to thank me for running the practicals (even after the night classes which finish at 8pm). I figure that has got to be a good sign!!!
Regards Deb ***************************************************** Dr Deborah Stenzel Lecturer (Microbiology) School of Life Sciences and Applications Specialist (Biological) Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434 Brisbane 4001 Australia
We are a diagnostic EM lab in surgical pathology, using wet photography processes. If we receive a sample early in the morning, we will have labeled prints in the pathologists hands before 4:30pm the same day. It is basically 8 working hours for most rush cases. If we get the sample later in the day, we will finish the next morning, and prefer to polymerize overnight at 70 deg. C for epon araldite.
In order to speed things up, we put plastic in over at 100 deg C for 1 hr 15 min. The plastic isn't as good, but it does the job. We cure it for 5 minutes.
We offer two EM courses here, one SEM and one TEM. Each is an elective course taken by a mixture of seniors and graduate students, which runs once a year for a full semester. I teach the TEM course, and enrollment is generally anywhere from six to ten students. (More than ten would be a problem in lab.) We have two 50-minute class periods a week, to give the students some foundation in relation to the instrument and electron optics, specimen prep., electron diffraction (we're a materials department), and imaging. There is also a two-and-a-half hour lab session each week, which covers basic operation of the microscope, an introduction to specimen preparation, and indexing of diffraction patterns.
Gill
Dr. Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech Socorro, NM 87801 (505) 835-5653 gbond-at-nmt.edu
----- Original Message ----- } From: "Robert Fitton" {fittonro-at-luther.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 26, 2001 2:40 PM
Hello fellow microscopists:
Does anyone know good protocols for-
1. retina
2. colagen (to prevent shrinkage in skin tissue)
Thanks again, Tim Quinn University of Kansas Museum of Natural History
We have a range of interactive EM courses set out in e-book form. We cover basic SEM, TEM and EDX with more advanced courses in SEM. The pictorial data are in the form of bitmaps and we allow purchasers to use these in their own presentations if they wish.
Full details are on our web site
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
Margaret, I take it from your 'subject' you have a solid block of polymer with a CB dispersion. If this is correct you will have no problem looking at a thin section. Contrast will be low but a low KV ~60 would be good provided you keep the sections thin. For better contrast you can pyrolyze the polymer to improve the contrast. If you would like to do this let me know and I fill you in on the details. Russ Gillmeister Microscopy Xerox Corp. RGillmeister-at-crt.xerox.com
-----Original Message----- } From: Margaret Miller [mailto:MILLERMM-at-uthscsa.edu] Sent: Thursday, April 26, 2001 3:28:PM To: MSA
Does carbon black cause a problem in the TEM? What percautions should I take? Is the use of the cold finger recommended?
Hi! I have a problem with my EDS system. I bought it in 1992 with a SEM. Now, the resolution and the quantification results are getting worse. Maybe it should be enough changing the window and the detector and testing all. I'd like to have more than one offering about repairing manufactory but I don't know to ask about it . Could anyone help me? Thank's in advance.
I also am in a very similar situation, with a detector of about the same age, which appears to have slowly but steadily lost resolution (we're up to nearly 200 eV full width at half max on the Mn K-alpha line).
I have asked the manufacturer how and why this kind of degradation occurs, but haven't gotten a clear answer. I realize there are multiple components of the system from which the problem could be coming (it manifests itself as an approximately constant 50 eV additional fwhm component on all peaks in the spectrum regardless of at which energy). However, I'd like to know if there are reasons why this might be unavoidable in a detector of this age? If not, what are possible causes and how can one prevent it from happening?
Thanks, Wharton
} -----Original Message----- } From: Romina Belli [SMTP:belli-at-science.unitn.it] } Sent: Monday, April 30, 2001 8:04 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM-EDS: change or repair? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi! } I have a problem with my EDS system. I bought it in 1992 with a SEM. Now, } the resolution and the quantification results are getting worse. } Maybe it should be enough changing the window and the detector and testing } all. I'd like to have more than one offering about repairing manufactory } but I don't know to ask about it . Could anyone help me? } Thank's in advance. } } Romina Belli
Does anyone have a good protocol for retina fixation for TEM?
I'm dealing with specimens collected in the field and it appears that the initial field fix- "cacodylate and glut" did not preserve the "rods and cones".
A good recomendation for a field fix would be appreciated. Would Karnovsky's work?
I also need to fix frog skin tissue, causing minimal shrinkage. Any suggestions?
I teach three EM classes here which are undergrad and grad level: EM Theory (2 credits Spring Semester), TEM Lab (3 credits Spring semester), and SEM Lab (2 credits fall semester).
The EM Theory class meets 2x 50 mins each week and is lecture only (I bring in lots of hands on show and tells). It covers SEM, TEM, LM, photography (Silver & digital), sample prep, and AEM. I have a number of students who take this without taking the lab courses. The Lab courses require the EM Theory class.
The both labs meet as a group (class limit of 8-9 students) for 2-5 hours on Mondays (Scheduled for 3 hours but fixation days run long). The students then meet one-on-one with a TA for 2hrs on the scope each week. The students “drive” and the TA’s train/assist/watch. Both Labs cover full microscope operation (including alignment, operating parameters, imaging modes, and photography), sample prep (from wet to scope, including embedment, ultramicrotomy, CPD, particulates, staining techniques - 3 separate preps for TEM and 5 for SEM), darkroom, and digital publication plate production. Students take a written exam, an oral exam on the scope (once they “pass” the scope exam they can use the scope without supervision), and turn in a pretty polished portfolio of images. TEM students spend significant amounts of additional time sectioning, staining, and developing film and contact printing.
Yes, these labs are labor intensive (teaching and students), but by the end of either course the goal is that they are qualified TEM or SEM users. Most of the students continue on utilizing the scope for research (most undergrads taking the actually have formal research projects with faculty members that they continue for 1-2 years plus summers, or on to a masters degree). Students are accepted into the lab classes based on research needs.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
At Chicago State University I teach a 4 credit hour course in TEM in the fall each year. The students have 2 one-hour lectures per week and 10 hours of supervised lab experience per week. I have usually have 4 to 8 students each year. Since students are scheduled into the lab 2 at a time, this means that someone is with students 30-50 hours per week. For the last few years I have had a technologist assistant who has been able to work with the students, so that we have been able to continue with research and other work during the semester. It really does take two people for this sort of class. In the spring I have taught a 3-credit hour class in SEM. This one has 2 hours of lecture and 6 hours of supervised lab per week. When my students are done with the class, they can prepare specimens competently, use the TEM or SEM to take pictures, so some alignment, understand the principles, and present a simple research project to the department. In the past that was done as a poster or with slides, but in the last few years they have been preparing Power Point presentations.
I teach three EM classes here which are undergrad and grad level: EM Theory (2 credits Spring Semester), TEM Lab (3 credits Spring semester), and SEM Lab (2 credits fall semester).
The EM Theory class meets 2x 50 mins each week and is lecture only (I bring in lots of hands on show and tells). It covers SEM, TEM, LM, photography (Silver & digital), sample prep, and AEM. I have a number of students who take this without taking the lab courses. The Lab courses require the EM Theory class.
The both labs meet as a group (class limit of 8-9 students) for 2-5 hours on Mondays (Scheduled for 3 hours but fixation days run long). The students then meet one-on-one with a TA for 2hrs on the scope each week. The students “drive” and the TA’s train/assist/watch. Both Labs cover full microscope operation (including alignment, operating parameters, imaging modes, and photography), sample prep (from wet to scope, including embedment, ultramicrotomy, CPD, particulates, staining techniques - 3 separate preps for TEM and 5 for SEM), darkroom, and digital publication plate production. Students take a written exam, an oral exam on the scope (once they “pass” the scope exam they can use the scope without supervision), and turn in a pretty polished portfolio of images. TEM students spend significant amounts of additional time sectioning, staining, and developing film and contact printing.
Yes, these labs are labor intensive (teaching and students), but by the end of either course the goal is that they are qualified TEM or SEM users. Most of the students continue on utilizing the scope for research (most undergrads taking the actually have formal research projects with faculty members that they continue for 1-2 years plus summers, or on to a masters degree). Students are accepted into the lab classes based on research needs.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
The Microscopy and Imaging Facility of the University of Calgary requires an Electron Microscopy Technician to fill a newly created position within the unit.
The Microscopy and Imaging Facility is a service resource for the University of Calgary supporting users campus wide. The unit works in all areas of science including medicine, biology, chemistry, materials engineering, and geology. The unit is equipped with six electron beam instruments, X-ray micro analyzers, confocal microscope, digital imaging / image analysis and support resources. It is the largest unit of its type in Western Canada.
The successful applicant will have work experience in the operation and routine maintenance of transmission electron microscopes. The demonstrated ability to teach and instruct new users on instrumentation will be a definite asset. Competence in the operation and use of computers and common software packages is a must.
Salary for this position will be dependant on education and experience in the field.
Please send by May 14, 2001 your cv and a covering letter that includes details of your experience that is relevant to this position to: Dr. John Reynolds Department of Cell Biology and Anatomy University of Calgary, 3330 Hospital Dr NW, T2N 1N4, Calgary, Alberta, Canada.
I also am in a very similar situation, with a detector of about the same age, which appears to have slowly but steadily lost resolution (we're up to nearly 200 eV full width at half max on the Mn K-alpha line).
I have asked the manufacturer how and why this kind of degradation occurs, but haven't gotten a clear answer. I realize there are multiple components of the system from which the problem could be coming (it manifests itself as an approximately constant 50 eV additional fwhm component on all peaks in the spectrum regardless of at which energy). However, I'd like to know if there are reasons why this might be unavoidable in a detector of this age? If not, what are possible causes and how can one prevent it from happening?
Wharton
} Hi! } I have a problem with my EDS system. I bought it in 1992 with a SEM. Now, } the resolution and the quantification results are getting worse. } Maybe it should be enough changing the window and the detector and testing } all. I'd like to have more than one offering about repairing manufactory } but I don't know to ask about it . Could anyone help me? } Thank's in advance. } } Romina Belli
Dear Wharton and Romina, Our Kevex detector was installed in the early 80s, and every so often we have had to warm up and pump out the detector to get back the specified resolution (145 eV). Last year, when this didn't work, we sent the detector to Doug Connors (tnas1-at-msn.com), and he cleaned and overhauled it for a good price and got 147 eV resolution. I have no connection to Doug other than as a satisfied customer. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
This is a question about the FEI FIB 200: We are interested in converting our Pt Gas Injection System into Au, but have been told about possible negative effects on the instrument.
Any data/experience/thoughts on Au deposition using FIB (any model) and its effect on the instrument performance will be greatly appreciated!
Thanks! P.S. I realize it is not quite a microscopy question. I have posted it to the FIB-users list, but received only one response so far. Apologize to those who will get this message for the second time.
******************************** Katharine Dovidenko, Ph.D. Scientist UAlbany Institute for Materials and Center for Advanced Thin Film Technology University at Albany SUNY www.albany.edu/cat
251 Fuller Rd. Albany, NY 12203 USA Phone: (518) 437-8781 Fax: (518) 437-8687
} -----Original Message----- } From: Gonzalez-Cabezas, Carlos } Sent: Monday, April 30, 2001 2:37 PM } To: Goheen, Michael P. } Subject: RE: SEM position } } I was asked to post this job opening on the listserver. } } Mike Goheen } } SEM/EPMA tech wanted. The Indiana University School of Dentistry is } looking for a technician for its new digital electron microscopy } laboratory. } A JEOL LV-5310 scanning electron microscope and JEOL 8900R electron probe } microanalyzer are in the process of being installed in a renovated lab in } the IU School of Dentistry. The electron microscopes will have energy- and } wavelength-dispersive spectrometers and are fully digitized. The } laboratory } will serve the research and teaching interests of several units on campus } in } addition to Dentistry including the IU School of Medicine, and the Purdue } School of Science (Departments of Geology, Biology, Chemistry, and } Physics) } and Purdue School of Engineering at Indianapolis. We seek a candidate who } has a range of interests in spatial variations of the microstructure and } composition of materials, and skills in one or more fields such as } computer } technology, electron microscopy, materials science, engineering } technology, } biomedical and geological research. Please contact Dr. Carlos Gonzalez, } Preventive Dentistry Department, IU School of Dentistry. } } Dr. Carlos González-Cabezas, DDS, PhD } Director of the Confocal & Scanning Electron Microscopy Facility } Indiana University School of Dentistry } CGONZALE-at-IUPUI.EDU } } } } -----Original Message----- } From: Goheen, Michael P. } Sent: Wednesday, April 11, 2001 2:17 PM } To: Gonzalez-Cabezas, Carlos } Subject: RE: SEM position } } } -----Original Message----- } From: Gonzalez-Cabezas, Carlos } Sent: Wednesday, April 11, 2001 12:20 PM } To: Goheen, Michael P. } Subject: SEM position } }
{bold} {color} {param} ffff,0000,0000 {/param} {bigger} FYI: This is the last day for early registration for the FRET/FLIM Symposium in San Antonio.=20 Abstracts will be accepted until June 1st.
{/bigger} {/color} {/bold} {bigger} The University of Texas Health Science Center will host a symposium sponsored by Hamamatsu Photonics KK on
{paraindent} {param} left,out {/param} "Spatial resolution of early signalling processes in cells"=20
{/paraindent} Christoph Biskup,=20
{paraindent} {param} left {/param} "Fluorescence lifetime and energy transfer measurements in living cells with a confocal laser scanning microscope and a streak camera"=20
{/paraindent} Robert Clegg,=20
{paraindent} {param} left {/param} "Real-time fluorescence lifetime-resolved imaging - why we do it, how it's done, and applications for biology and medicine."=20
{/paraindent} Michael Edidin
{paraindent} {param} left {/param} "Photobleaching FRET microscopy: practice and theory"
{/paraindent} Enrico Gratton =20
Hans Gerritsen
{paraindent} {param} left {/param} "Fast fluorescence lifetime imaging"=20
{/paraindent} Jesus Gonzalez
{paraindent} {param} left {/param} "FRET Probes and Assays for Drug Discovery"=20
{/paraindent} Brian Herman
{paraindent} {param} left {/param} "FRETing over the apoptotic cascade"
{/paraindent} Thomas Jovin
{paraindent} {param} left {/param} "Extending the capabilities of FRET and FLIM for molecular and cellular biology: phFRET (photochromic FRET), rFLIM (anisotropy FLIM), spectrally-resolved and optical-sectioning FLIM"
{/paraindent} Karsten K=F6nig
{paraindent} {param} left {/param} "Multiphoton microscopy with submicron spatial and picosecond temporal resolution"=20
{/paraindent} Wen-hong Li
{paraindent} {param} left {/param} "Towards the development of highly luminescent lanthanide complexes for FRET and FLIM"=20
{/paraindent} Paloma Mas (substituting for Steve Kay)
{paraindent} {param} left {/param} "Functional interaction of phytocrome B and cryptochrome 2"=20
{/paraindent} Atsushi Miyawaki
{paraindent} {param} left {/param} "Imaging of cellular functions by FREting"
{/paraindent} Ammasi Periasamy
{paraindent} {param} left {/param} "Quantitation of Protein Signals in a Single Living Cell: Wide-field, Confocal, Two-photon and Lifetime FRET Microscopy"=20
{/paraindent} Alexander Sorkin
{paraindent} {param} left {/param} "Interactions of the EGF receptor with adapter proteins during endocytosis" =20
{/paraindent} Roger Tsien=20
{paraindent} {param} left {/param} "FRET based readouts of intracellular messengers and protein interactions"=20
Dear Wharton and Romina, I am running two EDX detectors that are older than Romina's, one 1985 and one of similar age that I bought used. Both still meet their original spec of 149 and 146 eV at Mn Ka. When I have had a degradation of resolution, I turned off the bias, grounded it out with a paper clip on the detector bias connector and warmed up the detector until all the frost was gone from inside the dewar. I used warmed air from a hair drier, blown into a hose down to the bottom, but there other methods that can be used. When the dewar was completely dry, I refillled with liquid nitrogen, let it cool overnight and applied bias the next day. The bias should be on at least one hour before testing the resolution. In one case this cured the resolution, but degraded the holding time for liquid nitrogen, so I then had the dewar re-pumped. I would recommend that step, at least, before buying a new detector. There are also EDX detector repair companies that will diagnose and repair detectors for less than the cost of a new one. I have also had a grounding problem that gave high noise on the detector, because the case on the pre-amp oxidized. A little emery paper cured that. That showed more high dead-time than degraded resolution. At 08:58 AM 4/30/01 -0500, you wrote: } } Dear List, } } I also am in a very similar situation, with a detector of about the same } age, which appears to have slowly but steadily lost resolution (we're up to } nearly 200 eV full width at half max on the Mn K-alpha line). } } I have asked the manufacturer how and why this kind of degradation occurs, } but haven't gotten a clear answer. I realize there are multiple components } of the system from which the problem could be coming (it manifests itself as } an approximately constant 50 eV additional fwhm component on all peaks in } the spectrum regardless of at which energy). However, I'd like to know if } there are reasons why this might be unavoidable in a detector of this age? } If not, what are possible causes and how can one prevent it from happening? } } } Thanks, } Wharton } } } -----Original Message----- } } From: Romina Belli [SMTP:belli-at-science.unitn.it] } } Sent: Monday, April 30, 2001 8:04 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: SEM-EDS: change or repair? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi! } } I have a problem with my EDS system. I bought it in 1992 with a SEM. Now, } } the resolution and the quantification results are getting worse. } } Maybe it should be enough changing the window and the detector and testing } } all. I'd like to have more than one offering about repairing manufactory } } but I don't know to ask about it . Could anyone help me? } } Thank's in advance. } } } } Romina Belli } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Job Posting submitted by Dr. David P. Bazett-Jones
Service Manager, Electron Microscopy Facility
Date Posted: April 30, 2001
Position Status: Full-time, Fixed term
Department: Cell Biology Research Institute
Available: August 1, 2001
Description of the Position: You will share responsibility for the operation and maintenance of transmission and scanning electron microscopes in a new Bioimaging Facility co-sponsored by teaching hospitals in the University of Toronto. The microscopes include an ESEM (FEI/Philips) and a 200 kV TEM (FEI/Philips) equipped with EDX, GIF and cryostage. You will also be responsible for coordination and management
of electron bioimaging services required by investigators of the Hospital for Sick Children Research Institute.
Qualifications: As an ideal candidate, you have completed a M.Sc. in biological sciences, or have completed a B.Sc. with experience in analytical electron microscopy, ultramicrotomy and other sample preparation techniques. Strong computer skills are an asset.
You possess excellent verbal communication and organizational skills. You have the ability to work well independently and in a team.
Hours : 35 hours/week
Salary: $39,848.95 - $50,277.67
Available to: Internal and External Candidates
Deadline: May 9, 2001
Submit Resume to : Erin O’Hare The Hospital for Sick Children 555 University Avenue, Toronto, Ontario M5G1X8
Mananged to get most of the OS updated done this evening. There may be a few hiccups over the next couple of days so be patient. Be prepared for the occasional error message while I reset and fine tune all the system parameters
Cheers.... Nestor Your Friendly Neighborhood SysOp
Dear All Being a botanist and all, I know the cube root of very little about this really, but my understanding is that gold atoms can diffuse into and "poison" semiconductors, and that gold should never be used for specimen coating etc. in any SEM / FIB which may be used to examine or test semiconductors where the semiconductor functionality must be maintained. A) Is this relevant to Katharine's question? B) Is it true or an urban myth?
Chris
----- Original Message ----- } From: "Katharine Dovidenko" {KDovidenko-at-uamail.albany.edu} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, April 30, 2001 6:57 PM
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Wharton, It may be as simple as cleaning out your dewar and or outgassing your window. We were losing resolution on our EDAX detector. It was brought up to room temperature for a day or so and the resolution improved greatly. It worth a try if the manufacturer allows it. Clean out any debris from the dewar and make sure your detector isn't powered up during the warm up. Good Luck, Russ Gillmeister Microscopy Xerox Corp. RGillmeister-at-crt.xerox.com
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Monday, April 30, 2001 9:59:AM To: 'Romina Belli'; Microscopy-at-sparc5.microscopy.com
Dear List,
I also am in a very similar situation, with a detector of about the same age, which appears to have slowly but steadily lost resolution (we're up to nearly 200 eV full width at half max on the Mn K-alpha line).
I have asked the manufacturer how and why this kind of degradation occurs, but haven't gotten a clear answer. I realize there are multiple components of the system from which the problem could be coming (it manifests itself as an approximately constant 50 eV additional fwhm component on all peaks in the spectrum regardless of at which energy). However, I'd like to know if there are reasons why this might be unavoidable in a detector of this age? If not, what are possible causes and how can one prevent it from happening?
Thanks, Wharton
} -----Original Message----- } From: Romina Belli [SMTP:belli-at-science.unitn.it] } Sent: Monday, April 30, 2001 8:04 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM-EDS: change or repair? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi! } I have a problem with my EDS system. I bought it in 1992 with a SEM. Now, } the resolution and the quantification results are getting worse. } Maybe it should be enough changing the window and the detector and testing } all. I'd like to have more than one offering about repairing manufactory } but I don't know to ask about it . Could anyone help me? } Thank's in advance. } } Romina Belli
We are in the process of updating the Microbeam Analysis Society's membership database. If there are any changes in your personal information (address, phone/fax #'s, email, etc.) as listed in the 2000 directory and you have not made the necessary changes on your 2001 renewal form, please email me with the updated information. Although we will not be printing a new directory this year we would like to keep everyone as current as possible with the membership information.
If you are not a member of MAS and would like to join, please contact me for more information and an application form.
Thanks, Lou Ross MAS Membership Services PMB #141 2101 W. Broadway Columbia, MO 65203-1261 (800) 4MASMEM url: www.microanalysis.org
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} We are currently looking for a used ultrcut microtome in good to excellent {BR} condition. {BR} John Hoffpauir {BR} Cooper hospital {BR} Camden NJ {BR} 08107 {BR} 856 757-7781 {/FONT} {/HTML}
Just a general question for the List - are those 10mm x 10mm cylindrical copper (or Al) Jeol type stubs still in wide usage in the SEM community? Do newer Jeol machines still use them? The reason I'm asking is that we have a couple cabinets full of these old stubs from when we used to have a Jeol back in the mid-70's, but of course can't look at them now with our current instrument. I'd like to toss them so we can modify the cabinets to accept our pin-type stubs, and I'd feel better about doing so if I thought it would be hard to find an instrument that could still look at these old ones. Of course, if it turns out that I can't find documentation to indicate what's on these stubs, they'll be going in the bin anyway.
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
Workshop Announcement University of California at Santa Barbara
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 4-day workshop will be offered from August 20 through August 23, 2001 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be an intensive lecture/laboratory series that will enable participants to develop theoretical and hands-on expertise with light microscopes. Attendees will closely interact with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski differential interference contrast, and darkfield imaging will be taught and attendees will use microscopes equipped to perform these optical enhancement techniques. In addition, the theory and practice of electronic image acquisition (analog and digital) will be discussed and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
The Nikon 990 has a pretty good macro mode and I have found a supplier of a macro lens system that is excellent, but I need a stand that has rough height adjustment like you would find on a stereo scope. Has anyone found a good solution for this presuming that the sample would sit on a table or stand and the Nikon lens would be lowered to the desired height. I know this is typically done with a copy stand, but I find them over-kill (large and provides own illumination). I would use the illumination from my stereo scope so all I need is a small mechanical stage.
Being an electronics technician, I only know enough about optics to be dangerous, but use microscopes often. I am trying to understand what we have, in order to figure out what else could help us out. I would appreciate information for a "layman", or pointers web sites that might enlighten me.
First, I need help understanding the markings on our objective lenses. The question marks indicate what I don't even have an inkling of the meaning.
#1: Zeiss (The maker of our LSM & this lens) Epiplan-NEOFLUAR ( ? ) 100x/0,90 (magnification/numerical aperture) 44 23 80 ( ? )
#3 research devices (The maker) infrared (illumination designed for) Trans 100 IRN ( ? ; mag ; near IR ? ) 0.90 ( NA )
Is it possible to calculate the working distance from the available information?
When a lens is made for infrared use, what is different about it? Are the lens coatings different, or totally absent? Is the lens glass special?
If you read this far, thank you! I thought I would find the answers to these questions, and many more, on an episode of "Soap," but it didn't work. The list is a wonderful pool of knowledge, so don't fail me now!
Chris: if you MUST maintain functionality, it's best not to coat at all. My next choice would be carbon coating, which can be removed by ashing in an O2 plasma. The 3rd choice is Au/Pd, which can be removed with a wet etch of iodine/potassium iodide, but this can attack any exposed aluminum metal on the die.
The biggest problem with using a coating on semiconductors one wishes to remain functional is getting all the coating off to prevent shorts/leakages between the device pins. I don't think it's relevant to Katherine's question as she is concerned about negative effects to her FIB, not the devices.
Most semiconductors have a passivation layer (usually silicon nitride about 1 micron thick) over their surfaces and this protects from unwanted contaminants. I'm no device physicist, but I think the device would have to be heated to a high temperature for any gold implanted in the top atomic layers to diffuse into the active junctions and cause trouble. You want to keep Au out of the fab, but after the passivation is deposited and is intact, the device is fairly impervious to metals sputtered on top. Some older devices used to have Au plated on their backs to help attach them in their packages.
I've used gold (sputter and evaporative) coating over the years to coat semiconductors for SEM exam, and haven't seen any adverse effects. Most of my problems were related to trying to get the coating off afterwards.
Chris Jeffree wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } Being a botanist and all, I know the cube root of very little about } this really, but my understanding is that gold atoms can diffuse into } and "poison" semiconductors, and that gold should never be used for } specimen coating etc. in any SEM / FIB which may be used to examine or } test semiconductors where the semiconductor functionality must be } maintained. A) Is this relevant to Katharine's question? B) Is it } true or an urban myth? } } Chris } } ----- Original Message ----- } } From: "Katharine Dovidenko" {KDovidenko-at-uamail.albany.edu} } To: "'Microscopy-at-MSA.Microscopy.Com'" } {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, April 30, 2001 6:57 PM } Subject: FIB: Au deposition instead of Pt. } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } Dear all: } } } } This is a question about the FEI FIB 200: We are interested in } converting } } our Pt Gas Injection System into Au, but have been told about } possible } } negative effects on the instrument. } } } } Any data/experience/thoughts on Au deposition using FIB (any model) } and its } } effect on the instrument performance will be greatly appreciated! } } } } Thanks! } } P.S. I realize it is not quite a microscopy question. I have posted } it to } } the FIB-users list, but received only one response so far. Apologize } to } } those who will get this message for the second time. } } } } ******************************** } } Katharine Dovidenko, Ph.D. } } Scientist } } UAlbany Institute for Materials and Center for Advanced Thin Film } Technology } } University at Albany } } SUNY } } www.albany.edu/cat } } } } 251 Fuller Rd. } } Albany, NY 12203 } } USA } } Phone: (518) 437-8781 } } Fax: (518) 437-8687 } } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
For macro photography I use a machinist's height gauge. They are usually about 18" high with a smooth, but tight, sliding anvil that can
be machined to accept a camera adapter ring mount. A threaded fine adjustment allows for fine focus. Mounting a Nikon 990 will require that additional weight be added to the gauge's base to prevent the assembly from toppling. Lubricated glass plates and a tiny sandbox can
be used for orienting the sample relative to the camera lens.
Location: Eindhoven, Noord braband, The Netherlands
Question: L.S. I'm doing a survey on bone tissue under the microscope. I want to make the bone tissue visible from a large overview with a regular microscope to a small overview with a (E)SEM. Do you have any experience with this kind of survey's. What kind of problems can I run into (e.g. How can I mark the piece of bone so that I always will see the same spot of bone under the different microscopes?). Do you have reports of other students that have done a study alike this one. At forehand thanks for your time cheers koen kodde student medical engineering
We are doping plastics with high concentrations of dyes and would like to determine the optical transmission properties of these plastics in the wavelength range 350-800nm as a function of thickness of the doped plastic. We do not have the capability to accurately cut thin sections of these plastics to perform these measurements. Ideally we would like to have sections cut at 0.25, 0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5 and 5.0 microns at for a series of dye concentrations in these plastics and have the sections mounted on glass slides. The sections should be free of holes, scratches or other defects. We can cast the plastic to whatever shape is needed but the other dimensions need to be at least 5x5mm.
Is there a service or contract lab out there that can do the sectioning. Interested parties should contact me to discuss further details.
Pleases do not sent to me SU ---------------------- Forwarded by Sumalee Uthaithavorn/BKK/BE/PHILIPS on 2001-05-02 09:43 ---------------------------
milesd-at-US.ibm.com on 2001-05-02 07:33:29 To: Microscopy-at-sparc5.microscopy.com-at-SMTP cc:
Hello All.
Being an electronics technician, I only know enough about optics to be dangerous, but use microscopes often. I am trying to understand what we have, in order to figure out what else could help us out. I would appreciate information for a "layman", or pointers web sites that might enlighten me.
First, I need help understanding the markings on our objective lenses. The question marks indicate what I don't even have an inkling of the meaning.
#1: Zeiss (The maker of our LSM & this lens) Epiplan-NEOFLUAR ( ? ) 100x/0,90 (magnification/numerical aperture) 44 23 80 ( ? )
#3 research devices (The maker) infrared (illumination designed for) Trans 100 IRN ( ? ; mag ; near IR ? ) 0.90 ( NA )
Is it possible to calculate the working distance from the available information?
When a lens is made for infrared use, what is different about it? Are the lens coatings different, or totally absent? Is the lens glass special?
If you read this far, thank you! I thought I would find the answers to these questions, and many more, on an episode of "Soap," but it didn't work. The list is a wonderful pool of knowledge, so don't fail me now!
Greetings to Prof Holland Cheng, Prof Alex Hyatt and all list users.
I worked in a multi-users laboratory and we have facilities equipped for cryo TEM. I have been asked by my colleague to post the following questions:
1. Is it necessary to chemically fixed virus-infected samples for cryo TEM? 2. What are the methods and precautions for cryo TEM of unfixed virus-infected cells? 2. How to dispose the LN2? 3. What are the differences between chemically fixed and unfixed virus-infected samples for cryo TEM?
Hi, I saw your post and thought you might be interested in this...
When you access the Internet, your computer keeps permanent hidden records of your activities! I recently tried EE and I was shocked at what it uncovered on my hard drive.....It actually frightened me. It showed all that I had been doing even though I had deleted it. My advice is to check it out NOW I found it at http://ee1.20m.com Regards, Harry
If the ratio in magnifications between steps is only about 3x, you should be able to easily locate common features between successive pictures. You should have some overlap in magnification between your regular microscope and your ESEM. If not, I am sure you should not have more than a 3-fold difference. We often take an overview picture with a macro camera of concrete samples before putting them into the SEM. That is usually stretching a bit because we are going from 1x for our overview to 20x in the SEM and it is much harder to locate the same areas over that big a jump.
Of course, if you could scribe some marks outside the bone, that can provide you with registration points.
Warren
At 05:44 PM 5/1/2001 -0500, you wrote:
} Email: A.K.Kodd-at-stud.tue.nl } Name: Koen Kodde } } Organization: Technical University of Eindhoven } } Education: Graduate College } } Location: Eindhoven, Noord braband, The Netherlands } } Question: L.S. } I'm doing a survey on bone tissue under the microscope. I want to make the } bone tissue visible from a large overview with a regular microscope to a } small overview with a (E)SEM. Do you have any experience with this kind of } survey's. What kind of problems can I run into (e.g. How can I mark the } piece of bone so that I always will see the same spot of bone under the } different microscopes?). Do you have reports of other students that have } done a study alike this one. } At forehand thanks for your time } cheers } koen kodde } student medical engineering
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
A meeting organised by the Royal Microscopical Society and supported by EMAG, FEI UK Ltd, JEOL (UK) Ltd.,LEO EM Inc., Gatan UK, TVIPS GmbH
Department of Materials, Oxford University
Tuesday 3 July 2001
The past three years have seen a remarkable growth in field-emission-gun transmission electron microscopy (FEGTEM). Following the pattern of the two previous events, this meeting will focus both on novel instrumentation developments and on applications to a wide range of problems in the life sciences and the physical materials sciences.
There will be a small number of invited speakers, including
Dr Steven Fuller (Oxford University) Dr Owen Saxton (Cambridge University) Dr Rik Brydson (Leeds University)
representing these aspects of the subject.
The majority of the programme will be devoted to contributed papers. Contributions on all aspects of FEGTEM are now being sought. Abstracts should be sent as soon as possible (and in any case not later than 31 May 2001) to john.hutchison-at-materials.ox.ac.uk, from whom further information may be obtained.
A downloadable registration form, are available at: http://www.rms.org.uk/current%20events.html#fegtem
Further details are available from the organisers: Dr John Hutchison, tel +44 (0)1865 273705, john.hutchison-at-materials.ox.ac.uk Dr Rafal Dunin-Borkowski, tel +44 (0)1223 334564, rafal.db-at-msm.cam.ac.uk
Note. This meeting is timed to precede immediately the one-day EFTEM meting to be held on 4 July 2001, also in Oxford.
Rafal Dunin-Borkowski Department of Materials Science University of Cambridge Pembroke Street Cambridge CB2 3QZ, UK Tel +44 1223 334564 rafal.db-at-msm.cam.ac.uk Fax +44 1223 334567 http://www-hrem.msm.cam.ac.uk/~rdb/
In a message dated 5/1/2001 3:59:27 PM Mountain Daylight Time, thomasf-at-AGC.BIO.NS.CA writes:
} Just a general question for the List - are those 10mm x 10mm cylindrical } copper (or Al) Jeol type stubs still in wide usage in the SEM community? Do } newer Jeol machines still use them?
The JEOL 5900, which is currently their main conventional model, uses a 3/8" x 3/8" (9.5 mm x 9.5 mm) stub. The older JEOL 840 and 6000 series typically use a 12.3 mm x 12.3 mm stub.
I quickly checked a few old catalogs and found the prices to range from about USD $15 to $100 for 100 of these types of Al stubs (the 9.5mm stubs seem to be about 1/2 the price of the larger size).
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Dinner Choices: (Indicate Choice when RSVP'ing) Red Snapper, Basque Style London Broil Grilled, Fresh Vegetables & Smoked Mozzarella with Herbs in Puff Pastry
RSVP: On-line at http://www.amplifyllc.com/go/ncss. Preferred method. (Can use credit cards.) Or, email steve.rabin-at-alza.com. Or, call Steve Rabin at 1-650-564-5315
PLEASE RSVP BY MONDAY APRIL 30th.
(Please let us know if you're coming for dinner, or just the talk for headcount purposes)
****************** ABSTRACT ******************
Magnetic resonance microscopy (MRM) has been employed to investigate solvent transport phenomena in glassy and rubbery macromolecular systems. Time-dependent proton or fluorine imaging of solvent uptake allows one to distinguish between two different transport mechanisms. An exponential solvent concentration profile observed in the case of rubbery networks is consistent with Fickian behavior. In glassy systems, however, where solvent
induces a glass-to-rubber phase transition of the network, an extremely sharp solvent front is observed which propagates through the specimen as a shock wave at constant velocity which is typical of anomalous transport. MRM analysis forms the basis of a model of anomalous transport in macromolecular solids which couples diffusion of solvent with kinetics of the phase transition of the network. Analysis of solvent transport kinetics, front velocities, molecular diffusion constants and localized chain
****************** DIRECTIONS TO DAVID'S RESTAURANT ******************
David's Restaurant, 5151 Stars and Stripes Drive (off Tasman Dr., Santa Clara CA, at the Santa Clara Golf club)
} From the East Bay: Take 680 or 880 to Highway 237 toward Mountain View. Stay on 237 until the Great America Parkway exit. Follow Great America Parkway to Tasman Dr, take a left on Tasman. Approximately 0.5 miles on the left, turn left onto Centennial Drive. David's will be in front of you.
} From San Jose Take 101 North to Great America Parkway. Take Great America Parkway to Tasman Drive, turn right on Tasman. Approximately 0.5 miles on the left, turn left onto Centennial Drive. David's will be in front of you.
} From the Peninsula Take 101 South to Highway 237 toward Milpitas. Exit at Great America Parkway. Follow Great America Parkway to Tasman Dr, take a left on Tasman. Approximately 0.5 miles on the left, turn left onto Centennial Drive. David's will be in front of you.
Dear List, I have noticed some recent inquiries about resolution changes with Energy Dispersive Spectrometers and also the reported solutions. There seems to be some misinformation about the use and maintenance of these detectors. These detectors are basically a closed vacuum chamber which over time can have water vapor and other gases enter the system through the entrance window in the case of Light Element Windows or general out gassing due to deterioration of o-ring seals etc. When water vapor is present in the system it will collect on the cold finger of the detector and as such will create an ice layer on the Si(Li) crystal since it is also at near LN2 temperature. When this occurs, resolution of the detector degrades. One of the solutions reported was to allow the detector to warm up to room temperature and clean the dewar. Unfortunately this will only allow the process to reoccur in a short period of time. What is necessary is a thorough pump-out and bake-out of the detector after replacing various vacuum seals. This can only be accomplished by experienced technical personnel with the appropriate vacuum pump station. Obviously one can return the detector to the original manufacturer or some other repair facility. My company, X-Ray Optics/AAT is one of these EDX repair facilities and if anyone is interested they may contact me through my web site at www.xraydetectors.com I hope this information is useful to the members of the listserver. Regards, Jim Nicolino
Just a reminder--there is still time to pre-register (deadline Friday, May 4th) for the 18th Annual Woods Hole Symposium, May 11-12th at Woods Hole Oceanographic Institute, Woods Hole, MA. This meeting is sponsored by the New England Society for Microscopy (NESM).
This meeting not only includes excellent biological and materials science speakers, but a symposium on "Remote Access Microsocpy" on Saturday morning.
I would like to direct listserver members to NESM's website which can be accessed directly (see below) or through the MSA website under affiliate societies. The url is: http://www.msa.microscopy.com/MSALAS/NESM/
Complete information re: the above mentioned symposium,including the program, plus registration forms and contacts can be found on this website under April Newsletter.
Hope to see many of you there!
Peggy Sherwood Corresponding Secretary, NESM -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
Hello- I am planning a periodic acid - Shiff stain for basement membrane in mouse skin. I understand there are two methods - one with alcoholic solutions and one with aqueous solutions. Does anyone have experience with this stain, and could you advise me in the choice of method? Any other advice would be quite welcome as well. Thanks!
Lara Muffley Dermatology Dept University of Washington Seattle, WA muffley-at-u.washington.edu
sample holders for JEOL 100CX-2000EXII model TEMs for sale:
gatan hot stage sample holders. both furnaces in working order; holders include one hexnut lockring AND thermocouple controller (sorry, no power wires--lost!): --#1 in good condition, $9000 OBO (gatan power supply model #580-0300) --#2 lightly stripped hex nut assembly (doesn't tighten all the way) in furnace, mA display on power supply (gatan power supply model #628-0500) a bit jumpy but TC reads temperature fine, $4000 OBO
extra washers (gatan part #628-0223) and hex nut tool (gatan part #608-0005) NOT included with the above holders.
Dera Postmaster You sent the wrong mail to wrong person. Pleases do not mail to me. My mail box are full. Kindly delete my name out of your groups mail. Thanks postmaster Best Reagrds, SU
Dr. Neuhaus, who contacted MSA and offered to donate a vintage light microscope, informs us that the instrument is on its way to a new home. See below.
I sense that he was a tiny bit overwhelmed by the size of the positive response he received.
Thanks, listers!!
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg ---------------------- Forwarded by Ronald Anderson/Fishkill/IBM on 05/02/2001 10:49 AM ---------------------------
"g. neuhaus" {ulmithaca-at-home.com} on 04/29/2001 06:47:28 PM
To: Ronald Anderson/Fishkill/IBM-at-IBMUS cc:
Below is the result of your feedback form. It was submitted by (dinesh-at-astra.iisc.ernet.in) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 3, 2001 at 05:19:42 ---------------------------------------------------------------------------
Email: dinesh-at-astra.iisc.ernet.in Name: Dinesh
Organization: Indian Institute of Science
Education: Graduate College
Location: Banagalore, Karnataka, India
Question: What is the wavelength and excitation required for identification of methane bacteria using fluorescence microscope.
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} Anyone done a tech time and cost analysis lately. Looking to compare notes. {BR} {BR} John Hoffpauir {BR} Cooper hospital {/FONT} {/HTML}
__________________________________________________ David Chiluiza USM, Institute of Marine Sciences Gulf Coast Research Laboratory 703 East Beach Drive, Ocean Springs, MS 39566-7000
Has anyone had experience with mounting an electron backscatter diffraction camera on an Amray FESEM 1845 round chamber? There is an EDS spectrometer already mounted on the chamber opposite the stage on the tilt side. This appears to leave a small port if the apertures can be relocated or a larger one if a fiber optic is feasible.
A friend of mine who makes excellent educational videos asked me a question today that I couldn't answer offhand. Will some old-timer please help my fading memory?
} ...when were the first EMs of chloroplasts (showing grana) produced? My } vague } guess was around 1949, but I can't find a definitive reference.
Please copy your response directly to him: bruce russell {biomedia-at-mail.telis.net}
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I am not at all familiar with the 1845. Way back when, we put BSE on our old JEOL U3. There were four solid state detectors that were fastened to the bottom of the pole piece with double stick tape. We also put BSE on our JEOL 840A. That was JEOL's system and it also fit to the bottom of the pole piece. Now we have a Robinson on a Hitachi 2460N which comes in from the side. We also have Oxford's Tetra with an array of detectors. Ours is set to slide in when needed.
I don't know why you couldn't mount a detector permanently to the pole piece and wire it to a feed thru on any port. Hopefully the BSE detector folks could work with you on this.
At 01:17 PM 5/3/2001 -0400, you wrote:
} Fellow listers, } } Has anyone had experience with mounting an electron backscatter diffraction } camera on an Amray FESEM 1845 round chamber? There is an EDS spectrometer } already mounted on the chamber opposite the stage on the tilt side. This } appears to leave a small port if the apertures can be relocated or a larger } one if a fiber optic is feasible. } } Thanks, } } Dave Audette } } david.audette-at-sylvania.com
Below is the result of your feedback form. It was submitted by (yarrabee-at-caboolture.net.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 3, 2001 at 19:52:32 ---------------------------------------------------------------------------
Email: yarrabee-at-caboolture.net.au Name: Eliza
Organization: QUT
Education: Undergraduate College
Location: Brisbane, QLD, Australia
Question: Hello. I was wondering if someone could tell me what the best method is of isolating mitchondria from rat liver cells for TEM analysis?
Below is the result of your feedback form. It was submitted by (barkdoll-at-acadia.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 3, 2001 at 23:43:49 ---------------------------------------------------------------------------
Question: I hope these questions are appropriate for this forum.
I am trying to planning to take high quality photomicrographs using a steromicroscope. I currently use a Russian made microscope and SLR body connected to an eyepiece tube. I have not been happy with this setup because of a noticeable degradation of image quaility towards the edges and difficulty of use. I am looking into upgrading to a system with fewer of these problems.
I have looked at literature from Nikon (SMZ series) and Olympus (SZ, SZH series) and received some quotes from distributors, however I am not tied to these manufacturers.
My questions are: 1) is there any source for "reviews" or published tests of particular microscopes? 2) how marked a difference should I expect between PLAN and PLAN-APO objectives? 3) my SLR does not have mirror lock-up - I'm willing to get a 35 mm back if warranted - how significant is the lack of MLU with an SLR an image quality? 4) some of the scopes are not inexpensive - what are some good ways for me to avoid getting a something that really exceeds my needs in image quality?
You have asked how to isolate rat liver mitochondria for TEM analysis. I ask why do you want to isolate them? If your interest is just in the ultrastructrure of the mitochondria themselves, then the easiest thing to do is fix and process the whole tissue. The liver is absolutely loaded with mitochondria and you will have plenty of them to look at. If your interest is more specific, then please elaborate on your project. Good luck in your endeavors, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
The enucleated eyes must be punctured at the ora serrata with a razor blade and then fixed by immersion in a mixture of 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer at pH 7.3 for 4-5 h. After removal of the cornea and lens the eyes are then rinse overnight in 0.1 M sodium cacodylate buffer containing 3% sucrose and post fixed in 2% osmium tetroxide in the same buffer for 2h at room temperature in the same buffer before dehydration and so on.
Ref: W.C. Yang, M.L. Hollenberg, and J.P.H. Wyse. Morphology of the retinal pigment epithelium in the vitamin A deficient rat. Virchows Arch. B Cell Path. 27, 7-21(1978)
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } "Quinn, Tim Lee" {tquinn-at-ku.edu} 04/30 10:22 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Fellow microscopists:
Does anyone have a good protocol for retina fixation for TEM?
I'm dealing with specimens collected in the field and it appears that the initial field fix- "cacodylate and glut" did not preserve the "rods and cones".
A good recomendation for a field fix would be appreciated. Would Karnovsky's work?
I also need to fix frog skin tissue, causing minimal shrinkage. Any suggestions?
I just recently read a job announcement, and it included that an EMSA certification would be preferred. Through some inquiries, I found that EMSA is the Electron Microscopy Society of America and I obtained a phone number for them (In Mass). However, I've searched the web and many microsopy sites and there seems to be no mention of this organization anywhere. This, along with the phone number being a private residence, leads me to believe this society has either renamed themselves or has disbanded. I was wondering if anyone has any information on ESMA or any other organization that offers a professional certification program for electron microscopy. If there are programs, I would also appreciate a critique on which are the best, and if a certification is even necessary for careers in electron microscopy. Any and all info is very helpful. Thanks.
Steve Wintonick
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com
Hi Guys, Can anyone give a list of activities that constitute routine maintenance of an SEM particulaly a LEO 1450 model with only secondary electron detector. Thanks Soji
Mr. O. O. ILORI DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING OBAFEMI AWOLOWO UNIVERSITY, ILE-IFE, OSUN STATE NIGERIA.
You may isolate mitochondria from homogenated tissue by using Percoll gradient. It's actually one step procedure. Percoll is iso-osmotic, it'll prevent mitochondria from damage. I don't remember details, but you have to mix your homogenate with buffer and Percoll in some proportion and centrifuge at low speed for about an hour. Than you will see the reddish mitochondria layer. You may remove Percoll by centrifugation at higher speed if necessary. I did some neg-staining of mitochondria couple of years ago. If you need detailed protocol, let me know, I have to check my old records.
Sergey.
} Date: Fri, 04 May 2001 10:04:25 -0400 } From: Timothy Schneider {Timothy.Schneider-at-mail.tju.edu} } Subject: RATS R US } To: microscopy-at-sparc5.microscopy.com } Reply-to: Timothy.Schneider-at-mail.tju.edu } X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2910.0) } Importance: Normal } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
If you are working with magnification of less the 80x you will probably have better luck with an extention bellows and a reversed Kodak Ektar Cine lens.
It will be a great deal easier on you pocket book as well.
Our eyes ignore a great deal of distortion that a camera quickly catches.
If you decide to buy a new scope or one new to you either test it at the dealers or buy it with the understanding that you can return it if it doesn't perform as you expect.
Stereo microscopes are not very good platforms for photography. Conventional macro photography methods are usually much better though less convineint.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} } Question: I hope these questions are appropriate for this } forum. } } I am trying to planning to take high quality photomicrographs using a } steromicroscope. I currently use a Russian made microscope and SLR } body connected to an eyepiece tube. I have not been happy with this } setup because of a noticeable degradation of image quaility towards } the edges and difficulty of use. I am looking into upgrading to a } system with fewer of these problems. } } I have looked at literature from Nikon (SMZ series) and Olympus (SZ, } SZH series) and received some quotes from distributors, however I am } not tied to these manufacturers. } } My questions are: } 1) is there any source for "reviews" or published tests of particular } microscopes? } 2) how marked a difference should I expect between PLAN and PLAN-APO } objectives? } 3) my SLR does not have mirror lock-up - I'm willing to get a 35 mm } back if warranted - how significant is the lack of MLU with an SLR an } image quality? } 4) some of the scopes are not inexpensive - what are some good ways } for me to avoid getting a something that really exceeds my needs in } image quality? } } Thank you very much, } Edwin Barkdoll V.M.D, Ph.D. } } ------------------------------------------------------------------------ --- }
Congratulations! You've already found "EMSA" even if you didn't realize it. EMSA became MSA, the Microscopy Society of America, a few years ago (1993? My, how time has flown!). You're already on the MSA listserver. The society homepage is at:
http://www.msa.microscopy.com/
For information on MSA certification, you can follow the links from MSA's homepage or from:
Certification is NOT necessary for a career in electron microscopy, but it may be required for a specific job opening. It does offer prospective employers some confirmation that you have the necessary background knowledge and skills to safely and successfully play with their expensive toys. The MSA certification is directed more toward biological microscopy (specifically biological TEM) than materials science microscopy.
Participation in some of the microscopy courses and training opportunities mentioned/promoted on the Listserver could also serve as assurance to prospective employers. I believe there are at least two colleges that offer degrees or certifications specifically in electron microscopy (one in California & one in Wisconsin???). Then there are places such as McCrone Research Institute that offer specialty microscopy degrees (e.g. Chemical Microscopy Certification at http://www.mcri.org/) which may include electron microscopy components along with light microscopy techniques.
There are also a few jobs like 'SEM technician - high school diploma required, SEM experience preferred' - a current, shiftwork job posting here in Massachusetts. Somehow I doubt that 'microscopist' will have any upward mobility unless he is VERY lucky and/or improves his education level.
Hope this has been helpful. I'm sure you'll get lots of other replies.
- Louise Harner
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-339-7300 phone 508-339-4996 fax Louise_Harner-at-albint.com
"steven wintonick" To: Microscopy-at-sparc5.microscopy.com {crimsem-at-hotm cc: ail.com} Subject: EMSA Certifcation???
2001/05/04 10:48 AM
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Hello,
I just recently read a job announcement, and it included that an EMSA certification would be preferred. Through some inquiries, I found that EMSA is the Electron Microscopy Society of America and I obtained a phone number for them (In Mass). However, I've searched the web and many microsopy sites and there seems to be no mention of this organization anywhere. This, along with the phone number being a private residence, leads me to believe this society has either renamed themselves or has disbanded. I was wondering if anyone has any information on ESMA or any other organization that offers a professional certification program for electron microscopy. If there are programs, I would also appreciate a critique on which are the best, and if a certification is even necessary for careers in electron microscopy. Any and all info is very helpful. Thanks.
Steve Wintonick
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com
Does anyone have any procedures to do Fine Needle Aspiration (FNA) for routine E.M. morphology. I.e.... collect, fix, rinse, dehydrate, infiltrate, and embed in an epoxy resin.
Ron Austin Dept of Pathology LSUMC Shreveport, LA rla-at-mindspring.com
Routine maintenance of an instrument like the LEO falls into the following areas -
1. Cleaning the cathode assembly each time you change a filament
2. Cleaning the anode every other time you change a filament
4. Using a chamois leather (washed oil free) to wipe round the gun chamber each time you change a filament
5. Keeping spare apertures for each part of the system (ask the service engineer or check in the manual).
5. Using a vacuum cleaner to clean up inside the apecimen area every few weeks.
That should keep you going but if you need more information our interactive Cd "Monitoring & Maintaining EM Performance" will tell you all you need to know about how the instrument works, which areas to check, how to check them and how to clean them if they are found to be a problem.
Good luck
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I have not used the Nikon stereo but I have used the Olympus SZ series and currently use a SZH10. The SZ has integrated objectives but will accommodate auxillary objectives. This scope is a good unit and is mostly a plastic-type unit. The SZH10 is metal and uses a screw-in objective or two objectives on a rotating turret. For most work, I really can't see any remarkable difference between Plan and PlanAPO objectives. I use a 1.0X PlanAPO on my SZH10 and 0.5X and 0.75X Plan objectives. The SZ scopes are rather good and are not horribly expensive. They do a good job.
I agree that trying take pix with an SLR is not only difficult but just about impossible--so to speak. The SLR must have a plain ground glass focusing screen or focusing will be a real challenge. It is a challenge even with the correct focusing screen. Mirror lock up and manual exposure is also highly desirable. The other problem (very subtle and quite frustrating) is that to get good pix with a stereo zoom, the metering system should be a spot meter type. The central portion of interest (assuming a non-uniform subject) will not properly expose (usually is way over exposed) when metered using normal averaging mode. So, spot meter, AE lock or manual mode, then mirror lock up and shoot.
The unevenness you may see on an SLR pix is due to vignetting as a result of the adapter between camera and scope. A different adapter should correct this.
If you are looking for truly high quality pix and want to do this with ease, I found that a camera system optimized for microphotography is necessary. For the Olympus line, the PM10-ADS is an automatic camera system with a leaf shutter. With this system, there is no shake at all. Unfortunately, I believe that this system has been discontinued. Their PM10-AD is an averaging meter system and is also discontinued. The PM-20 and PM-30 are newer systems and are quite expensive ($12K-$25K I think).
For digital cameras, I found that the Pixera units are outstanding. I use the Penguin 600CL. It is a cooled 5.8M pixel (2776x2074) camera which takes 17MB RGB TIFF and other format images in perfect rendition. This camera costs about $8500. The un-cooled version costs about $6600. The 1.5M pixel model costs about $6600. Their 1.2M pixel un-cooled camera (150ES) costs about $3700. These are complete camera systems which include a PCI card, real-time focusing interface, image capture and image viewing software for PC or Mac.
If you would like more information about any of these units, please contact me off-line.
Gary Gaugler, Ph.D. Optical Reflections 7970 Twin Rocks Rd Granite Bay, CA 95746-8111 916.791.8191 916.791.8186 fax
Disclaimer: I am an authorized dealer for these and other photographic and software products.
http://photoweb.net
At 06:38 AM 5/4/2001, you wrote:
Below is the result of your feedback form. It was submitted by (barkdoll-at-acadia.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 3, 2001 at 23:43:49 ---------------------------------------------------------------------------
Question: I hope these questions are appropriate for this forum.
I am trying to planning to take high quality photomicrographs using a steromicroscope. I currently use a Russian made microscope and SLR body connected to an eyepiece tube. I have not been happy with this setup because of a noticeable degradation of image quaility towards the edges and difficulty of use. I am looking into upgrading to a system with fewer of these problems.
I have looked at literature from Nikon (SMZ series) and Olympus (SZ, SZH series) and received some quotes from distributors, however I am not tied to these manufacturers.
My questions are: 1) is there any source for "reviews" or published tests of particular microscopes? 2) how marked a difference should I expect between PLAN and PLAN-APO objectives? 3) my SLR does not have mirror lock-up - I'm willing to get a 35 mm back if warranted - how significant is the lack of MLU with an SLR an image quality? 4) some of the scopes are not inexpensive - what are some good ways for me to avoid getting a something that really exceeds my needs in image quality?
The Microscopy and Microanalysis -2001 Program Search Engine is now on-line.
Using this search engine you can determine the time, location of and title of any presentation during the August Meeting in Long Beach.
You may reach this from the MSA WWW Site
http://www.msa.microscopy.com
then follow the links to the Microscopy & Microanalysis 2001 Meeting. You can also use it to view a detailed program listing for any day of the week, prior to receiving your hard copy of the program.
Regards to everyone. I am a graduate student at University of Southern Mississippi and I am seeking for a summer course in Scanning electron microscopy, preferably focused in biological analysis (I wook with fish larvae). If someone have knowledge of any course, please, let me know, I will thanks any information. Sincerely, David Chiluiza
__________________________________________________ David Chiluiza USM, Institute of Marine Sciences Gulf Coast Research Laboratory 703 East Beach Drive, Ocean Springs, MS 39566-7000
I think some of you will have an interest in the following, very sad news:
Prof. Dr. Ludwig Reimer (Muenster, Germany) died on April 29th, 2001.
I don't know how to express this correctly in english, but I feel it as a very big loss. I will leave it to others to review his scientific contributions to the community, I know him best as teacher to his students to whom he was always devoted.
Just wanted to let you know...
Petra Wahlbring -------------------------------------
Dr. Petra Wahlbring Centre de Recherche Gabriel Lippmann Laboratoire d'Analyse des Matériaux 162a, avenue de la Faiencerie L-1511 Luxembourg petra.wahlbring-at-crpgl.lu
I need to be away for two weeks, and would like opinions on what is best to do with an EDS detector, LN2-wise, while away. My options appear to be:
1. Leave everything on as normal. Don't really want to do this - I have someone I trust to keep the LN2 dewar full, but not really handle the microscope if something like a power failure or worse occurs.
2. Leave scope off, keep LN2 in EDS, hope residual vacuum in SEM is adequate for two weeks. I'm guessing it's best to power off the detector and computer, regardless.
3. Power down EDS, running detector through it's conditioning cycle, leave everything off and at room temperature.
4. Stay home, chained to the system.
Thoughts, experiences greatly appreciated.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
It's truly a big loss to electron microscopy community. Last mid-night, I just bought the new addition of his famous book "Transmission Electron Microscopy : Physics of Image Formation and Microanalysis (4th Ed)(Springer Series in Optical Sciences, Vol 36)" from Amazon.com. Didn't realize that it would be his last addition of the book. -cy
-----Original Message----- } From: Petra Wahlbring [mailto:petra.wahlbring-at-nexgo.de] Sent: Monday, May 07, 2001 2:03 AM To: Microscopy-at-sparc5.microscopy.com
Dear Colleagues,
I think some of you will have an interest in the following, very sad news:
Prof. Dr. Ludwig Reimer (Muenster, Germany) died on April 29th, 2001.
I don't know how to express this correctly in english, but I feel it as a very big loss. I will leave it to others to review his scientific contributions to the community, I know him best as teacher to his students to whom he was always devoted.
Just wanted to let you know...
Petra Wahlbring -------------------------------------
Dr. Petra Wahlbring Centre de Recherche Gabriel Lippmann Laboratoire d'Analyse des Matériaux 162a, avenue de la Faiencerie L-1511 Luxembourg petra.wahlbring-at-crpgl.lu
In your situation, and assuming I absolutely trusted the assistant with keeping the Liq. N2 up, I would use your option 2. Certainly turn off the detector and computer.
Tony Garratt-Reed
} Hi Listers, } } I need to be away for two weeks, and would like opinions } on what is best to do with an EDS detector, LN2-wise, while } away. My options appear to be: } } 1. Leave everything on as normal. Don't really want to do this - I have } someone I trust to keep the LN2 dewar full, but not really handle the } microscope if something like a power failure or worse occurs. } } 2. Leave scope off, keep LN2 in EDS, hope residual vacuum in SEM } is adequate for two weeks. I'm guessing it's best to power off the } detector and computer, regardless. } } 3. Power down EDS, running detector through it's conditioning cycle, } leave everything off and at room temperature. } } 4. Stay home, chained to the system. } } Thoughts, experiences greatly appreciated. } } Cheers, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman }
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
I think some of you will have an interest in the following, very sad news:
Prof. Dr. Ludwig Reimer (Muenster, Germany) died on April 29th, 2001.
I don't know how to express this correctly in english, but I feel it as a very big loss. I will leave it to others to review his scientific contributions to the community, I know him best as teacher to his students to whom he was always devoted.
Just wanted to let you know...
That is, indeed, very sad news. I met Ludwig when I was visiting the CNRS to learn EELS. Not only was he very helpful professionally, but we also shared many meals and as trip to Toulouse Latreck's birthplace. Bill Tivol
Could anyone help me with a good fixation protocol for marine fish larvae (1-5mm) for SEM? I have been using glutaraldehyde + Osmium tetroxide and in bloc staining with Uranyl acetate for TEM but I understand that osmolarity is a very important factor in SEM.
Do you think I can analyze in SEM the samples fixed for TEM?
Thanks a lot for any help. David Chiluiza
__________________________________________________ David Chiluiza USM, Institute of Marine Sciences Gulf Coast Research Laboratory 703 East Beach Drive, Ocean Springs, MS 39566-7000
----- Original Message ----- } From: "James M. Ehrman" {jehrman-at-mta.ca} To: "Microscopy Listserv" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, May 07, 2001 5:15 AM
A faculty member brought me some 10 year old samples embedded in Glycol-Methacrylate. He would like some thick sections 1-2microns. What's the best way to section and handle these to get good flat sections? Also is there any solvent which would dissolve the blocks so that the samples could be reembedded in another resin? All advice appreciated, thanks
Could anyone help me with a good fixation protocol for marine fish larvae (1-5mm) for SEM? I have been using glutaraldehyde + Osmium tetroxide and in bloc staining with Uranyl acetate for TEM but I understand that osmolarity is a very important factor in SEM.
Do you think I can analyze in SEM the samples fixed for TEM?
Thanks a lot for any help. David Chiluiza
__________________________________________________ David Chiluiza USM, Institute of Marine Sciences Gulf Coast Research Laboratory 703 East Beach Drive, Ocean Springs, MS 39566-7000
Does anyone know a good method for finding platinum in a cell? I am treating with platinum drugs and was wondering if I could localize platinum binding sites via TEM EDS analysis. Any help is appreciated!
Thank you,
Dale
-- ##################### Dale J. Telgenhoff # Zoology Department # Michigan State U. # (517) 355-3326 # telgenh4-at-msu.edu # #####################
"Give a man a fish and he will eat for a day. Teach him how to fish and he will sit in a boat & drink beer all day."
My answer is 3. Turn it off and let it warm up. Answer 2 is strange because the dewar vacuum is independent of the SEM vacuum. This what the window is for. If a windowless detector then no problem with warm up. See below
This answer assumes that system is reasonable new. Why? Because what happens when the EDS detector warms up is not a detector problem, it is a vacuum problem. If the vacsorb material inside the dewar warms up and it is full it will release the adsorbed gases to the dewar vacuum. New detectors (less than three years old) without obvious vacuum problems or degraded resolution now days can be warmed up without problems. In fact now days EDS systems are shipped dry by the manufacturers. We used to ship them cold in the seventies but that day has past. When I worked for Kevex and before that at Tracor Xray we saw very few problems when dewars warmed up except some freak problems. Freak problem did include windows blowing outwards and failure to cool down again. Then again I have met occasional EDS users who only cool the detector when they need to use it! They do fine.
Ron Vane XEI Scientific
-----Original Message----- } From: James M. Ehrman {jehrman-at-mta.ca} To: Microscopy Listserv {Microscopy-at-sparc5.microscopy.com}
I am a graduate student of Portland State University. I will take part in the Microscopy & Microanalysis, 2001 from August 5 to August 9. I will reserve room from the Westin Long Beach and want to share it with another male candidate.Please let me know as soon as possible once you are intrested in it.
You can get the information about Westin Long Beach from http://www.pkghlrss.com/liveres/res.asp
Base rate $139 (Nightly rates may vary.)
Sincerely,
Lifeng
Lifeng Dong Physics Department Portland State University P.O.Box 751 Portland,OR,97207-0751 U.S.A Tel:503-725-8061 Fax:503-725-3888
{html} {head} {/head} {body} Earl, {br} He's trying to have a life! The correct answer lies behind door #2. Just keep the dewar cold and the electronics off. Your vacuum system status is optional, depending upon the system and how well it protects itself. {br} Ken Converse {br} owner {br} Quality Images {br} third party SEM service {br} Delta, PA {br} {br} Earl Weltmer wrote: {br} {blockquote type="cite" cite="mid:000701c0d73c$90dbafe0$428ccd3f-at-pacbell.net"} {pre wrap=""} ------------------------------------------------------------------------ {br} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {br} To Subscribe/Unsubscribe -- Send Email to {a class="moz-txt-link-abbreviated" href="mailto:ListServer-at-MSA.Microscopy.Com"} ListServer-at-MSA.Microscopy.Com {/a} {br} On-Line Help {a class="moz-txt-link-freetext" href="http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html"} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {/a} {br} -----------------------------------------------------------------------. {br} {br} {br} Stay home is probably your safest bet. {br} {br} Earl {br} {br} ----- Original Message ----- {br} {/pre} {blockquote type="cite"} {pre wrap=""} From: "James M. Ehrman" {a class="moz-txt-link-rfc2396E" href="mailto:jehrman-at-mta.ca"} <jehrman-at-mta.ca> {/a} {br} {/pre} {/blockquote} {pre wrap=""} {!----} To: "Microscopy Listserv" {a class="moz-txt-link-rfc2396E" href="mailto:Microscopy-at-sparc5.microscopy.com"} <Microscopy-at-sparc5.microscopy.com> {/a} {br} Sent: Monday, May 07, 2001 5:15 AM {br} Subject: what to do with unattended EDS on SEM {br} {br} {br} {/pre} {blockquote type="cite"} {pre wrap=""} ------------------------------------------------------------------------ {br} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {br} To Subscribe/Unsubscribe -- Send Email to {a class="moz-txt-link-abbreviated" href="mailto:ListServer-at-MSA.Microscopy.Com"} ListServer-at-MSA.Microscopy.Com {/a} {br} On-Line Help {a class="moz-txt-link-freetext" href="http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html"} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {/a} {br} -----------------------------------------------------------------------. {br} {br} {br} Hi Listers, {br} {br} I need to be away for two weeks, and would like opinions {br} on what is best to do with an EDS detector, LN2-wise, while {br} away. My options appear to be: {br} {br} 1. Leave everything on as normal. Don't really want to do this - I have {br} someone I trust to keep the LN2 dewar full, but not really handle the {br} microscope if something like a power failure or worse occurs. {br} {br} 2. Leave scope off, keep LN2 in EDS, hope residual vacuum in SEM {br} is adequate for two weeks. I'm guessing it's best to power off the {br} detector and computer, regardless. {br} {br} 3. Power down EDS, running detector through it's conditioning cycle, {br} leave everything off and at room temperature. {br} {br} 4. Stay home, chained to the system. {br} {br} Thoughts, experiences greatly appreciated. {br} {br} Cheers, {br} {br} Jim {br} {br} -- {br} {br} James M. Ehrman {br} Digital Microscopy Facility {br} Mount Allison University {br} Sackville, NB E4L 1G7 {br} CANADA {br} {br} phone: 506-364-2519 {br} fax: 506-364-2505 {br} email: {a class="moz-txt-link-abbreviated" href="mailto:jehrman-at-mta.ca"} jehrman-at-mta.ca {/a} {br} www: {a class="moz-txt-link-freetext" href="http://www.mta.ca/~jehrman"} http://www.mta.ca/~jehrman {/a} {br} {br} {br} {br} {/pre} {/blockquote} {/blockquote} {br} {/body} {/html}
Below is the result of your feedback form. It was submitted by (kfield-at-fast.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, May 7, 2001 at 21:49:02 ---------------------------------------------------------------------------
Email: kfield-at-fast.net Name: Alex Field
Organization: Keith Valley Middle School
Education: 6-8th Grade Middle School
Location: Horsham, PA USA
Question: I am looking for a substage light or illuminator for an old Nikon Stereo microscope I was given. I want to be able to look at water samples from ponds at the organisms in the ponds near my home. Can you tell me where I can buy one?
As one of his students I feel a professional as well as a personal loss. I remember him as a great teacher, a good advisor, and I also recall many a bottle of Champaign when one of his students passed a final exam.
He will be missed...
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Petra Wahlbring [mailto:petra.wahlbring-at-nexgo.de] Sent: Monday, May 07, 2001 12:03 AM To: Microscopy-at-sparc5.microscopy.com
Dear Colleagues,
I think some of you will have an interest in the following, very sad news:
Prof. Dr. Ludwig Reimer (Muenster, Germany) died on April 29th, 2001.
I don't know how to express this correctly in english, but I feel it as a very big loss. I will leave it to others to review his scientific contributions to the community, I know him best as teacher to his students to whom he was always devoted.
Just wanted to let you know...
Petra Wahlbring -------------------------------------
Dr. Petra Wahlbring Centre de Recherche Gabriel Lippmann Laboratoire d'Analyse des Matériaux 162a, avenue de la Faiencerie L-1511 Luxembourg petra.wahlbring-at-crpgl.lu
I am working with an investigator who has been doing perfusion/fixation of rat placentas for EM immunolocalization. He is dissatisfied with the morphologic preservation and believes the problem is related to the mechanics of the perfusion.
Does anyone out there have experience doing these perfusions or know of someone that is that we could contact to get some advice on how to optimize the fixation procedure?
Thanks in advance for any help that you can offer.
Sincerely,
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
could you explain the rationale behind your suggestion?
I thought, the detector has to be kept cold for 2 reasons:
a) to keep the preamplifiers and detector cold for better S/N ratio b) to keep the ions (Li) in the crystal from diffusing in the electric field
If, and only if, this is correct, there is no need to keep the detector cold if the electronics and high voltage power supply are turned off. In fact, if you keep everything cold, it could attract dirt over time (cryo-trap), degrading the performance.
I agree with you on the vacuum, though. If you can, keep the scope under vacuum. If not, you may have to pump for a while to get out moisture, or perhaps even bake it out.
I could be wrong -- has happened before.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: {mailto:info-at-soft-imaging.com} web: {http://www.soft-imaging.com/} =================================== -----Original Message----- From: Ken Converse [mailto:qualityimages-at-netrax.net] Sent: Tuesday, May 08, 2001 4:51 AM To: Earl Weltmer; MSA, listserver Subject: Re: what to do with unattended EDS on SEM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------. Earl, He's trying to have a life! The correct answer lies behind door #2. Just keep the dewar cold and the electronics off. Your vacuum system status is optional, depending upon the system and how well it protects itself. Ken Converse owner Quality Images third party SEM service Delta, PA
Earl Weltmer wrote:
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} On-Line Help {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html}
----- Original Message ----- From: "James M. Ehrman" {jehrman-at-mta.ca} {mailto:jehrman-at-mta.ca} To: "Microscopy Listserv" {Microscopy-at-sparc5.microscopy.com} {mailto:Microscopy-at-sparc5.microscopy.com} Sent: Monday, May 07, 2001 5:15 AM Subject: what to do with unattended EDS on SEM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} On-Line Help {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html}
I need to be away for two weeks, and would like opinions on what is best to do with an EDS detector, LN2-wise, while away. My options appear to be:
1. Leave everything on as normal. Don't really want to do this - I have someone I trust to keep the LN2 dewar full, but not really handle the microscope if something like a power f! ailure or worse occurs.
2. Leave scope off, keep LN2 in EDS, hope residual vacuum in SEM is adequate for two weeks. I'm guessing it's best to power off the detector and computer, regardless.
3. Power down EDS, running detector through it's conditioning cycle, leave everything off and at room temperature.
4. Stay home, chained to the system.
Thoughts, experiences greatly appreciated.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Dear Listserver, Glycol methyacrylate can be cut easily with glass knives at 1 or 2 microns. The sections are like very fine cellophane and wrinkles can be spread out by placing them on a fairly deep(3-4inches) water bath...no heat. Then, they can be picked up using a paint brush onto clean glass slides and dried and stained.
Barbara L. Plowman Univ. of the Pacific School of Dentistry 2155 Webster St. San Francisco, CA email: Bplowman-at-sf.uop.edu Ph: 415-929-6692
Sorry. The website in the last e-mail is not very exact.In fact,you can get the information about Westin Long Beach directly from
www.pkghlrss.com/events/mm2001/mm2001.html
I am a graduate student of Portland State University. I will take part in the Microscopy & Microanalysis, 2001 from August 5 to August 9. I will reserve a room from the Westin Long Beach and want to share it with another male candidate.Please let me know as soon as possible once you are intrested in it.
Base rate $139 (Nightly rates may vary.)
Thanks,
Lifeng
Lifeng Dong Physics Department Portland State University P.O.Box 751 Portland,OR,97207-0751 U.S.A Tel:503-725-8061 Fax:503-725-3888
we work on polymers containing a small fraction of carboxylic or sulfonic acids or the respective metal carboxylates and sulfonates. We currently try to map these different metal ions used to neutralize these acid groups by means of EFTEM or x-ray mapping. The metal ions are thought to cluster and form small "aggregates" within a pretty homogeneous polymer matrix. Recently, it was suggested to calculate the contrast between the polymer matrix (a C and O or C, S, and O containg material) and the metal ions in order to ensure that we are not wasting our time trying something impossible. Now: HOW do I calculate the contrast between a metal ion and a polymer matrix ? References and procedures are very welcome !
Thanks in Advance
Andreas
************************************************* Dr. Andreas Taubert Dept. of Materials Science and Engineering 3231 Walnut Street University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
{html} {head} {/head} {body} Mike, one of the other replies to this answered the question quite well. What happens when you let it warm up is the zeolites in the dewar (this is actually a sorption pump) release what they have captured. This may then condense on the SiLi crystal, degrading resolution and sensitivity. If enough stuff has been captured by the zeolites, due to a leaking system, you will end up with more than 1 atmosphere in the dewar and blow your window outwards. {br} {br} When Kevex came out with their "super dry" detector, keeping it cold was not necessary, but if you turned the ION pump off, your warranty was null and void. Clean is the key and if you have a dewar with zeolites, that means cold, except to warm it up only enough to remove accumulated ice and then immediately cool it again. The manufacturers ship warm because the dewar was just pumped out and leak-tested, but my understanding is that you should not have it warm for more than a month total. {br} {br} Ken Converse {br} owner {br} Quality Images {br} third party SEM service {br} Delta, PA {br} {br} Mike Bode wrote: {br} {blockquote type="cite" cite="mid:D1B635C6092DD411B0700020781025DE0F6128-at-lakewood.soft-imaging.com"} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} Ken, {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} could you explain the rationale behind your suggestion? {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} I thought, the detector has to be kept cold for 2 reasons: {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} a) to keep the preamplifiers and detector cold for better S/N ratio {/span} {/font} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} b) to keep the ions (Li) in the crystal from diffusing in the electric field {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} If, and only if, this is correct, there is no need to keep the detector cold if the electronics and high voltage power supply are turned off. In fact, if you keep everything cold, it could attract dirt over time (cryo-trap), degrading the performance. {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} I agree with you on the vacuum, though. If you can, keep the scope under vacuum. If not, you may have to pump for a while to get out moisture, or perhaps even bake it out. {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} I could be wrong -- has happened before. {/span} {/font} {/div} {div} {/div} {div} {font face="Arial" color="#0000ff" size="2"} {span class="056345816-08052001"} mike {/span} {/font} {/div} {div} {/div} {p} {font face="Arial" color="#000000" size="2"} Michael Bode, Ph.D. {/font} {br} {font face="Arial" color="#000000" size="2"} Soft Imaging System Corp. {/font} {br} {font face="Arial" color="#000000" size="2"} 1675 Carr St., #105N {/font} {br} {font face="Arial" color="#000000" size="2"} Lakewood, CO 80215 {/font} {br} {font face="Arial" color="#000000" size="2"} =================================== {/font} {br} {font face="Arial" color="#000000" size="2"} phone: (888) FIND SIS {/font} {br} {font face="Arial" color="#000000" size="2"} (303) 234-9270 {/font} {br} {font face="Arial" color="#000000" size="2"} fax: (303) 234-9271 {/font} {br} {font face="Arial" color="#000000" size="2"} email: {a target="_blank" href="mailto:info-at-soft-imaging.com"} mailto:info-at-soft-imaging.com {/a} {/font} {br} {font face="Arial" color="#000000" size="2"} web: {a target="_blank" href="http://www.soft-imaging.com/"} http://www.soft-imaging.com {/a} {/font} {br} {font face="Arial" color="#000000" size="2"} =================================== {/font} {/p} {blockquote} {div class="OutlookMessageHeader" dir="Ltr" align="Left"} {font face="Tahoma" size="2"} -----Original Message----- {br} {b} From: {/b} Ken Converse [ {a class="moz-txt-link-freetext" href="mailto:qualityimages-at-netrax.net"} mailto:qualityimages-at-netrax.net {/a} ] {br} {b} Sent: {/b} Tuesday, May 08, 2001 4:51 AM {br} {b} To: {/b} Earl Weltmer; MSA, listserver {br} {b} Subject: {/b} Re: what to do with unattended EDS on SEM {br} {br} {/font} {/div} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to {a class="moz-txt-link-abbreviated" href="mailto:ListServer-at-MSA.Microscopy.Com"} ListServer-at-MSA.Microscopy.Com {/a} On-Line Help {a class="moz-txt-link-freetext" href="http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html"} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {/a} -----------------------------------------------------------------------. Earl, {br} He's trying to have a life! The correct answer lies behind door #2. Just keep the dewar cold and the electronics off. Your vacuum system status is optional, depending upon the system and how well it protects itself. {br} Ken Converse {br} owner {br} Quality Images {br} third party SEM service {br} Delta, PA {br} {br} Earl Weltmer wrote: {br} {blockquote cite="mid:000701c0d73c$90dbafe0$428ccd3f-at-pacbell.net" type="cite"} {pre wrap=""} ------------------------------------------------------------------------ {br} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {br} To Subscribe/Unsubscribe -- Send Email to {a class="moz-txt-link-abbreviated" href="mailto:ListServer-at-MSA.Microscopy.Com"} ListServer-at-MSA.Microscopy.Com {/a} {br} On-Line Help {a class="moz-txt-link-freetext" href="http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html"} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {/a} {br} -----------------------------------------------------------------------. {br} {br} {br} Stay home is probably your safest bet. {br} {br} Earl {br} {br} ----- Original Message ----- {br} {/pre} {blockquote type="cite"} {pre wrap=""} From: "James M. Ehrman" {a class="moz-txt-link-rfc2396E" href="mailto:jehrman-at-mta.ca"} <jehrman-at-mta.ca> {/a} {br} {/pre} {/blockquote} {pre wrap=""} {!----} To: "Microscopy Listserv" {a class="moz-txt-link-rfc2396E" href="mailto:Microscopy-at-sparc5.microscopy.com"} <Microscopy-at-sparc5.microscopy.com> {/a} {br} Sent: Monday, May 07, 2001 5:15 AM {br} Subject: what to do with unattended EDS on SEM {br} {br} {br} {/pre} {blockquote type="cite"} {pre wrap=""} ------------------------------------------------------------------------ {br} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {br} To Subscribe/Unsubscribe -- Send Email to {a class="moz-txt-link-abbreviated" href="mailto:ListServer-at-MSA.Microscopy.Com"} ListServer-at-MSA.Microscopy.Com {/a} {br} On-Line Help {a class="moz-txt-link-freetext" href="http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html"} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {/a} {br} -----------------------------------------------------------------------. {br} {br} {br} Hi Listers, {br} {br} I need to be away for two weeks, and would like opinions {br} on what is best to do with an EDS detector, LN2-wise, while {br} away. My options appear to be: {br} {br} 1. Leave everything on as normal. Don't really want to do this - I have {br} someone I trust to keep the LN2 dewar full, but not really handle the {br} microscope if something like a power f! {br} ailure or worse occurs. {br} {br} 2. Leave scope off, keep LN2 in EDS, hope residual vacuum in SEM {br} is adequate for two weeks. I'm guessing it's best to power off the {br} detector and computer, regardless. {br} {br} 3. Power down EDS, running detector through it's conditioning cycle, {br} leave everything off and at room temperature. {br} {br} 4. Stay home, chained to the system. {br} {br} Thoughts, experiences greatly appreciated. {br} {br} Cheers, {br} {br} Jim {br} {br} -- {br} {br} James M. Ehrman {br} Digital Microscopy Facility {br} Mount Allison University {br} Sackville, NB E4L 1G7 {br} CANADA {br} {br} phone: 506-364-2519 {br} fax: 506-364-2505 {br} email: {a class="moz-txt-link-abbreviated" href="mailto:jehrman-at-mta.ca"} jehrman-at-mta.ca {/a} {br} www: {a class="moz-txt-link-freetext" href="http://www.mta.ca/~jehrman"} http://www.mta.ca/~jehrman {/a} {br} {br} {br} {br} {/pre} {/blockquote} {/blockquote} {br} {/blockquote} {/blockquote} {br} {/body} {/html}
What a great venue! Is this list wonderful, or what?
Thank you, all, for the replies on lenses. I have kind of wondered about the markings for some time, but now I needed to understand what we have on the scope, and in the drawer.
To sum it up: Epi: Illumination can be supplied from above, through the objective.
Plan: Objective has a very flat field of focus.
Neofluar: Fluorite components for better spherical and chromatic corrections.
44 23 80: Zeiss catalog number for the lens.
MDPlan: Identified as "Metallurgical" & "Darkfield", but there is no darkfield light path in the objective. The Olympus web site lists the MDPlan as a brightfield objective, but does not indicate what "MD" stands for.
Yes, it really is a 150x/0.95 lens. We also have a 160x/0.95 Leitz Wetzlar lens that I did not mention.
IR: Lenses are ground and coated to optimize for infrared.
Two fantastic web sites were identified which really do answer more questions than were ever answered on any episode of "Soap".
www.micro.magnet.fsu.edu/primer/anatomy/specifications.html or www.micro.magnet.fsu.edu/primer/anatomy/index.html
www.microscopyu.com/articles/optics/objectivespecs.html or www.microscopyu.com
No, I did not think it was mundane, either. I just wanted to give warning to those that seem to be on a higher plane than me. I have an insatiable curiosity that has provided for me through the last 17 years of integrated circuit failure analysis.
Below is the result of your feedback form. It was submitted by (jms-at-vetmed.wsu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 8, 2001 at 18:14:38 ---------------------------------------------------------------------------
Question: I am a neuroscience grad student at WSU, interested in motor neurons in the spinal cord. I use different fluorscent tracers and labeled antibodies to visualize cells and proteins of interest through the microscope (we use a Leitz Diaplan). I am looking for information on cameras that are sensitive enough to visualize the fluorescence and transmit the images to my computer for image analysis (by optimas). What camera makes and models do you recommend? I use wide band UV, narrow band blue, and narrow band green filters most often. Thanks very much for any help you can give me!
} } I agree with you on the vacuum, though. If you can, keep the scope } under vacuum. If not, you may have to pump for a while to get out } moisture, or perhaps even bake it out. }
Bake it out?
Really?
Does anyone really do this/has anyone ever actually done it?
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
-- Hi, I am looking for a stain to use on unfixed cells and organelles so that I can see them throughout processing for EM (especially for cryo-sectioning). I believe such a stain is called a vital stain. Any help on this matter will be greatly appreciated. Many thanks Ken Blight. Senior Scientific Officer Imperial Cancer Research Fund London England.
University of Bristol in co-operation with Glaxo SmithKline
Department of Oral and Dental Science, Biomaterials Group
The Biomaterials Science Group in the Department of Oral and Dental Science, University of Bristol in collaboration with Glaxo SmithKline have a vacancy for a Postdoctoral Research Assistant in the Biomaterials Science Group on the project Interaction Mechanisms of Polymers at Interfaces of Mineralised Tissues.
The research area involves the study of the physical and chemical properties and interaction mechanisms of different polymers at interfaces of mineralised tissues. You will have recently been awarded a PhD in an appropriate field and will ideally have experience in scanning probe microscopy (AFM) of biological materials and other analytical techniques and an interest in medical research. You will work in Dr. Jandts group and interact with scientists at Glaxo Smith Kline.
The University of Bristol is one of the leading research universities in the UK and provides an outstanding scientific training environment to enhance your qualification. Bristol is located in the attractive south-west of England. The group is involved in exciting, interdisciplinary projects and maintains appropriate state of the art instrumentation. There exist opportunities for additional interactions with clinical scientists and other centres at the university.
We are looking for a dynamic and exceptionally well-qualified postdoctoral researcher who can interact effectively in an international and interdisciplinary team. The appointment will be on a Research Assistant 1A scale with a salary range of # 16775 to # 20465. This is a full time appointment and initially for one year.
Applicants should include a short CV, stating research experience and interests, publication list and addresses of two referees. The review of applications will start 24 May 2001 and will continue until the post has been filled.
For further details telephone ++44 117 954 6947, minicom ++ 44 117 928 8894 or E-Mail Recruitment-at-bris.ac.uk (stating postal address ONLY) quoting reference 7401. Applicants wishing to apply electronically must complete an "APPLICATION FORM FOR AN ACADEMIC VACANCY", found at http://www.bris.ac.uk/Depts/Personnel/recruit.htm
Closing Date May 24th, 2001
----------------------------------------------------------------- Dr. rer. nat. Klaus D. Jandt Senior Lecturer in Dental Materials Science and Biomaterials University of Bristol, Department of Oral and Dental Science Lower Maudlin Street, Bristol, BS1 2LY, UK Phone: +44-117-9284418, Fax: ++44-117-9284780 Internet: K.Jandt-at-bris.ac.uk WWW: http://www.dent.bris.ac.uk/Biomaterials/kdj.htm "We make Biomaterials Science work!" Imua a lanakila
I have 2 boxes of 10 each retipped tungsten filaments for an Amray 1200 up for grabs for the cost of shipping (our old SEM was destroyed in a plant explosion -- well, actually, a pressure vessel failure, but same end result.) Also included are column liner cleaning tools and a few bulbs. Please contact me directly if you're interested.
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
Is RMC still around? I have a RMC MT-7000 ultramicrotome that needs a tune up or something (it chatters during the cutting stroke). Does anybody have the number for whoever is taking care of RMC products now. Ventana?
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Colleagues, does anyone know of a similar film (Fuji ?) for Kodak Technical Pan Film 120? Ours has been on backorder close to a month and now we are being informed that not until tomorrow, maybe, may film arrive. Our TP120 is being used for the Zeiss EM 109. Many thanks for any responses Teresa
} Colleagues, does anyone know of a similar film (Fuji ?) for Kodak Technical } Pan Film 120? Ours has been on backorder close to a month and now we are } being informed that not until tomorrow, maybe, may film arrive. Our TP120 } is being used for the Zeiss EM 109. Many thanks for any responses Teresa
Rather than find another film/developer combination (with experimentation for correct exposure, aggravation, etc.), try another vendor for Tech Pan. Any professional camera store should have at least a dozen rolls in stock. B&H Photo and Video in New York City and Calumet in Chicago come to mind. They have 800 numbers for ordering and can ship overnight if you are in a real bind. Even if it is not in stock they can have Kodak ship a 'brick' (20 rolls I think) directly to you. Kodak TMax 100 is a fine-grain B&W film that would be, I think, the closest substititute. It does not have as much contrast as Tech Pan and I don't know how well it works with whatever developer you are using. Call Kodak (1-800-242-2424) and see what they recommend. Finding an alternate source for Tech Pan would be simpler than reconfiguring your exposure/film/development protocol.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
http://www.rmcproducts.com (520) 745-0001 Boeckeler Inst. Inc.
Good Luck, Sergey.
At 03:17 PM 5/9/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We - Electron Microscopy Sciences- have no problem to supply you this Kodak TP-120 film (#151-1054). Out catalog number for this is #74130 for 5 rolls lot and #74133 for 20 rolls lot. Phone: 800-523-5874. Web: www.emsdiasum.com
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} We - Electron Microscopy Sciences- have no problem to supply you this Kodak {BR} TP-120 film (#151-1054). Out catalog number for this is #74130 for 5 rolls {BR} lot and #74133 for 20 rolls lot. {BR} Phone: 800-523-5874. Web: www.emsdiasum.com {BR} {BR} Bang Nguyen {BR} EMS {/FONT} {/HTML}
Vickie,we have an excellent reference center for neuromuscular biopsies here at LSUHSC in New Orleans, LA (504) 599-0281 Cheryl Vega is the technologist. Teresa } Hello, } I am looking for a good reference lab that deals with muscle tissue. At } present we are using Athena Labs but are having horrendous tat. Do you use } any lab out there for } the more sophisticated tests done on frozen muscle? We do all the routine } tests in house. Please let me know what labs are avaliable. Thanks in advance } for your help. } Vickie Hackett
RMC is currently being supported by Boekeler Instruments. The guy to contact for service is Al Coritz, (800) 552-2262. He's a great guy and will set you up.
John Shields EM Lab University of Georgia jshields-at-cb.uga.edu On 9 May 2001, at 15:17, Paula Sicurello wrote:
} -------------------------------------------------------------------- } -- -- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- } -- -. } } } Hi Listers, } } Is RMC still around? I have a RMC MT-7000 ultramicrotome that needs } a tune up or something (it chatters during the cutting stroke). } Does anybody have the number for whoever is taking care of RMC } products now. Ventana? } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax }
To the person looking for work to be done on a RMC ultramicrotome: Contact Al Coritz at al-at-boeckeler.com Best of Luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
A vital stain is one that delineates living from dead cells. It penetrates dead ones and leaves living (unfixed) ones unstained. It may be that one of these (e.g., trypan blue) would work on your unfixed cells as some of them would be dead, but it would not be staining the cells of interest. You might be able to see them as a faint blue pellet because of the dead ones.
As for what stain would go inside living cells and not alter ultrastructure, I don't know. Sorry.
Sara Miller
Usually unstained cells look a little creamy, and you can see them in frozen preparations.
On Wed, 9 May 2001, ken blight wrote:
} Date: Wed, 9 May 2001 09:09:25 +0100 } From: ken blight {blight-at-icrf.icnet.uk} } To: microscorpy hints and tips {microscopy-at-sparc5.microscopy.com} } Subject: Vital stain } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } -- } Hi, } I am looking for a stain to use on unfixed cells and organelles } so that I can see them throughout processing for EM (especially for } cryo-sectioning). I believe such a stain is called a vital stain. } Any help on this matter will be greatly appreciated. } Many thanks } Ken Blight. } Senior Scientific Officer } Imperial Cancer Research Fund } London } England. } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I would like to thanks to all the people that have helped me with advises about larval fixation for SEM.
I have a question, a person told me that he has obtained good results fixing fish larvae with Karnovsky’s solution (a mixture of 8% paraformaldehyde, 0.2M sodium cacodylate buffer and 25% glutaraldehyde), but no one else has mentioned it. Please, let me know what do you think about this solution. I will have just one group of larvae this year and that will be all for my thesis (I cannot miss them). Last year I fixed larvae for TEM with glutaraldehyde + osmium tetroxide + uranyl acetate and I understand that this procedure obscure the sample surface and difficult observations.
Thanks again for your comments David Chiluiza
__________________________________________________ David Chiluiza USM, Institute of Marine Sciences Gulf Coast Research Laboratory 703 East Beach Drive, Ocean Springs, MS 39566-7000
Hi - I have an old Nikon Epiphot microscope that does not have a shield to protect the user from UV. Does anyone know of a spare parts company that would sell something this old? The other alternative is to have our shop fabricate one. We have been told that if we do that, it needs to be made of orange plexiglas. Does it need to be orange for protection? Thank you in advance for your help!
Lisa Wright Mercer University School of Medicine Division of Basic Medical Sciences, Research 1550 College Street Macon, GA 31207
I would be interested in the general comments from the microscopy community about how necessary such shields are. My Zeiss Axiophot didn't come with one, my Nikon Diaphot and Optiphot did and we use them, and my Olympus IX-70's did but we don't use it.
Not all plexiglas will stop UV equally well but clearly it doesn't have to be orange since they sell UV safety glasses for use DNA gels on UV light boxes that are clear plastic. Tom
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have had a Nikon Optiphot serviced annually for the past 12 years, and no one has ever mentioned the need for UV shielding. I'll be very interested to hear opinions about its necessity, too.
RMC is part of Boeckeler Instruments in Tucson Arizona. Last summer Ventana Medical Systems sold the product line to Boeckeler Instruments.Ventana's main focus was ISH & IHC in hospital histology labs and did not fit with their long term goals. We offer sales of new instruments as well as service on the old. Please feel free to contact me off line or visit ourwebsite listed below for further information.
Al Coritz Sales & Service Manager RMC Products Office: 520-745-0001 Fax: 520-745-0004 Cell: 520-465-3598 Al-at-boeckeler.com WWW.rmcproducts.com
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} Hi Paula & Everyone {BR} {BR} RMC is part of Boeckeler Instruments in Tucson Arizona. Last summer Ventana {BR} Medical Systems sold the product line to Boeckeler Instruments.Ventana's {BR} main focus was ISH & IHC in hospital histology labs and did not fit with {BR} their long term goals. We offer sales of new instruments as well as service {BR} on the old. Please feel free to contact me off line or visit ourwebsite {BR} listed below for further information. {BR} {BR} {BR} Al Coritz {BR} Sales & Service Manager {BR} RMC Products {BR} Office: 520-745-0001 {BR} Fax: 520-745-0004 {BR} Cell: 520-465-3598 {BR} Al-at-boeckeler.com {BR} WWW.rmcproducts.com {BR} {/FONT} {/HTML}
Dear Listers, I need your suggestions about the possiblity to buy a system based on the inverted microscope, but suitable for deconvolution and time-lapse analysis of fast events in living cells.
We would like to buy such system. Whinch one is the best (I mean quality devided on price)?
Sincerely yours, Alexander Mironov Consorzio Mario Negri Sud, Italy
Hi Vickie, Another excellent lab for muscle is located in Buffalo, New York. Their number is 719-878-7513. I would also be interested in the routine stains that you do in your lab and start a dialogue with you regarding the neuromuscular lab commonalities. F.Maiers
University of Bristol in co-operation with Glaxo SmithKline
Department of Oral and Dental Science, Biomaterials Group
The Biomaterials Science Group in the Department of Oral and Dental Science, University of Bristol in collaboration with Glaxo SmithKline have a vacancy for a Postdoctoral Research Assistant in the Biomaterials Science Group on the project Interaction Mechanisms of Polymers at Interfaces of Mineralised Tissues.
The research area involves the study of the physical and chemical properties and interaction mechanisms of different polymers at interfaces of mineralised tissues. You will have recently been awarded a PhD in an appropriate field and will ideally have experience in scanning probe microscopy (AFM) of biological materials and other analytical techniques and an interest in medical research. You will work in Dr. Jandts group and interact with scientists at Glaxo Smith Kline.
The University of Bristol is one of the leading research universities in the UK and provides an outstanding scientific training environment to enhance your qualification. Bristol is located in the attractive south-west of England. The group is involved in exciting, interdisciplinary projects and maintains appropriate state of the art instrumentation. There exist opportunities for additional interactions with clinical scientists and other centres at the university.
We are looking for a dynamic and exceptionally well-qualified postdoctoral researcher who can interact effectively in an international and interdisciplinary team. The appointment will be on a Research Assistant 1A scale with a salary range of # 16775 to # 20465. This is a full time appointment and initially for one year.
Applicants should include a short CV, stating research experience and interests, publication list and addresses of two referees. The review of applications will start 24 May 2001 and will continue until the post has been filled.
For further details telephone ++44 117 954 6947, minicom ++ 44 117 928 8894 or E-Mail Recruitment-at-bris.ac.uk (stating postal address ONLY) quoting reference 7401. Applicants wishing to apply electronically must complete an "APPLICATION FORM FOR AN ACADEMIC VACANCY", found at http://www.bris.ac.uk/Depts/Personnel/recruit.htm
2001 Closing Date May 24th, 2001
----------------------------------------------------------------- Dr. rer. nat. Klaus D. Jandt Senior Lecturer (Associate Professor) in Biomaterials Science University of Bristol, Department of Oral and Dental Science Lower Maudlin Street, Bristol, BS1 2LY, UK Phone: +44-117-9284418, Fax: ++44-117-9284780 Internet: K.Jandt-at-bris.ac.uk "We make Biomaterials Science work!"
Postdoctoral Position available: Biophotonics Instrumentation At University of Wisconsin-Madison, in the Instrumentation Laboratory of Dr. John White.
Postdoctoral Scientists are sought for the development of new optical instrumentation with an emphasis on spectral/lifetime imaging.
Previous experience in one or more of the following areas will be a primary consideration: optical design, non-linear optics, optical system development, techniques such as optical trapping or laser microsurgery. Experience with either confocal or multiphoton laser-scanning microscopy would be beneficial as the central emphasis of the study will involve these imaging techniques. Extensive facilities for microscopy, electrical engineering, software design and biological research are available for this project.
Ph.D.'s with multidisciplinary backgrounds from fields such as applied physics, biomedical engineering, electrical engineering, molecular/cell biology, nanotechnology or other related interdisciplinary fields are encouraged to apply.
Position is available as soon as June 1, 2001 with the application deadline open until the position is filled.
Please direct all questions via email to Kevin Eliceiri (eliceiri-at-facstaff.wisc.edu)
See www.loci.wisc.edu/employment.html for more information and application instructions.
-- Kevin W. Eliceiri
Lab Manager Microscopy Research Laboratory http://www.microscopy.wisc.edu
Project Director Laboratory for Optical and Computational Instrumentation (LOCI) http://www.loci.wisc.edu 271 Animal Sciences 1675 Observatory Dr. Madison, WI 53706 608-263-6288 voice 608-262-4570 fax
We currently have a 6 year old Sony video camera & printer with PAX-IT image capture software. We have had problems with the system over the years & poor support because the components are from different manufacturers (we bought it with an Olympus PLM & the computer was a hand me down from another lab section) so the manufacturers like to blame each other rather than fix the problem. We also found that the video camera gives very poor resolution above about 100X. We like the PAX-IT user interface when it works.
We are going to ask for a bridge comparison microscope with bright field, dark field, phase contrast, DIC, & simple pol. We also want to ask for a digital camera, new computer, & new or upgraded image capture software.
Because our analytical work always has the potential to go to court, I prefer software that does not allow the user to significantly alter the image (overlays, labeling, etc. are alright) like our current version of PAX-IT.
We will potentially use the camera & capture system on our other scopes (Wild M3 stereo, Olympus BX50P, AO 1-10 Compound, & Nikon Labophot Phase Contrast) & Polaroid MP-4 photo stand. (We currently use the video camera with a macro zoom lens on the MP-4.)
Our samples are mostly 'filth' (insect fragments, hairs, molds, yeast) in foods & some drugs.
Does anyone have any suggestion (positive or negative) about bridge comparison microscope models/brands, digital cameras, & image capture software?
David A. Foran, chemist Food and Drug Administration Kansas City District Lab DFORAN-at-ORA.FDA.GOV 913-752-2163 913-752-2727 (voice mail) 913-752-2151 (fax)
Dr. Halcrow has kindly modified/corrected my earlier statement on vital stains, but he was too polite to broadcast it to the net. I thought you should benefit from his comments and am sending them on at the risk of my embarassment. Anyway, my comments were based on what I was told many moons ago in grad school. In our usage, the stain colored dead or dying cells immediately, but was not taken up by viable cells, perhaps because we stained and viewed immediatley without allowing active uptake into the cells. From what I have read in the last few hours, it takes several minutes (30-60) for many vital stains to be taken up by living cells; also, cells (e.g., bacteria stained with janus green B) can go back to colorless in 24 hr.
Based on this exchange, I looked up viral stains and offer the following:
Stedmann. Vital s: applied to cells or parts of cells while they are still living.
Dorland. Vital s: staining of a tissue by a dye which is introduced into a living organism and which, by virtue of affinity for certain tissues, will stain those tissues (e.g., Conn: trypan blue injected IV stains kidney tubules)
Thompson. Bismark brown(s) Y, R, 53A, 53B, 53C; Janus green B; acridine orange; acridine red.
HJ Conn's Biological Stains by RD Lillie Selected Histochemical and Histopathological Methods by SW Thompson Dorland and Stedman are medical dictionaries.
---probably more than you ever wanted to know about vital stains.
Sara
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
---------- Forwarded message ----------
It is a bad idea to expose your skin to the UV produced by a mercury vapour lamp. But, assuming the lamp housing itself is light-tight, UV shields are only required if UV excitation is being used. If you are working with FITC (blue), GFP (blue) or RITC (green) type filters UV is not used for excitation, and the UV shield is unnecessary. Most pale yellow, yellow orange or red plastics absorb UV. Plexiglass in these colours should be readily obtainable - sign makers may have offcuts. If you cannot obtain yellow or orange plexiglass use an acetate filter of the type used for stage lighting. Chris
} ----- Original Message ----- } From: "Tom Phillips" {PhillipsT-at-missouri.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Thursday, May 10, 2001 8:32 PM } Subject: Re: LM shield for protection from UV } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } I would be interested in the general comments from the microscopy } } community about how necessary such shields are. My Zeiss Axiophot } } didn't come with one, my Nikon Diaphot and Optiphot did and we use } } them, and my Olympus IX-70's did but we don't use it. } } } } Not all plexiglas will stop UV equally well but clearly it doesn't } } have to be orange since they sell UV safety glasses for use DNA gels } } on UV light boxes that are clear plastic. Tom } } } } } } } } } } --------------------------------------------------------------------- } --- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } --------------------------------------------------------------------- } --. } } } } } } } } } Hi - } } } I have an old Nikon Epiphot microscope that does not have a shield } to } } } protect the user from UV. Does anyone know of a spare parts } company } } } that would sell something this old? The other alternative is to } have our } } } shop fabricate one. We have been told that if we do that, it needs } to be } } } made of orange plexiglas. Does it need to be orange for protection? } } } Thank you in advance for your help! } } } } } } Lisa Wright } } } Mercer University School of Medicine } } } Division of Basic Medical Sciences, Research } } } 1550 College Street } } } Macon, GA 31207 } } } } -- } } Thomas E. Phillips, Ph.D. } } Associate Professor of Biological Sciences } } Director, Molecular Cytology Core Facility } } } } 3 Tucker Hall } } Division of Biological Sciences } } University of Missouri } } Columbia, MO 65211-7400 } } (573)-882-4712 (voice) } } (573)-882-0123 (fax) } } }
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Does any one know about the concept of digital watermarking for proving the authenticity of an image?
David A. Foran, chemist Food and Drug Administration Kansas City District Lab DFORAN-at-ORA.FDA.GOV 913-752-2163 913-752-2727 (voice mail) 913-752-2151 (fax)
In general, real UV should not come though glass optics. In order to do so the optics should be made from quartz like quartz cuvettes for spectrophotometers. Unless the optic is not quartz there is no worry about UV. Some "near-UV" (300 and above nm) light could come through. Excitation wavelength for fluorescein is 490 nm (with emission at 520 nm) - far away from UV. Therefore I don't see any reason manufacturers will design microscope to pass real UV. Technically it's very difficult perform (and extremely expensive - you will easy know if your microscope is equipped with quartz optics).
Sergey.
At 02:32 PM 5/10/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
164 Ferne Court Palo Alto, CA 94306 (650) 494-0472 [NelsonC51-at-excite.com]
May 11, 2001
Hi David ... While I have never dealt with marine larvae of any kind (my M.A. thesis project concerned some morphological aspects of ciliates), I have dealt with Karnovsky's fixative. It's probably been suggested because the double effect of formaldehyde and glutaraldehyde preserve certain features (most likely, of the morphological type) better than either of those alone. However, that fixative also has a high osmolarity, and I'm guessing that this fixative may not work with marine animals. Good luck with your thesis project. Nelson Conti
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
They do modify slightly some particular pixels in your picture. This modification could not be recognizable by eye, but may be extracted using special algorithm (as Adobe Photoshop Plug-In do it). This "mark" will survive even after hard modification of the original picture, because it's not an "image" it's more like special "pattern" spreaded across the image. They claimed that even on very bad printed hardcopy that "water mark" will survive as well as on cropped/changed images. Is it not amusing! Watermark's "reader" is free, but you have to pay in order to be able to "mark" your image.
Sergey
At 06:27 PM 5/11/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } Dear David } } They do modify slightly some particular pixels in your picture. This } modification could not be recognizable by eye, but may be extracted using } special algorithm (as Adobe Photoshop Plug-In do it). This "mark" will } survive even after hard modification of the original picture, because it's } not an "image" it's more like special "pattern" spreaded across the } image. They claimed that even on very bad printed hardcopy that "water } mark" will survive as well as on cropped/changed images. Is it not } amusing! Watermark's "reader" is free, but you have to pay in order to be } able to "mark" your image. } } Dear David,
I wold approach the field of digital forensics with extreme caution. In the beginning I wold back up every image with a B&W shot of Color Slide methods that have a substantial body of work backing them up. A slick lawyer and painshop can morph you evidence into Porkies Hooter bar and grill sign so fast is leaves you in the dust. And you will like the warm unretouched glow that comes form film or slides. Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Although it might seem early to talk about the Microscopy and Microanalysis meeting in the year 2002 Quebec City, Canada, it is the time where the symposia are established and the symposia organizers determined.
In order to provide the best possible program, I, as program chair, am soliciting suggestions for symposia for the year 2002. Your idea for a great topic in any of the areas of "Physical Sciences," "Biological Sciences," or in "Advances in Instrumentation and Techniques," would be highly appreciated; after all, it is our annual meeting. Between now and June 30th 2001, I can be contacted under the following address:
mm2002-at-ornl.gov
Please forward the request also to your colleagues. With best regards,
Edgar Voelkl Program Chair M&M 2002
P.S.: For more information on the 2002 program, see: http://msc.rsvs.ulaval.ca/2002/2002.html
Below is the result of your feedback form. It was submitted by (bak-at-one.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, May 12, 2001 at 18:58:42 ---------------------------------------------------------------------------
Email: bak-at-one.net Name: Barbara Kostelnik
Organization: Hippo-Logistics
Education: Graduate College
Location: Cincinnati, OH USA
Question: I want to see if there are different visible pigment patterns in various colors of horse hairs as alleged by Dr. Ben Green in his controversial book, The Color of Horses. Here are some scanned images from his work: http://www.hippo-logistics.com/equus/Dr.Green/images/buckskin.jpg http://www.hippo-logistics.com/equus/Dr.Green/images/sorrel.jpg
One of our groups is using an old projecting microscope made by Bioscope Manufacturing. I just ordered an even older version on the internet to try myself. What equipment would YOU suggest to see if these patterns exist? (So far it looks to us like they don't.)
Thank you very much! Barbara Kostelnik Hippo-Logistics.com
I agree with Barbara that you can cut these blocks with as good a result as in new blocks, but in my experience (specifically with JB-4) the ease of cutting depends on the size of the block and how dry it is. Too dry a block will cause chatter and too wet a block will give you mushy looking sections. You can put the blocks in a chamber with either a container of water or desiccant to change this factor (don't touch either to the block). Also with JB-4, the hydrated sections can only be "drawn" out of the water on the slide, as they stick to anything they touch once their wet.
As for removing the material, I don't know of any successful method for this on JB-4 blocks. Maybe it is possible on other types of glycol methacrylate.
Karen Pawlowski, Ph.D. UT Dallas, Dallas, TX
Barbara Plowman wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listserver, } Glycol methyacrylate can be cut easily with glass knives at 1 or 2 microns. The sections are like very fine cellophane and wrinkles can be spread out by placing them on a fairly deep(3-4inches) water bath...no heat. Then, they can be picked up using a paint brush onto clean glass slides and dried and stained. } } Barbara L. Plowman } Univ. of the Pacific } School of Dentistry } 2155 Webster St. } San Francisco, CA } email: Bplowman-at-sf.uop.edu } Ph: 415-929-6692
I did not make the point I wanted in my last post about digital images in court. Put your self on the stand and answer these questions. Sir, did you modify the image in any way? Sir, is the image a true representation of the object. Sir, does each pixel have the same response to light as every other pixel? [three questions with mutually exclusive answers] Sir, how can you prove to me that this image was not manipulated before you watermarked it.
After you testimony the defenses expert explains the problem of having to apply a LUT table to the image to get at better image because different pixels have different gain and then gives a demo on image manipulation.
None of this comes up if you have a conventional silver print. If you need to use image enhancement to show something you can show it in relation to a negitive that can be examined by experts for modification. While digital image can be examined for modifcation explaing it to a jury will be very difficult to get them to understand after being shown how much an image can be changed.
The day will come that digital images are defensible in court but I would much rather be in the position of attacking one than defending one today. With all the bad press coming out about the FBI lab and other forensic labs I would be extremely careful to leave no opening for a crack in the chain of evidence.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
One thing that comes to mind is teaching new dogs old tricks.
As technology advances, the devices of 30, 50 and more years ago are forgotten for the most part, however, for reasons of economy preservation or nostalgia, some people like to keep, restore and maybe even use these antiquated devices. It might be helpful and interesting to at least collect information on the care and feeding of an old microscope, techniques of micrography with a non-automatic camera and the best methods to follow when trying to find and/or make substitutions for parts on aging equipment.
Ron L ----- Original Message ----- } From: "Dr. Edgar Voelkl" {mm2002-at-ornl.gov} To: {Microscopy-at-sparc5.microscopy.com} Cc: {3dem-at-sdsc.edu} Sent: Sunday, May 13, 2001 11:59 AM
The most prominent digital watermarking company is Digimarc http://www.digimarc.com
Digital watermarking is under the moniker "steganography." It is quite interesting and definitely a challenge to accomplish without disturbing the original (creating artifacts). Unfortunately, Digimarc's method is not very resilient. It can be easily removed using a relatively simple C program.
Advise that you tread lightly in this area until and unless new, more robust technology becomes available.
gary g.
At 03:27 PM 5/11/2001, you wrote:
} Does any one know about the concept of digital watermarking for proving the } authenticity of an image? } } David A. Foran, chemist } Food and Drug Administration } Kansas City District Lab } DFORAN-at-ORA.FDA.GOV } 913-752-2163 } 913-752-2727 (voice mail) } 913-752-2151 (fax)
I am not advocate for "watermarking". Digital "watermarks" developed for purpose to identify the "image creator" (who owned un-touched original file). And the idea is that "watermark" may survive on hardly modified image identified therefore the author. For instance, if somebody stole image file from your WEB site and use it to create some poster and sell it, you my prove that your image was used without authorization if "watermark" exist. It's not my personal opinion, it how I understand the purpose of "watermarking" from company who offered it (it's Adobe related, I believe). As I mention in my original message, company claims that it's possible to extract watermark from printed hard copy, poster in my example.
Nice negatives/slides are always a plus, but they are not protected from fraud and fire. Watermarks is not about image quality, it's about some security. You may keep your beautiful slides in achieve, but you must to convert it in digital if you want to post it on Internet. Here watermarks may work.
I don't see any reasons at all to use watermarks in forensics, but artists, publishers may be interested.
Sergey
At 02:44 AM 5/13/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Watermarks are great if you spend eons composing or editing a scanned image and some hoser rips it off, puts it on his site and claims it is his! the water mark allows you to quickly ID it.
Of course there are other ways to handle such scoundrels as well ... but that will be left for another dissertation!
Ed Sharpe archivist for SMECC
} Subj: Re: digital watermarking } Date: 5/13/01 6:48:14 PM US Mountain Standard Time } From: sryazant-at-ucla.edu (Sergey Ryazantsev) } To: gcouger-at-couger.com (Gordon Couger) } CC: microscopy-at-sparc5.microscopy.com } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Gordon } } I am not advocate for "watermarking". Digital "watermarks" developed for } purpose to identify the "image creator" (who owned un-touched original } file). And the idea is that "watermark" may survive on hardly modified } image identified therefore the author. For instance, if somebody stole } image file from your WEB site and use it to create some poster and sell it, } you my prove that your image was used without authorization if "watermark" } exist. It's not my personal opinion, it how I understand the purpose of } "watermarking" from company who offered it (it's Adobe related, I believe). } As I mention in my original message, company claims that it's possible to } extract watermark from printed hard copy, poster in my example. } } Nice negatives/slides are always a plus, but they are not protected from } fraud and fire. Watermarks is not about image quality, it's about some } security. You may keep your beautiful slides in achieve, but you must to } convert it in digital if you want to post it on Internet. Here watermarks } may work. } } I don't see any reasons at all to use watermarks in forensics, but artists, } publishers may be interested. } } Sergey } } At 02:44 AM 5/13/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } }
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} Watermarks are great if you spend eons composing or editing a scanned image {BR} and some hoser rips it off, puts it on his site and claims it is his! {BR} the water mark allows you to quickly ID it. {BR} {BR} Of course there are other ways to handle such scoundrels as well ... but that {BR} will be left for another dissertation! {BR} {BR} Ed Sharpe archivist for SMECC {BR} {BR} {BR} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} Subj: {B} Re: digital watermarking {/B} {BR} Date: 5/13/01 6:48:14 PM US Mountain Standard Time {BR} {I} From: sryazant-at-ucla.edu (Sergey Ryazantsev) {BR} To: gcouger-at-couger.com (Gordon Couger) {BR} CC: microscopy-at-sparc5.microscopy.com {BR} {/I} {BR} {BR} {BR} {BR} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} -----------------------------------------------------------------------. {BR} {BR} {BR} Dear Gordon {BR} {BR} I am not advocate for "watermarking". Digital "watermarks" developed for {BR} purpose to identify the "image creator" (who owned un-touched original {BR} file). And the idea is that "watermark" may survive on hardly modified {BR} image identified therefore the author. For instance, if somebody stole {BR} image file from your WEB site and use it to create some poster and sell it, {BR} you my prove that your image was used without authorization if "watermark" {BR} exist. It's not my personal opinion, it how I understand the purpose of {BR} "watermarking" from company who offered it (it's Adobe related, I believe). {BR} As I mention in my original message, company claims that it's possible to {BR} extract watermark from printed hard copy, poster in my example. {BR} {BR} Nice negatives/slides are always a plus, but they are not protected from {BR} fraud and fire. Watermarks is not about image quality, it's about some {BR} security. You may keep your beautiful slides in achieve, but you must to {BR} convert it in digital if you want to post it on Internet. Here watermarks {BR} may work. {BR} {BR} I don't see any reasons at all to use watermarks in forensics, but artists, {BR} publishers may be interested. {BR} {BR} Sergey {BR} {BR} At 02:44 AM 5/13/01 -0500, you wrote: {BR} >------------------------------------------------------------------------ {BR} >The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} >To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} >On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} >-----------------------------------------------------------------------. {BR} > {BR} > {BR} > {BR} {/BLOCKQUOTE} {BR} {BR} {/FONT} {/HTML}
The 17th Australian Conference on Electron Microscopy will be held in the Adelaide Convention Centre 4th - 8th February 2002, presenting you with an opportunity to escape cold climes and enjoy the worlds best fresh food and wines whilst participating in a conference with, the majority of Australia's and some of the World's leading Electron Microscopists, Scanned Probe Microscopists, and Confocal Microscopists.
The Australian Society for Electron Microscopy (Inc.) is the host society for this meeting in Adelaide, which is the is the Capital of the State of South Australia. Adelaide is a picturesque city and is the business and educational centre for the state. South Australia is noted for its glorious Mediterranean climate, picturesque surfing beaches, aquaculture, unpolluted agriculture and of course, 5 of the worlds truly great wine producing areas.
There are many unique and significant tourist destinations are within easy reach of Adelaide. The Barossa Valley with its unique German influence, Southern Vales and the Clare Valley are wine regions producing Australia's most famous wines such as the Grange and Hill of Grace. The rugged Flinders Ranges, Kangaroo Island, the Adelaide Hills and Beaches, offer a variety of scenic and relaxing outings to take at your convenience.
Planning for the meeting is well advanced and you will be receiving registration of interest and calls for abstracts in the near future. We have focused on the website as our major form of communication. I do hope that you can visit the site from time to time to maintain an awareness of the ACEM17 activities. (http://www.adelaide.edu.au/CEMMSA/acem17)
The conference will incorporate a series of workshops covering a broad range of topics. Student microscopists are encouraged to attend to learn or to launch their presentation careers and a series of student bursaries will be offered to facilitate this. On behalf of the Organising Committee may I extend an invitation to attend this important meeting.
-- John Terlet Director CEMMSA (Centre for Electron Microscopy & MicroStructure Analysis) Adelaide University Ph: 61 8 8303 5855 Fax: 61 8 8303 4356 http://www.adelaide.edu.au/CEMMSA
In an earlier post there was a mention of the images being used in forenic work. Then the watermark question. I sould have mentioned the forenic post in my reply about watermarks.
My point being that I would be very nervous using a digital image in court because of the ease that they can be manupulated and the fact that there is no way to tell if it is orignal or not.
The check needs to be made payable to Penn State University not PUS. Rosemary
} Rosemary, I talked to Stacie. She will send you a check for $2,400.00 next } week. The check will be made payable to PUS. Is all this OK? Thank you. } } Hong } ---------- } } From: Rosemary Walsh {rw9-at-psu.edu} } } To: Hong Yi {hyi-at-emory.edu} } } Subject: Fund transfer to cover dinners } } Date: Fri, May 11, 2001, 11:24 AM } } } } } Hi Hong, } } After consulting with our admin. assistant, it would be best } } to have Stacie send me a check for $1160 } } so that I can sign contracts and purchase sodas and supplies locally. } } Rosemary } } -- } } Rosemary Walsh, Manager } } The Electron Microscope Facility for the Life Sciences, } } A Shared Technology Facility, The Life Sciences Consortium } } 1 South FrearLab } } Penn State University } } University Park, PA 16802 } } (814) 865-0212 } } rw9-at-psu.edu } } http://www.lsc.psu.edu/stf/em/home.html
-- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
} That is fine with me, Hong. I have given Stacie the information on } where to send it. Rosemary
} Rosemary, I will have Stacie to cut you a check for $2,400.00. That will } include the fee for using your facility and food money. Is that OK with } you? Let me know so I can call Stacie soon. Thank you. } } Hong } ---------- } } From: Rosemary Walsh {rw9-at-psu.edu} } } To: Hong Yi {hyi-at-emory.edu} } } Subject: Fund transfer to cover dinners } } Date: Fri, May 11, 2001, 11:24 AM } } } } } Hi Hong, } } After consulting with our admin. assistant, it would be best } } to have Stacie send me a check for $1160 } } so that I can sign contracts and purchase sodas and supplies locally. } } Rosemary } } -- } } Rosemary Walsh, Manager } } The Electron Microscope Facility for the Life Sciences, } } A Shared Technology Facility, The Life Sciences Consortium } } 1 South FrearLab } } Penn State University } } University Park, PA 16802 } } (814) 865-0212 } } rw9-at-psu.edu } } http://www.lsc.psu.edu/stf/em/home.html
-- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
My son is doing a science project where he needs to contact a researcher working with Multiple Sclerosis and ask some questions. He has not had any responses on his previous email attempts to contact someone, so I'm turning to the listserver (even though it's a little off-topic).
Can anyone put me in touch with an MS researcher who would be able to take a few minutes to respond to questions from a 13 year old?
Thanks, Henk Colijn Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Can anyone account for the cause and solution to this problem?
Background:
We have installed a custom reticle in the eyepieces of our cytotechnologists' microscopes. Looking through the microscope, one sees a clear square area outlined by neutral density filter-like segments. The corners of the square just touch the boundaries of the field-of-view. In the center, there are a 6.7 micrometer diameter circle and 2 crossed 11 micrometer long lines for use as size comparators.
The square provides a user-friendly frame of reference by which a cytotechnologist can advance a Pap smear the length of 1 side while screening, using a 10x objective and 10x eyepieces. This systematic step-size movement promotes more complete and reproducible screening coverage, and initially has resulted in an increase in the pick-up rates for various categories of abnormal cells. The two size comparators are intended for use with a 40x objective to assist cytotechnologists when estimating nuclear areas and potential significance.
Problem:
One cytotechnologist in particular complains that her non-dominant eye "blurs over" when she encounters acellular fields-of-view. Despite patient persistence in working through this effect, she eventually gave up, as this effect was causing her to have headaches. I have removed the reticle. A few other cytotechnologists have complained similarly, but they have learned to tolerate the annoyance. The overwhelming majority of users have not experienced this problem.
Hypothesis:
I don't know why the reticle is causing this one user's eye to "blur over" (her words). However, I suspect that when both eyes are looking at cellular fields-of-view, both are focused at the same distance. However, upon encountering acellular fields, the dominant eye focuses on the two size comparators, while the non-dominant eye focuses at a different distance. Thus the brain is receiving mixed signals. To test this hypothesis, I've returned a reticle to the manufacturer to remove the size comparators. I'll reinstall this modified reticle and see whether the blurring over persists. If not, then problem solved. If yes, then the user will have to screen without the benefit of the reticle. I've also asked the lady to ask her ophthalmologist to write down her visual status.
Can anyone explain the cause and prevention of this phenomenon? Thanks.
The University of Missouri is searching for an Assistant or Associate Director of their Molecular Cytology Core Facility. This facility deals with all types of light microscopy. Individuals who feel they may be qualified for the position are encouraged to either apply or contact Tom Phillips for further details. A description of the job is appended to this message.
Assistant or Associate Director Molecular Cytology Core Facility
The Molecular Biology Program at the University of Missouri is seeking an individual with experience in some or all of the following areas:
* confocal scanning laser microscopy * bright field microscopy * wide field fluorescence microscopy * deconvolution * image processing/analysis * all types of microtomy * immunocytochemistry * in situ hybridization * Adobe Photoshop
The Assistant/Associate Director will be responsible for training users, maintaining instruments and developing protocols for a campus-wide multi-user facility. PhD desirable but not required for individuals with extensive experience. Although an ideal candidate would have experience in all of the areas listed above, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Electron microscopy is not a component of this core facility. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired.
The Core's web site can be viewed at http://www.biotech.missouri.edu/mbp/cores/index.html
Address applications (CV and 3 letters of reference) or inquires to:
Thomas E. Phillips, Ph.D. Division of Biological Sciences 3 Tucker Hall, University of Missouri Columbia, MO 65211-7400. 573-882-4712 PhillipsT-at-missouri.edu.
I am so sorry Gordon. I was answering the question of David Foran what digital watermark is. I did not figured out that the question was related to forensic. But, my position is still the same: "watermarks" are about copyrights protection and very slightly related to the image-quality. You may easily present to the court the image and nobody (except author) will know that it's watermarked. Using very simple program, author may verify his/her authority on this image at the court. Because this technology has been protected by at least 5 patents and registered, court would accept such evidence, I believe. This what purpose of digital watermarks in my opinion. The same way, some detective may watermark his images with some evidence and present it to court. Watermark in this case will not verify the trustness of the images, but authority of this particular person on the images. The same: some Laboratory (even forensic) may use watermarks to identify their digital products.
I do agree with you, that it's so easy to manipulate image digitally nowadays. From another hand, we do have an arsenal of the old nice tricks how to modify negatives/slides as well. People who prefer to stick to "film" will probably agree with me that it's very possible to modify images on the film too. Simplest example: using "ferro-cyanide" (not sure the spelling is correct) you may partially/completely eliminate part of the image on the wet negative/paper. After washing - the modification is undetectable in practice. Using this way, you could, for instance, remove some bands from electrophoresis pictures, or using multiple exposure - enhance some.
Sergey.
At 04:08 AM 5/14/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers. We are in the midst of trying to set up a new EM facility here. I am wondering how many of you out there actually support your facility with users fees? Is it realistic to expect service contracts, personnel salaries, supplies and maintenance can be recouped from fees? Or do most facilities need to be subsidized to ensure their survival? I worry that the kind of fees we would have to charge to ensure survival will scare many users away. Any suggestions, comments would be appreciated.
JoAnn Buchanan Dept. Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305
That's not as big an issue as a lot of people make it out to be. Yes, you have to show the provenance of an image in court, and be able to describe and defend the kind of processing you do.
However, the bottom line is that the image is *not* the testimony. The *testimony* is the testimony. It has *always* been possible to manipulate images; the ease in doing it is secondary. The important thing is that you have an expert sitting there and stating under oath that the image does or does not accurately represent what it is supposed to represent.
The assumption that the expert is lying unless the image itself can be independenly proven to accurately represent what the expert says is simply wrong. Images, except in specific circumstances, are there to *illustrate* observations made by the expert.
Consider scene photography. The homicide cop gets up and says there was a piece of paper on a table. There is a photograph of a piece of paper on a table. Counsel asks "Is this a fair and accurate representation of where the paper was?" The cop replies "Yes, it is."
Now, defense may get up and say "Aha! That's a digital image! Somebody could have *put* that image of paper on that table."
The answer is "Yeah, they could have. But the cop here says that's where it was. The cop here says that's where he saw it. The cop here says this here's what it looks like." The jury will either believe the cop or they won't.
One can think of cases where a photograph *doesn't* fairly and accurately represent what the cop saw -- the most common example I have seen is in color balance problems where the cop says the sweater was "green" and the photo shows it blue. Then you either don't enter it into evidence or you do the appropriate testimony. In other cases, image processing is integral to the conclusion itself -- such as edge enhancement or deblurring -- in which case you have to detail and defend the algorithms. But even then, that justification is based on testimony of the expert, not on some feature measurement of the image, and the "ease" with which it was done is fairly irrelevant.
If the court rejects testimony on the basis of some secondary measure of accuracy other than the testimony of the expert witness, then the lawyers have not done their job. The bullshit job that the defense did in the OJ trial is *not* the paradigm of how a trial should work, and should not be the standard for how evidence is examined.
billo
On Mon, 14 May 2001, Gordon Couger wrote:
} } } Dear Sergey } } } My point being that I would be very nervous using a digital image in court } because of the ease that they can be manupulated and the fact that there } is no way to tell if it is orignal or not. } } Gordon } } }
Below is the result of your feedback form. It was submitted by (rj.carey-at-student.qut.edu.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 15, 2001 at 02:44:36 ---------------------------------------------------------------------------
Email: rj.carey-at-student.qut.edu.au Name: richard carey
Organization: queensland university of technology
Education: Undergraduate College
Location: brisbane.queensland,australia
Question: A skin nodule resected from a patient is suspected of being a malignant melanoma.How would I prepare and view the sample to determine if this was the case?
I would appreciate from users of the Wacom PL series LCD writable tablet, for use specifically with image analysis, not graphics arts. The ability to trace directly on the image rather than using a mouse or pen/tablet combination seems appropriate but would like some feedback. TIA.
Walter F. Bobrowski Electron Microscopy Laboratories Drug Safety Evaluation Pfizer Global Research and Development 2800 Plymouth Road Ann Arbor MI 48105
Our SEM facility staggered along for years while supported partly from the grants of our five principal users and partly from user fees. This was not a stable system, since grant support dried up and usership was erratic.
Now our service contract and technician's salary are subsidized by three departments, and all members of these departments have free use of the facility and technical assistance. Fees from users outside of these departments are used to purchase supplies and for equipment repair/upgrades. These fees can be kept at a reasonable level, and members of the subsidizing departments love having free access to the lab. If you can arrange some sort of subsidy (and it is not too much of a burden if shared among departments), it is a great way to go.
Leslie Eibest
At 4:12 PM -0700 5/14/01, JoAnn Buchanan wrote: } Is it realistic to expect service contracts, personnel salaries, } supplies and maintenance can be recouped from fees? Or do most } facilities need to be subsidized to ensure their survival?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Joann, We will, hopefully, get started on a new pricing scheme in June. We have recently combined a small service facility from the Vet School with a multi-user facility that I have managed for, primarily, the School of Agriculture. The Ag. School has picked up all costs for service contracts and misc. to date including half of my salary (other half picked up by School of Science). Thus there was no charge, except for consumables, for use of this facility over the past 17 years. Now they want us to charge to try to recoop some of the costs. We run into problems that if you charge labor (for service) than you have to pay all the employees fringe benefits which are usually paid by the University. This adds about 1/3 to the cost of the employee to the department/school. Thus fees may actually cost the department/school more if you do not bring in enough money.
Thus there is the problem of how do you charge for service that will fairly compensate for the employee's time while still not penalizing the multi-user who works independently. Also, if you charge too much for imaging time you will tend to reduce use of equipment and it costs the same for service contracts whether booked full time or only part-time.
Consumables are not a problem since we can figure that fairly easily and multi-users provide most of their own reagents, etc.
I would like to consider an approach that would have departments pick up a share of the costs based on the number of faculty and their students who have used the facility over the last 6 months or so. They could pass that on to the researchers if they wanted to... Since we have over 15 departments who use this facility each year, an average of $1500 per department would go a long way to covering service contracts...especially if the Deans of the 5 schools involved would also toss in another couple of thousand each. Consumable reimbursement and very modest beam charges (~$10/hr) would cover misc. expenses while charge for labor for those using service would help defray costs of one technical person.
I would be very interested to hear if anyone has tried a plan such as this to help cover costs. I do know that it is used at one university in a large computer lab. Everyone who uses the lab is charge $50/month. Thus they have essentially unlimited use for that month. If they only use the facilities of 1 month, they are charged accordingly. If more than 1 person from a lab uses the facility within the same month, they are still charged per person. Apparently it works quite well as they make enough money to cover equipment upgrades, although not necessarily salaries.
Please do compile and send a summary of your responses to the list since this is a topic that is of continual interest to both lab managers and users. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
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Dear Listers. We are in the midst of trying to set up a new EM facility here. I am wondering how many of you out there actually support your facility with users fees? Is it realistic to expect service contracts, personnel salaries, supplies and maintenance can be recouped from fees? Or do most facilities need to be subsidized to ensure their survival? I worry that the kind of fees we would have to charge to ensure survival will scare many users away. Any suggestions, comments would be appreciated.
JoAnn Buchanan Dept. Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305
Dear Listservers, I am now looking for a resonable ultramicrotome to buy. Does anybody know where I should look for used ultramicrotomes and diamond knifes? Thank's a lot in advance.
Sincerely,
Young Ho Koh, Ph.D University of Wiconsin Madison, 445 Henry Mall Madison, WI 53706
What is the available these days in terms of Critical Point Driers. Ours seems to be on its last legs and we will possibly replace it. Manufacturers and sales welcome.
-David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford Street Halifax NS Canada B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca
Dear JoAnn, That is always the dilemma. To fully recoup the costs of running an EM facility, you would have to charge at least $200 per hour for an SEM and $400 per hour for a TEM, and hope they were booked 60% of the time. Most researchers cannot afford that. Most university EM labs will charge university researchers much less than that, but then the university must subsidise the costs. I charge researchers $25 per hour SEM and $50 per hour TEM and that covers consummables and small incidental costs, but not service contracts (I don't have any) or my salary. I get some commercial work at commercial rates, but that is less than 5% of my time. One other point, a university with a good EM facility can use that fact to attract more research funding. There is a lot of research in many diverse fields that is enhanced or facilitated by the access to modern EMs. At 04:12 PM 5/14/01 -0700, you wrote:
} } } Dear Listers. We are in the midst of trying to set up a new EM facility } here. I am wondering how many of you out there actually support your } facility with users fees? Is it realistic to expect service contracts, } personnel salaries, supplies and maintenance can be recouped from fees? Or } do most facilities need to be subsidized to ensure their survival? I worry } that the kind of fees we would have to charge to ensure survival will scare } many users away. Any suggestions, comments would be appreciated. } } JoAnn Buchanan } Dept. Molecular and Cellular Physiology } Stanford University School of Medicine } Stanford, CA 94305 } Regards and luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I have resisted diving into this issue of digitized/computer enhanced images and their use as trial exhibits as a number of good comments have been made by other well qualified scientists. For the record, I would like to note that I HAVE used digitized images and computer-processed and enhanced images (both photographs and photomicrographs) in Federal Court testimony. As several other responses to the list have noted or expressed a concern that the images might be challenged, I can assure you that they (the images) WERE questioned. Defense council objected to several computer "enhanced" or "highlighted" images which had been submitted as exhibits to support my testimony. Defense council further wanted all such images rendered inadmissible. The judge allowed me to explain to the court exactly what had been done to the questioned digital images as part of my case analysis. Though the defense council turned my phrases from "digitally recorded" and "digitally enhanced" into derogatory slurs of "digitally manipulated" and "computer-altered", the judge listened patiently to my remarks on computer digital imaging and image analysis. He then stated he could find no problem with the processes I had described and no grounds to dismiss the exhibits which he allowed to stand.
Those of us from the old "red light / dark room" days know it is very possible to "dodge" out specific regions of a photograph or "super impose" an object into a photograph which was never there in the first place. Although it took a little more time than a few keystrokes at the computer, one could achieve very convincing photographic images from the trays of the darkroom. Furthermore, in the BC (before computers) days, you seldom had to swear in court, other than your original oath, that you had not altered the images in any manner.
The credibility of the analyst / microscopist / scientist / forensic scientist/ responsible individual is the ONE THING we MUST preserve and do everything in our power to maintain.
Our credibility is the backbone of our science.
Therefore, do your scientific analysis and record ALL that you do to the evidence (i.e., images). Truthfully, report your findings and observations based upon YOUR analysis and don't "over step" your results. Be a man/woman of unquestionable character. Allow the judicial process to take it's course. (what ever course that is) THEN: To slightly paraphrase Bill Oliver's recent post, "The jury will either believe you or they won't." At that point you have done your job.
I fully realize many of the list readers are not forensic scientists and will seldom be concerned with court testimony, but permit me one last observation. Several years ago and at my request, Mr. Robert Hathaway of the Rhode Island State Crime Laboratory provided me with a copy of Brouardel's Statement about the role of the Forensic Scientist which Mr. Hathaway used in his presentation. I have had it on my office AND laboratory wall ever since. (Thanks Bob; it helps to keep things in perspective.)
" If the Law has made you a witness, remain a man of science. You have no victim to avenge, no guilty or innocent person to convict or save. You must bear testimony within the limits of science. " Dr. P.C. Boarded
Respectfully submitted,
S. Frank Platek Forensic Chemistry Center U.S. Food and Drug Administration 6751 Steger Drive Cincinnati, OH 45237-3097 (513) 679-2700 X254 (513) 679-2761 FAX fplatek-at-ora.fda.gov
Disclaimer: The opinions and views expressed above are those of author and do not necessarily represent those of the US Food and Drug Administration or any other Federal or State Agency.
I have some 5-to-8 year-olds who really enjoy the microworld, but whose hands can't keep up with speedy microbeasties under the lens. I remember from years past that there was a product (I think it was called 'Proto-Slo', or similar) that was viscous, roughly iso-osmotic, and designed to slow the critters down for easier observation. Is it still available? If not, does anyone have a recipe for a home-brewed equivalent?
Thank you, thank you, thank you,
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
We are doing immuno EM labeling experiment on formalin fixed human brain (the tissue has been fixed for more than 2 years). The labeling density is very minimal. The same antibody we have used on mice without any problem. Does anyone have experience on how to optimize the formalin fixed human tissue for immuno electron microscopy? Thank you in advance.
Xinran Liu, M.D., Ph.D. Assistant Professor Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: (214) 648-1830 Fax: (214) 648-1801 E-mail: Xinran.Liu-at-UTsouthwestern.edu
Please consider the following exciting position with ExxonMobil Chemical in Europe. If you are interested, please e-mail or contact Dr. Anton-Jan Bons in Machlen, Belgium at address anton.j.bons-at-esso.com
Please do not send replies to Mark Disko in the USA.
2-YEAR POSTDOC POSITION: TEM CHARACTERIZATION OF CATALYTIC MATERIALS
ExxonMobil Chemical Europe Inc., European Technology Center Machelen (near Brussels), Belgium
A 2-year post-doctoral research position is currently available in the Catalytic Materials Group of Basic Chemicals Technology. The group focuses on the synthesis, characterization and testing of zeolites and related materials, for application in ExxonMobil's production processes. The current position involves structural characterization by Transmission Electron Microscopy and other techniques. Facilities available in the group include TEM, FEG-SEM, EDX and XRD; a multitude of other characterization techniques is available at other ExxonMobil sites and at nearby universities. Candidates should hold a doctoral degree in physical sciences, with a strong affinity for electron microscopy, diffraction and/or crystallography. Further information can be obtained from Dr. Anton-Jan Bons, tel. +32 2 722 2838, email anton.j.bons-at-esso.com. Applications including a full CV, list of publications and name and addresses (including e-mail) of three references should be sent to ExxonMobil Chemical Europe, Human Resources Department, Hermeslaan 2, B-1831 Machelen, Belgium, as soon as possible.
Due to a rationalisation of facilities, AEA Technology is disposing of a Philips EM430 TEM/STEM.
The microscope is a 300kV instrument equipped with an Oxford Instruments EXL-II EDX system and is available with a full range of sample holders, including analytical double-tilt, heating stage, LN2 double tile cold stage and a straining stage. The instrument has been under a full service contract since new.
Ladd Research, as well as some of the other supply houses, sell CPDs. To check out ours, please go to http://www.laddresearch.com/New_Products/CPD/cpd.html
Thanks,
JD Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955
O'Neil, David wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } What is the available these days in terms of Critical Point Driers. Ours } seems to be on its last legs and we will possibly replace it. Manufacturers } and sales welcome. } } -David O'Neil } Institute for Marine Biosciences } National Research Council of Canada } 1411 Oxford Street } Halifax NS Canada B3H 3Z1 } ph. 902-426-8258 } fax 902-426-9413 } david.o'neil-at-nrc.ca
} Listers (especially those in the Southeast USA): Please check out } the details of this symposium at the web link given below. } Larry
} } Materials MicroCharacterization: Today and Tomorrow } } ASM International, Oak Ridge Chapter } Educational Symposium 2001 } } The development of new materials with novel physical and mechanical } properties will play an important role in the twenty-first century } economy. Nation-wide initiatives aimed at developing nanostructured } and other complex advanced materials have been instituted. To } develop the scientific understanding today that will tomorrow enable } the synthesis of these new classes of materials, characterization } will play a central role. The relationship between novel materials } properties and the processing methods used in their synthesis will } depend on determining the structure, composition and chemical } bonding of these materials at length scales from the atomic to the } mesoscale. This year's Educational Symposium will highlight the } state-of-the-art and future directions of a variety of materials } microcharacterization techniques. A mix of well-established and } recently developed techniques will be covered. } } The Symposium will be held at the Pollard Auditorium in Oak Ridge, } as single-day event with internationally recognized speakers from } all over the country. The date of the Symposium is Thursday, May } 31, 2001. The registration is a low $100 for professionals, and $20 } for students. The registration fee includes a morning welcoming } continental breakfast, two coffee breaks, and a catered lunch. } Registrations can be confirmed by e-mail to Cheryl Lee at } cslee-at-ornl.gov. Checks should be made out to "ASM Oak Ridge } Chapter" with "Educational Symposium 2001" noted in the memo line, } and mailed to the address below. Additional information can be found } at the web site of the ASM Oak Ridge Chapter at } http://www.korrnet.org/orcasm/. } } Speaker List: } } } Larry Allard, Oak Ridge National Laboratory } Peter Nellist, Nion Co. } Tom Kelly, University of Wisconsin } Greg Rohrer, Carnegie Mellon University } Joe Michael, Sandia National Laboratory } Harald Ade, North Carolina State University } Kent Childs, Phi Co. } Paul Carpenter, NASA Marshall Space Flight Center } David Joy, University of Tennessee
-- Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Has anyone used Oil Red O to stain lipids in tissues embedded in plastics (Epon or Epon-Araldite)? If so, has this been done by staining en bloc or by staining the sections. Sections would range from 1-4 microns in thickness.
We would also consider tissues embedded in glycol methacrylate. We'd like to avoid frozen sections because we'd prefer the higher level of detail possible with plastic.
Thank you,
Jaclynn Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Ave. St. Louis, MO 63110
FWIW, we have been using a Wacom tablet to trace outlines for image analysis. We simply used the image on the screen to guide our hand on the tablet. It really didn't take too long to develop the necessary coordination. I wonder if you would have any trouble with parallax with the LCD tablet.
The tablet has two modes of operation: typical mouse motion (relative) and screen coordinates (absolute). We had to be sure to run in screen coordinates mode to get the best performance tracing. We also had to make sure our image window was filling a large portion of the screen to get the least amount of jaggies (due to either shaking hands or limited tablet resolution) while tracing. Bigger tablets are better in that regard. We were working with a 5x7" tablet which was fairly inexpensive.
Warren S.
At 08:17 AM 5/15/2001 -0400, you wrote:
} I would appreciate from users of the Wacom PL series LCD writable tablet, } for use specifically with image analysis, not graphics arts. The ability to } trace directly on the image rather than using a mouse or pen/tablet } combination seems appropriate but would like some feedback. TIA. } } Walter F. Bobrowski } Electron Microscopy Laboratories } Drug Safety Evaluation } Pfizer Global Research and Development } 2800 Plymouth Road } Ann Arbor MI 48105
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Listies: we are preparing samples for Cu grain size analysis with a focused ion beam and then doing the analysis manually. Engineering would like to know if there is any software out there that will do this automatically or with little user intervention. Vendors can respond to me off-list.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Dear Paul, We used to use glycerin in the Microbiology lab. Only way to see Chlamydomonas. At 04:45 PM 5/15/01 EDT, you wrote:
} Dear Listers, } } I have some 5-to-8 year-olds who really enjoy the microworld, but whose hands } can't keep up with speedy microbeasties under the lens. I remember from } years past that there was a product (I think it was called 'Proto-Slo', or } similar) that was viscous, roughly iso-osmotic, and designed to slow the } critters down for easier observation. Is it still available? If not, does } anyone have a recipe for a home-brewed equivalent? } } Thank you, thank you, thank you, } } Paul Grover } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hi Warren: I am also interested, about how much are the small tablets? Thx, Mike Coviello UT Arlington
Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } FWIW, we have been using a Wacom tablet to trace outlines for image } analysis. We simply used the image on the screen to guide our hand on the } tablet. It really didn't take too long to develop the necessary } coordination. I wonder if you would have any trouble with parallax with the } LCD tablet. } } The tablet has two modes of operation: typical mouse motion (relative) and } screen coordinates (absolute). We had to be sure to run in screen } coordinates mode to get the best performance tracing. We also had to make } sure our image window was filling a large portion of the screen to get the } least amount of jaggies (due to either shaking hands or limited tablet } resolution) while tracing. Bigger tablets are better in that regard. We } were working with a 5x7" tablet which was fairly inexpensive. } } Warren S. } } At 08:17 AM 5/15/2001 -0400, you wrote: } } } I would appreciate from users of the Wacom PL series LCD writable tablet, } } for use specifically with image analysis, not graphics arts. The ability to } } trace directly on the image rather than using a mouse or pen/tablet } } combination seems appropriate but would like some feedback. TIA. } } } } Walter F. Bobrowski } } Electron Microscopy Laboratories } } Drug Safety Evaluation } } Pfizer Global Research and Development } } 2800 Plymouth Road } } Ann Arbor MI 48105 } } ---------------------- } Warren E. Straszheim } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking
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We need a 3nm colloidal gold conjugated to strepavidin. You can now purchase conjugates at 5, 10, etc in size but not 3nm so we plan to attempt to make it. I have made 5 and 10 nm conjugates using IgG's and protein-A which turned out great and do have the instructions for the 3nm colloid. I was wondering if anyone had made a strepavidin conjugate and, if so, would you share your procedure for the conjugation? I do not know if the procedure used for IgG's would work equally well for strepavidin. Thanks, Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
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Very well said, Frank.
Thanks.
Gary G.
At 11:50 AM 5/15/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am in doubt by one of our customers - specialist in image manipulations/publications. The talk is about "resolution of an digital image" - in it's virtual form - means - in a datafile format - e.g. on floppy. According to my understanding - it is described as pixel dimention of an image e.g. 1200x800 pixels + pixel digital depth e.g. 8 bit (for grayscale - 256 gray levels). This immediately determines the raw image size as uncompressed bitmap. E.g. the digital camera images resolution is defined in Megapixels defining the sensor matrix - which means - pixel X times pixel Y dimentions of image - multiplicated - independent of image ratio (rectangular, square etc.) This is understandable and clear terminology.
However the "image analysis" people,I am facing now, use the DPI or PPI parameter as a resolution measurment.
Talking about the shortcut meaning "dots per inch" or "pixels per inch" (as far as I understand it) - has only the sense when there is a specific media described/selected - monitor working at particular grafic card resolution, scanner sampling with specific optical sensor resolution, hardcopy (printout, photo, negative or videoprint) having specific size and printing head resolution....etc. This "inch" must be defined somwhere in physical way.
Some people told me that ANY image in datafile format has own RESOLUTION defined in this DPI's or PPI's... ????? How this should be understood ???
It is for me an absurd until one specifies the physical media where this "inch" is defined.
PLEASE advise with Yr experience - is that right or not - how the resolution specified in DPI should be related to image pixel per pixel resolution.
kind regards, awaiting clarification....
Krzysztof Herman EMISJA - FEI EO Poland ul.Ba¿ancia 45A 02-892 Warszawa tel/fax: (+48 22)6449753, 6449750 mobile: (+48 601)307456 kherman-at-labsoft.com.pl www.emission.com.pl
MSA Listserver is a good source for used ultramicrotomes, but you'll have to do some searching.
Microscopy Today magazine has a used equipment for sale column toward the back. See http://www.microscopy-today.com.
Used diamond knives: It would cost a couple thousand to resharpen used ones. You might try Microstar Technologies in Texas for new ones: phone 409-291-6891, email: mistar-at-msn.com for a catalog.
I'm not affiliated with those enterprises.
Greg -- Greg Lum Computer/Microscopy Consultant Dept of Biology San Francisco State University Ph: 415/338-1339 email: glum-at-sfsu.edu
A friend is looking for a low cost inverted light microscope (ca $15,000). I am not very knowledgeable about what is available so look to you for advice.
She was particularly interested in something equivalent to the Zeiss Axiovert 100, 135, or even better, the 100TV.
I do know these are no longer being supplied by Zeiss so the names of reliable sources for used microscopes would be useful.
All information supplied will remain confidential, although I may post a list of useful contacts back onto this forum.
Regards,
Paul Webster
Paul Webster, Ph.D. Scientist II and Director Ahmanson Center for Advanced EM & Imaging House Ear Institute 2100 West 3rd Street Los Angeles CA 90057
} Some people told me that ANY image in datafile format has own RESOLUTION } defined in this DPI's or PPI's... ????? } How this should be understood ???
Some image formats include such information in the file. This allows one to have an idea about the physical dimensions of the original object. If only the value of DPI or PPI is specified for an image then it is not possible to relate it to image pixel resolution. Both DPI and size of the image in inches should be specified, then:
Krzystzof You have defined two aspects of "resolution" in your question, (1) pixel resolution relating to the sensor resolution) (2) pixels per inch on the display or printer output. A third is pixels per inch of real-world dimensions of the object being imaged. This is the spatial calibration or spatial resolution, and must be known if the sizes of objects in the image are to be measured by image analysis.
When you scan a paper image (e.g. a photographic print) on a flat-bed scanner into e.g. Adobe Photoshop, Photoshop records pixels x,y and pixels per inch in the file header. Then when the image is printed it emerges from the printer the same size as the original. In this particular case, resolution types 2 and 3 are initially the same. But both pixels x,y and ppi can be altered using Photoshop's image size dialog. If either are changed then the relationship of Type 1 and Type 2 resolution to Type 3 resolution is changed, and it will no longer be possible to determine an object's dimensions. If a digital camera or microscope image of a real-world scene is captured via a Twain grabber into Photoshop, Photoshop will report pixels x,y and also pixels per inch for such an image. It seems to define ppi by scaling the input pixels x,y into the currently-defined page, so 768x576 pixels comes out as 72 ppi on A4 paper. But the type 3 resolution, real-world pixels per inch of objects in the image, cannot be calculated until some object of known dimensions is imaged under the same conditions.
The problem here is that we tend to use the same word "resolution" to mean any of these things without specifying which. We should perhaps use a second qualifying word such as sensor resolution, pixel resolution, image resolution, print resolution, spatial resolution. This helps, but you can see from these examples that it is hard to avoid ambiguity.
Chris
} ----- Original Message ----- } From: "Krzysztof Herman" {kherman-at-labsoft.com.pl} } To: "MSA" {Microscopy-at-sparc5.microscopy.com} } Sent: Thursday, May 17, 2001 12:21 AM } Subject: image resolution... } } } -------------------------------------------------------------------- -- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------- -- } -. } } } Hello Listservers } } I am in doubt by one of our customers - specialist in image } manipulations/publications. } The talk is about "resolution of an digital image" - in it's virtual } form - } means - in a datafile format - e.g. on floppy. } According to my understanding - it is described as pixel dimention of } an } image e.g. 1200x800 pixels + pixel digital depth e.g. 8 bit (for } grayscale - } 256 gray levels). } This immediately determines the raw image size as uncompressed } bitmap. } E.g. the digital camera images resolution is defined in Megapixels } defining } the sensor matrix - which means - pixel X times pixel Y dimentions of } image - multiplicated - independent of image ratio (rectangular, } square } etc.) This is understandable and clear terminology. } } However the "image analysis" people,I am facing now, use the DPI or } PPI } parameter as a resolution measurment. } } Talking about the shortcut meaning "dots per inch" or "pixels per } inch" (as } far as I understand it) - has only the sense when there is a specific } media } described/selected - monitor working at particular grafic card } resolution, } scanner sampling with specific optical sensor resolution, hardcopy } (printout, photo, negative or videoprint) having specific size and } printing } head resolution....etc. } This "inch" must be defined somwhere in physical way. } } Some people told me that ANY image in datafile format has own } RESOLUTION } defined in this DPI's or PPI's... ????? } How this should be understood ??? } } It is for me an absurd until one specifies the physical media where } this } "inch" is defined. } } PLEASE advise with Yr experience - is that right or not - how the } resolution } specified in DPI should be related to image pixel per pixel } resolution. } } kind regards, awaiting clarification.... } } Krzysztof Herman } EMISJA - FEI EO Poland } ul.Ba¿ancia 45A } 02-892 Warszawa } tel/fax: (+48 22)6449753, 6449750 } mobile: (+48 601)307456 } kherman-at-labsoft.com.pl } www.emission.com.pl } } } }
We have an Americium x-ray source that we no longer need. My understanding is that disposal of this is extremely expensive. Has anyone dealt with this problem and can give me some suggestions? Thank you, Lynne C. Garone Polaroid Corp. 1265 Main St. W4-1D Waltham, MA 02451-1714 (781) 386-1446 GaroneL-at-Polaroid.com
Below is the result of your feedback form. It was submitted by (yarrabee-at-caboolture.net.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 17, 2001 at 04:16:18 ---------------------------------------------------------------------------
Email: yarrabee-at-caboolture.net.au Name: eliza
Organization: QUT
Education: Undergraduate College
Location: brisbane, qld, australia
Question: hello. i am doing an assignment for an EM subject at uni, and was wondering if anyone could steer me in the right direction. my question is about which set of methods i would use to prepare rat liver cells for the examination of mitochondrial ultrastructure. would it be fair to assume that the usual methods of sample preparation for TEM could be used in this scenario? that is, dehydration, fixation and embedding; with the protocols optimised for mitochondrial preservation? i was also wondering which if any fixative additives would be appropriate in this case, eg. tannic acid/potassium ferrocyanide. and whether any immunolocalisation specific to mitochondria would be applicable to this case. thank you for your help. eliza.
As the lead software developer for ASPEX's Personal SEM product, I have had opportunity to consider this issue. I've come to the conclusion that the crux of the problem is sloppy use of language. The word "resolution" is used in (at least) two different contexts - resolution to mean "pixel dimensions" (i.e. 1024 x 768) or resulution meaning "pixel size". Within the "pixel size" meaning there is additional ambiguity as to whether "pixel size" refers to the edge length on the *sample* as represented by a single pixel or whether "pixel size" refers to the edge length on the *display surface* (printer, monitor etc.). Typically image formats refer to the edge length of a pixel on the display surface. Many (but not all) image formats have pixel size (display surface related) data encoded with the image. This datum is just a suggestion and usually (in microscopy) does not have deep or profound meaning. Usually when the image is displayed on a video monitor, the image is displayed by mapping one display pixel to one image pixel thus overriding the suggested "pixel size" with the default ~96 to 128 dpi typical of most monitors. There is one situation in which the "pixel size" can represent more than a suggestion. If the image has a magnification marker burned into the image, the magnification marker *may* be strictly correct only when the image is displayed at the suggested pixel size. By strictly correct I mean that the magnification is defined as the scale factor between the length of a pixel edge on the sample to the length of a pixel edge on the display surface. Of course, this is not an issue if you use micron bars rather than magnification and since digital images are so easily rescaled I always recommend the use of micron bars over magnification.
Hope this helps,
Nicholas ############################### # Nicholas W. M. Ritchie # # Aspex Instruments # # 175 Sheffield Drive # # Delmont, PA 15626 # # (724) 468-5400 # # nritchie-at-aspexllc.com # ###############################
We have a software package (analySIS) from Soft Imaging System in Colorado that includes grain size analysis per ASTM specs. We use it for both SEM and LM images and have been happy with it. It allows you to manipulate your image so the grain boundaries are well defined.
Caveat: I have no involvement with SIS other than being a pleased customer.
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 jlagoy-at-bodycote-imt.com
} } Hello Listers, } I was wondering if anyone out there can help me out. I am looking } for a solution of uniform particles (spheres would be nice) of a heavy } material so that it will show up nicely in backscatter. I also require that } the particle size be between 2 and 5 micron. If anyone knows of something } like this and where I can order it from, It would help me out. } Thanks in advance } Nick
Details for submission of short Contributed Talks, Registration Form and Accommodation info for the Oxford EFTEM Meeting are to be found now on the NEW Royal Microscopical Society website :
http://www.rms.org.uk (+details below) under the EVENTS link.
LT
**************************************************************************** ********* DEVELOPMENTS IN ENERGY-FILTERED ELECTRON MICROSCOPY
A meeting organised by the Royal Microscopical Society and supported by EMAG, FEI UK Ltd, JEOL (UK) Ltd., LEO EM Inc., Gatan UK, TVIPS GmbH
Department of Materials, Oxford University
Wednesday 4 July 2001
REGISTRATION Online printable form at http://www.rms.org.uk/current%20events.html#devenergy
CONTRIBUTED TALKS Contributed presentations are now being sought. Abstracts of no more than 200 words should be sent by email to: crispin.hetherington-at-materials.ox.ac.uk and should arrive by 31 May, 2001.
OVERVIEW and INVITED SPEAKERS The use of electron energy filters in analytical electron microscopy (EFTEM) is a relatively recent development that is proving to be an extremely powerful tool. In-column filters (e.g. so-called "omega-filters") and post-column filters (e.g. the "GIF") represent different approaches which have their own advantages as well as experimental difficulties. This one day meeting is designed to explore the current state of the art, both with regard to the instrumentation and also applications of EFTEM in life and physical sciences.
Dr Bernd Feja (Tietz Video & Image Processing Systems GbmH) Energy-filtered electron tomography Prof Joachim Mayer (Aachen University of Technology) EFTEM - the state of the art and future trends Dr Paul Midgley (University of Cambridge) EFTEM image series - taking elemental mapping into a new dimension Prof Michael Trendelenburg (German Cancer Research Center, DKFZ) EFTEM in biomedicine & biotechnology: Recent advances in specific element mapping
Note this meeting is timed to follow immediately the 1-day FEGTEM meeting to be held on 3 July 2001, also in Oxford.
FURTHER DETAILS Dr Crispin Hetherington, tel. 01865 273799, crispin.hetherington-at-materials.ox.ac.uk Dr Laurence Tetley, tel. 0141 330 4431, l.tetley-at-bio.gla.ac.uk
ACCOMMODATION Accommodation in Oxford is listed at http://www.oxfordcity.co.uk/accom. The top page lists the hotels in the £60 and £100 range, but there are links to many cheaper places. For students (and others?) there is also a brand new Oxford YHA at 2a Botley Rd, right next to the station. 184 beds at £18 (B&B). Tel 01865 762997 or try online reservations on http://www.yha.org.uk. **************************************************************************** **************** Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
} } Hello Listers, } I was wondering if anyone out there can help me out. I am looking } for a solution of uniform particles (spheres would be nice) of a heavy } material so that it will show up nicely in backscatter. I also require that } the particle size be between 2 and 5 micron. If anyone knows of something } like this and where I can order it from, It would help me out. } Thanks in advance } Nick
This is indeed a source of confusion. "DPI" or "PPI" makes sense if you want to print something, as the printers themselves have a given resolution. For example, if you send something to a printer which has 300 DPI, it is not very useful to send an image with 1200 DPI. The details don't get printed and you are wasting space (and possibly bandwidth). So, you are right, that the medium palys a role for this measure. The other use of this value comes from scanning (negatives or photos), because again the resolution is given by the scanner in "DPI". For any microscopy work or image analysis, the values of "DPI" have little meaning, as it is the CONTENT of the image that is of interest, not the MEDIUM. Let's say you take an image in a TEM at 1,000,000x mag. On a ngative (about 10 cm), you would see roughly 10^-7 m or sample, or 100 nm. If you take a digital camera and take the same area with a roughly 1Kx1K resolution, you'd have a resolution of 2.5 x 10^8 DPI, if you wanted to refer to the real world length coordinates, clearly nonsense. However, if you printed the image on paper with a print size of 10 cm (4 inch), you would have a resolution of 250 DPI (1000 dots / 4 inch), which is a value that fits a printer resolution well.
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Radostin Danev [mailto:rado-at-nips.ac.jp] Sent: Wednesday, May 16, 2001 11:44 PM To: MSA
Dear Krzysztof,
} Some people told me that ANY image in datafile format has own RESOLUTION } defined in this DPI's or PPI's... ????? } How this should be understood ???
Some image formats include such information in the file. This allows one to have an idea about the physical dimensions of the original object. If only the value of DPI or PPI is specified for an image then it is not possible to relate it to image pixel resolution. Both DPI and size of the image in inches should be specified, then:
Dear Microscopists: } } Dr. Charles Francis of the Vascular Medicine Unit is listing for sale two EM } microscopes and other equipment from an EM lab which will be shutdown this } summer. The equipment and microscopes are at the University of Rochester } Medical Center in Rochester, NY (upstate NY). } } 1) 1989 JEOL JSM-T330A Digital Scanning EM } } 2) 1983 Zeiss EM10C Transmission EM } } Also, auxillary equipment, such as: } } Water chiller: Coldwell } } Reichert TM 60 specimen trimmer } } Two Reichert OMUS Ultramicrotomes, purchased in the late 1970's & early 80's } and still functioning well. } } LKB Historange 2218 microtome } } If you are interested in these, please e-mail } } Administrator, Elizabeth Corrigan: beth_corrigan-at-urmc.rochester.edu } } OR } } EM technical consultant, Karen Jensen, M.S.: } karen_jensen-at-urmc.rochester.edu }
I've run into this problem before, its the confusing terminology used by printers and graphic artists. In a digital microscope image the resolution is the size of the pixel in microns, everything else is related to output (screen or paper). See the following links that I took from my web page at (note that some of the URLs may wrap to a second line): http://swehsc.pharmacy.arizona.edu/exppath/micro/digimagehardware.html
http://www.kodak.com/US/en/digital/dlc/book3/chapter5/lesson1/p01.shtml Image Resolution (Kodak - Printing Digital Images)
http://graphicdesign.about.com/arts/graphicdesign/library/weekly/aa070998.htm Resolution: DPI, SPI, LPI and PPI (About.com)
http://www.adobe.com/support/techguides/printpublishing/scanning/psscanning.html Introduction to Halftones & Scanning (Adobe)
Best regards. Doug
At 01:21 AM 5/17/2001 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
.................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : :...................................................................: http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
} However the "image analysis" people, I am facing now, use } the DPI or PPI parameter as a resolution measurement. } } ... } This "inch" must be defined somewhere in physical way. } } Some people told me that ANY image in datafile format has } own RESOLUTION defined in this DPI's or PPI's... ????? } How this should be understood ??? } } It is for me an absurd until one specifies the physical } media where this "inch" is defined.
I believe the "image analysis" people you refer to speak of a different context of resolution, different from the definition of "resolution" which is common to image file formats. This latter definition is simply a preference for printing which can be changed at any time. Using your 1200 x 800 example, you could print the image at 6" by 4" at 200ppi, or print it at 12" by 8" at 100ppi ... neither one of these "print" definitions change the pixel bitmap, rather instruct the printer where on paper to put the pixel.
"Image analysis" does require a different definition for resolution which belongs to the object which is imaged ... and this definition should not change unless the original number of pixels is changed ... for example, if the original 1200 x 800 is "resampled" to 600 by 400. Image analysis might want to know the magnification of the image ... better, know the "real dimension" of a pixel, or the what the distance between pixel represents in reality. Does it represent 1 micron, 2.6 microns, etc. If this information is known, then image analysis software can calculate linear measurements or determine areas and present the information in real terms ... microns, centimeters, kilometers ... rather than pixels or dots.
As far as I know, no common image format (e.g., TIF) has a place for defining the "image analysis" context of resolution, only the "printer" definition. Rather, image analysis software needs to be instructed of what the real dimensions of a pixel are. There are softwares which will write "real" resolution definitions to a TIF file, but is would be uncommon if a different software would know where to find it. This is the case for my SEM software ... it knows where to find the magnification definition, but my Image Pro software does not.
any help? ... cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
Am I the only one not receiving anything from the listserver. I didn't unsubscribe. I only got stuff from the ICEM folks.
Paula :-(
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
I am using the Leica 2168 Ultrostainer in a cost recovery facility. Where it cost my users $4.25 per stain run a year ago, I will now have to charge them $21.00 per run, due to an unbelievable increase in price and reduction of quantity for supplies.
Does anyone have any suggestions regarding reducing the cost to my users, such as making stain and refilling bags, other suppliers of stain, or manufacturers of automatic stainers, etc.
Kathy Troughton Senior Research Technician University of Tennessee, Memphis
Dear Paul, If you can get your hands on a Bid Service catalogue, it will usually have a few inverted light microscopes. Bid Service is a supplier of used semiconductor manufacturing equipment. Their prices are usually too high, but they are worth a visit. www.bidservice.com At 05:27 PM 5/16/01 -0700, you wrote: } } Dear All, } } A friend is looking for a low cost inverted light microscope (ca $15,000). } I am not very knowledgeable about what is available so look to you for } advice. } } She was particularly interested in something equivalent to the Zeiss } Axiovert 100, 135, or even better, the 100TV. } } I do know these are no longer being supplied by Zeiss so the names of } reliable sources for used microscopes would be useful. } } All information supplied will remain confidential, although I may post a } list of useful contacts back onto this forum. } } } Regards, } } Paul Webster } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
you will probably get lots of answers to this but here goes: the value in pixels is a simple quantity (like distance) whereas dpi or ppi is a rate (like velocity) so they cannot be compared directly unless you can multiply the rate by inches (or in velocity by time).
You have already said that the importance of dpi is related to a physical medium. So the two important physical media are source or object originally captured and the output medium (printer, display screen etc.). If your 1200 dpi camera can focus close enough to photograph something 1 inch long then its best resolution would be 1200 dpi or the smallest thing you could see would be 1/1200 inch (approx. 20um). If you use an SEM with digital capture then it's a simple matter to calculate the digital part of it's resolution for a particular object which is photographed. The point in both cases is that the resolution changes because you can get closer or further away. But if you capture an image using a scanner then the maximum resolution (e.g. 1200 dpi is fixed).
However if you want to output this to a photographic print you may be more interested in the maximum size of print you could create. This is affected by a variety of considerations but the simplest one is to produce a picture size where the maximum detail can be shown without loss of resolution. The average human eye at optimum viewing distance can resolve about 0.2mm so the maximum good print should measure 1200x0.2mm by 800x0.2mm (i.e. 240mm x 160mm or approx. 9 x 6 inches). If you want to make a bigger picture with maximum resolution then you need a camera with more pixels, if you just want to show more detail in your sample you could keep the picture at the same size but photograph the sample closer.
So, if you are printing, the resolution of your image must be at least about 150 dpi or for screen output about 75 dpi for the average display monitor. The number of pixels required would then be dictated by how big your picture would be on screen or paper. But you might also need to consider that your audience may want to enlarge the image on screen, view a print with a magnifier or stand further back to view a display picture.
This whole situation is made worse, of course, by claimed resolutions of printers. If it's an inkjet or laser printer then the claim of 1200 dpi for output is nearer to 20um (or less depending on whose figures you use) because dithering of several ink spots is required to produce shades of grey or tones of colour whereas dye sublimation and image printers usually achieve their actual figure claimed figures of 200 or 300 dpi. So there's no point in creating an image with 400 dpi onto an inkjet print because the maximum that could be represented by the printer would be about 200 dpi.
I hope this helps.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Krzysztof Herman wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Listservers } } I am in doubt by one of our customers - specialist in image } manipulations/publications. } The talk is about "resolution of an digital image" - in it's virtual form - } means - in a datafile format - e.g. on floppy. } According to my understanding - it is described as pixel dimention of an } image e.g. 1200x800 pixels + pixel digital depth e.g. 8 bit (for grayscale - } 256 gray levels). } This immediately determines the raw image size as uncompressed bitmap. } E.g. the digital camera images resolution is defined in Megapixels defining } the sensor matrix - which means - pixel X times pixel Y dimentions of } image - multiplicated - independent of image ratio (rectangular, square } etc.) This is understandable and clear terminology. } } However the "image analysis" people,I am facing now, use the DPI or PPI } parameter as a resolution measurment. } } Talking about the shortcut meaning "dots per inch" or "pixels per inch" (as } far as I understand it) - has only the sense when there is a specific media } described/selected - monitor working at particular grafic card resolution, } scanner sampling with specific optical sensor resolution, hardcopy } (printout, photo, negative or videoprint) having specific size and printing } head resolution....etc. } This "inch" must be defined somwhere in physical way. } } Some people told me that ANY image in datafile format has own RESOLUTION } defined in this DPI's or PPI's... ????? } How this should be understood ??? } } It is for me an absurd until one specifies the physical media where this } "inch" is defined. } } PLEASE advise with Yr experience - is that right or not - how the resolution } specified in DPI should be related to image pixel per pixel resolution. } } kind regards, awaiting clarification.... } } Krzysztof Herman } EMISJA - FEI EO Poland } ul.Ba¿ancia 45A } 02-892 Warszawa } tel/fax: (+48 22)6449753, 6449750 } mobile: (+48 601)307456 } kherman-at-labsoft.com.pl } www.emission.com.pl
I am trying to locate author information for the Journal of Electron Microscopy Techniques but I can't find a thing on the web. Does such a journal exist or is my mind confused? I swear I saw such a journal once.
Bob Robert R. Wise, Ph.D. Associate Professor of Plant Physiology Department of Biology and Microbiology University of Wisconsin Oshkosh Oshkosh, WI 54901 tele: (920) 424-3404 fax: (920) 424-1101 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
I am looking for a confocal microscope in the Dallas TX area that I could use. I have experience with TEM, SEM, light and standard fluorescent microscopy, but only minimal experience with confocal.
I am planning on submitting proposals for some immunohistochemistry work on inner ear tissue where I feel confocal imaging of intact tissue would yield more information that standard cross section analyses. If anyone has info on who might have a system in the Dallas area that would allow outside users or on how much it would cost to use, please contact me by my direct e-mail.
Thank you,
Karen Pawlowski, Ph.D. University of Texas at Dallas
It seems to me I remember seeing little spheres for sale in the microscopy supply house catalogues a few years ago. They were made in space, on the Shuttle, I believe, in zero gravity so were guaranteed to be perfectly round. I'm not sure of their sizes or composition (they may well be too light for your purposes). They may even have been sold as "Space Microspheres" (or "Space Balls?" - or was that a movie?)...They say the memory is the second thing that goes......
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
----- Original Message ----- } From: "Nicol Aitken" {nicol-at-semiconductor.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 17, 2001 11:30 AM
I am trying to locate author information for the Journal of Electron Microscopy Techniques but I can't find a thing on the web. Does such a journal exist or is my mind confused? I swear I saw such a journal once.
Bob
Dear Bob, The journal was renamed some years ago; I think the new name was Microscopy Research and Technique(s). I think it still exists. Yours, Bill Tivol
The name of the journal was changed to Microscopy Research & Technique. http://www.interscience.wiley.com/jpages/1059-910X/
} } } I am trying to locate author information for the Journal of } Electron Microscopy Techniques but I can't find a thing on the web. Does } such a journal exist or is my mind confused? I swear I saw such a journal } once. } } Bob } } Dear Bob, } The journal was renamed some years ago; I think the new name } was Microscopy } Research and Technique(s). I think it still exists. } Yours, } Bill Tivol
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
In a message dated 5/17/01 2:13:23 PM, wise-at-vaxa.cis.uwosh.edu writes:
} I am trying to locate author information for the Journal of } Electron Microscopy Techniques but I can't find a thing on the web. Does } such a journal exist or is my mind confused? I swear I saw such a journal } once.
It is now Microscopy Research and Technique, very much alive. Contact Dr.John E. Johnson, Jr., the Editor in Chief, at (650) 366 1644 or email JEJ-at-Neuroscience.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Subject: TEM- cell colture fixation and embedding } } I need to study the ultrastructural aspects of a cell line grown on petri } dishes. Any suggestion for the best fixation and embedding protocol (no } pellets, just fixation and embedding in the dish itself) ? } } thanks
I have used Epon substitutes for this with good results. Spurr's resin reacts with the plastic dish and must be avoided. I fixed, rinsed, osmicated, rinsed as usual. Dehydration with graded ethanols. Skip the propylene oxide, it dissolves the dish. Epon will mix with ethanol well enough, I used a graded series of 2:1, 1:1, 1:2 ethanol:Epon followed by at least 2 changes of pure Epon. Frequent agitation in all of the steps containing Epon to insure removal of solvents. Polymerize as usual. Find an area on the dish you are interested in. Now get a scalpel blade and heat it in a flame. Use it to cut through the both plastic dish and the Epon. Often the stress of the cutting will separate the Epon from the dish or bit of force will separate the two. Use a file to flatten and roughen the surface of Epon away from the cells. Glue the small piece of Epon with the cells of interest to a blank Epon block with "Super Glue". Cut sections. Remember if you are cutting en face, parallel to the surface the cells were grown on, you will have very little material to section before you go all the way through your specimen and hit pure Epon.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I want to thank everyone who responded to my question about whether user fees could cover service facility charges. The answer was a resounding "NO". I received responses from 20 people, 15 of whom were facility managers across the country. All labs required subsidy from their University or individual departments. The kind of charges it would take to cover all costs would be about $200.00 per hour for SEM and $400.00 per hour for TEM and 60% usage!!. Users object to paying high prices for services, so business drops off, and the facility shuts down. The burden is best shared among departments and should involve a combination of sources including research grants, user fees, and institutional support. At best, service fees cover 30-50% of costs associated with a facility. This will give me the necessary information to (hopefully) convince the powers that be to give us some money. Otherwise, I guess the new EM will have to go in the garage.
JoAnn Buchanan Dept. Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Frank: Those space beads (SRM-1960 size of 10um, and SRM-1961 size of 30um) are latex particles produced by the NASA during the Challenger STS-6 and STS-11 missions, respectively.
At 03:24 PM 5/17/01 -0300, Frank Thomas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Joseph Fu National Institute of Standards & Technology 100 Bureau drive Stop 8212 Gaithersburg, MD. 20899-8212 Tel: 301-975-3495 Fax: 301-869-0822 Email: jofu-at-nist.gov
F. C. Thomas wrote: ======================================================== It seems to me I remember seeing little spheres for sale in the microscopy supply house catalogues a few years ago. They were made in space, on the Shuttle, I believe, in zero gravity so were guaranteed to be perfectly round . I'm not sure of their sizes or composition (they may well be too light for your purposes). They may even have been sold as "Space Microspheres" (or "Space Balls?" - or was that a movie?)...They say the memory is the second thing that goes...... ========================================================= You have a very good memory.
SPI Supplies placed an order for these made-in-space microspheres (in the late 1970's) when the program was first announced, and we had the distinction of being the world's first purchaser of something actually made in space! I am not sure that got us anything other than a feature one night on the Nightly News. But despite all the publicity, at the time, that was given out by NASA, I am unaware of anyone who ever published anything showing that the "perfectly round" aspects of these microspheres (or any other aspect for that matter) was anything above and beyond what is routinely made on earth! We ourselves could not find that superiority, but perhaps someone better that ourselves did succeed in finding such a difference.
The last I heard about that was that someone at Lehigh University had some funding (remember we are talking about the early 1980's) to try to find that these spheres did, in some way go beyond what could be made on earth, but I never heard anything after that.
BTW, we subdivided the vial purchased form NASA, and advertised them (as we were encouraged to do) and over the years, as I met people at our exhibit booth at trade shows, I never encountered anyone who found, when examining these spheres, anything "above and beyond" earth made spheres. When the set of samples was sold off, we never replenished our stock because we could not in good conscience tell customers that the far higher price could be justified in terms of there really being something special.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
This is a weird question, it could only happen in Santa Cruz.
One of our EH&S people stopped by today to ask if we could analyze some dust from one of the science buildings on campus. I said 'Maybe' and listened to the rest of the story.
He told me that a new custodial employee was afraid that the dust in one of the science buildings was full of toxic material and wanted it tested. The building is a 30 year old structure that has had various sorts of biology and chemistry teaching and research labs. As far as I know there has never been any sort of problem there and EH&S keeps track of things like radiation and highly toxic situations.
I was reluctant to get involved. Our EDS could probably see some things and I could recognize some mold spores and cotton fibers, but who am I? I don't have any training or certification in toxic dust analysis. Plus I probably already think this guy is a little paranoid about dust, so I probably wouldn't find anything even if it was there.
Are there labs that can do this sort of analysis and can certify that there is nothing toxic to worry about in a sample of dust? How does one sample for these things and how do we respond to this kind of request. What are the odds that the dust is anything more than just dust? What is dust anyway?
I promised our EH&S guy that I would ask around among the smartest group of folks I know. So what do you think?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I am posting this for Ricardo Drut. Ricardo wants to know if anyone has used Tissue -Tek Tissue-Clear as a substite for Xylol, and how do you like it. You may respond to Ricardo at his email address. Thanks, Teresa
} ¿Alguien usa este producto como aclarante sustituto del xilol? ¿Con qué } resultados?
I'm wondering who would like to share a day to day consumable list for a multiple user EM (both TEM and SEM, and EDS and GIF) lab in materials science. Idealy, I would like to see a relatively exhaustive but still common consumable list with both the users (sample prep/imaging) and the service/maintenance in mind. Thanks in advance!
Chaoying Ni Director of Electron Microscopy Lab Dept. of Materials sci. and Eng. University of Delaware Newark, DE 19716 cni-at-udel.edu
I would like to get an idea about the way to look at a morphology of a nano sized powder sample in TEM.
I need to take TEM morphology pictures from MgO powder (from nano meter to a few micron size distribution) using a Cu grid. To spread a MgO particle on the grid, I will spray some kinds of alcohol on the powder which resides on the grid. Here, I would like to know what kinds of alcohol can be used to get a clear image without melting the holey carbon film on the grid.
I appreciate your comments.
Jae
******************************** Jae-yong Kim Post-doctoral Research Associate Chemistry Department Brookhaven National Laboratory Upton, NY 11973 Tel:631-344-4317 Fax:631-344-5815 ********************************
Is the dust toxic? Interesting question from the technophobic world. Beyond looking for the obvious heavy metals and suspicious asbestos like fibers, a microscopist/EDS analyst would have a hard time answering that question. You would have to treat the dust like a new drug to answer the question. Feed large quantities to mice and stick it into rabbit eyeballs to look for toxic symptoms and death. But don’t let the animal rights people know that you are doing such torture to animals. It is better that people get sick than to make animals suffer. UC Santa Cruz probably has a policy against such testing anyway.
Ron Vane XEI Scientific
-----Original Message----- } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Your particle size is a bit large for "colloidal" gold. How about magnetite spheres - as described below? Certainly they should show up in backscatter. My only concern are the on/off magnetic properties. I guess that an SEM (or optical scope!) would not be affected, but in TEM, with the powerful objective lens right next to the particles, there would be a problem. Those particles are available in graded and defined sizes from 1 to 8um. Disclaimer: ProSciTech is a world-wide distributor for Spherotech, but it makes little sense for US user to come to us. I suggest that you check out the spherotech.com site. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
PARAMAGNETIC PARTICLES - MAGNETIC PARTICLES The magnetic particles are prepared according to the procedures described in US Patent No. 5,091,206. They are prepared by coating a layer of magnetite and polystyrene onto monodispersed (ie. uniform sized) polystyrene core particles. As a result, the magnetic particles are spherical in shape, and paramagnetic in nature. They are also very uniform in size. The magnetite contents of these magnetic particles can be adjusted but in general it represents about 10% to 15%. The magnetic particles can be easily separated from a suspension magnetically. These particles become non-magnetic when removed from a magnet, and do not retain any detectable magnetism even after repeated exposure to strong magnetic field. The magnetic particles can be used for cell separation, affinity purification, DNA probe assays, magnetic particle EIA, etc.
On Thursday, May 17, 2001 11:54 PM, Nicol Aitken [SMTP:nicol-at-semiconductor.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } Hello Listers, } } I was wondering if anyone out there can help me out. I am looking } } for a solution of uniform particles (spheres would be nice) of a heavy } } material so that it will show up nicely in backscatter. I also require } that } } the particle size be between 2 and 5 micron. If anyone knows of something } } like this and where I can order it from, It would help me out. } } Thanks in advance } } Nick
Those were sold by NIST, I believe, and were latex spheres. Latex spheres are still available in a variety of sizes, but the special property of the space grown variety was the small standard deviation of size resulting from their microgravity production. There was only one or two batches made, prior to the Challenger incident, and all were probably sold long ago.
Latex would not be a good choice, obviously, but there are gold microspheres made for antibody research that may work. I, unfortunately, don't know what sizes they are produced in or the distributors who might have them off the top of my head. However, pump a search engine with "colloidal gold" as a search phrase and you should find them all.
On Thursday, May 17, 2001 1:24 PM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] wrote: } } Nicol - } } It seems to me I remember seeing little spheres for sale in the microscopy } supply house catalogues a few years ago. They were made in space, on the } Shuttle, I believe, in zero gravity so were guaranteed to be perfectly } round. I'm not sure of their sizes or composition (they may well be too } light for your purposes). They may even have been sold as "Space } Microspheres" (or "Space Balls?" - or was that a movie?)...They say the } memory is the second thing that goes...... } } F.C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada } } ----- Original Message ----- } } From: "Nicol Aitken" {nicol-at-semiconductor.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Thursday, May 17, 2001 11:30 AM } Subject: Microspheres } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- ---. } } } } } } } } } } } } } } Hello Listers, } } } I was wondering if anyone out there can help me out. I am looking } } } for a solution of uniform particles (spheres would be nice) of a heavy } } } material so that it will show up nicely in backscatter. I also require } } that } } } the particle size be between 2 and 5 micron. If anyone knows of } something } } } like this and where I can order it from, It would help me out. } } } Thanks in advance } } } Nick } } } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
You think the question is strange? Be careful what you look for - you may find more than you want.
Most probably missed it, but ABC recently did a piece in their national network news about St. Charles East High School, the school my younger daughter is a junior in. For years there have been complaints of air problems in the school related to symptoms of allergic problems of all kinds. The school has tried numerous labs to try to understand what may be the cause. Well and good, my own daughter has had problems that we can directly link to her time in that school, and I'd like to know why.
During spring break this year, they brought in one company from out of state. This company discovered several molds growing behind the walls that caused the school district to just shut the whole school down. The students (over 3000 of them), after a few weeks off, are now time sharing facilities with a second high school in town while the school board decides if they will have to tear the buildings down or just remodel.
In the last week, the same molds were found infesting a number of buildings and homes in Texas. The buildings have been condemned until the problem is taken care of and several home owners are finding that it will cost them more than $50,000 just to get back into their houses.
Stachybotris(?) is the culprit. There are few studies on its effects that I am aware of, but the common assumption is that it is dangerous, if not deadly. I recall something around a year ago that pointed to the possibility of potentially serious effects on infants exposed to it. Yet the current student population had an average of 6000 student years of possible exposure without a serious link, but they can now not be exposed for a second more. In fact, after weeks of debate, the school reluctantly allowed parents in for a limited time to collect anything left in school and gym lockers, after signing legal waivers and being checked in and out (I did not appreciate collecting gym clothes after weeks of fermentation).
On a similar vein, in the Chicago area, there has been considerable sensitivity to mercury. Several contractors hired by the local electrical utility apparently screwed up the replacement of mercury containing usage meters. It got to the point where the utility was doing forced testing of any home that may have been affected - if any trace of mercury was found, expensive remediation efforts were required before the homeowners were allowed back in.
One poor dope, in this time frame, uncertain what to do after breaking a mercury thermometer at his place of work, decided to call the local fire department. The building was immediately shutdown and quarantined by the EPA and the business had to wait, and pay for, space-suited remediation teams to clear all traces of mercury from the building.
In the case of both contaminants, potential levels of exposure were not a consideration. Simply finding one of these contaminants resulted in the unrealistic total quarantine response. I'm sure that these responses are not isolated in today's litigious society, just not documented as a whole.
Any building built 30 - 40 years ago was probably of a certain 'energy efficient' design of the time. In other words, windows don't open and fresh air don't get in. While reducing the loss of heated or cooled air, and thus producing less of a load on HVAC designs, these buildings essentially ensure that anything entering their environment will stay there. That includes moisture, which promotes bacterial, mold and fungus growth, as well as any material introduced. 'Sick Building Syndrome' is now a classified problem that covers more than you can imagine - including the release of gaseous compounds from new carpeting, upholstery, wood and plastic materials as well as any seemingly minor release of any solid, liquid or gaseous material that ever happened in the building.
Take my advice for what you paid - let the EH&S people find their own resources. This seemingly simple request could end up in a major political pain in the ass in your institution. If EH&S finds a problem like the above, then they can take cover in the fact that they were just doing their job. If you are responsible for finding it, you may not have that kind of protection. Rest assured, if a major problem ensues, EH&S will be more than happy to pass the credit to you.
On Thursday, May 17, 2001 5:27 PM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } } Hi: } } This is a weird question, it could only happen in Santa Cruz. } } One of our EH&S people stopped by today to ask if we could analyze some } dust from one of the science buildings on campus. I said 'Maybe' and } listened to the rest of the story. } } He told me that a new custodial employee was afraid that the dust in one of } the science buildings was full of toxic material and wanted it tested. The } building is a 30 year old structure that has had various sorts of biology } and chemistry teaching and research labs. As far as I know there has never } been any sort of problem there and EH&S keeps track of things like } radiation and highly toxic situations. } } I was reluctant to get involved. Our EDS could probably see some things and } I could recognize some mold spores and cotton fibers, but who am I? I don't } have any training or certification in toxic dust analysis. Plus I probably } already think this guy is a little paranoid about dust, so I probably } wouldn't find anything even if it was there. } } Are there labs that can do this sort of analysis and can certify that there } is nothing toxic to worry about in a sample of dust? How does one sample } for these things and how do we respond to this kind of request. What are } the odds that the dust is anything more than just dust? What is dust } anyway? } } I promised our EH&S guy that I would ask around among the smartest group of } folks I know. So what do you think? } } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Whoops in my last mailing my spell checker helped me to send an error in the last paragraph about printers. The 20 um should have been 200 dpi in the sixth line below:-
} Krzystof } you will probably get lots of answers to this but here goes: } {SNIP} } This whole situation is made worse, of course, by claimed resolutions of } printers. If it's an inkjet or laser printer then the claim of 1200 dpi } for output is nearer to 20um (or less depending on whose figures you } use) because dithering of several ink spots is required to produce } shades of grey or tones of colour whereas dye sublimation and image } printers usually achieve their actual figure claimed figures of 200 or } 300 dpi. So there's no point in creating an image with 400 dpi onto an } inkjet print because the maximum that could be represented by the } printer would be about 200 dpi.
I guess it just shows how easy it is to mix up pixel size and dpi.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
I would admit I am not an expert and collect a quick EDX spectrum for heavy metals and have a look for asbestos fibres.
Dave
On Thu, 17 May 2001 15:26:34 -0700 Jon Krupp {jmkrupp-at-cats.ucsc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } } This is a weird question, it could only happen in Santa Cruz. } } One of our EH&S people stopped by today to ask if we could analyze some } dust from one of the science buildings on campus. I said 'Maybe' and } listened to the rest of the story. } } He told me that a new custodial employee was afraid that the dust in one of } the science buildings was full of toxic material and wanted it tested. The } building is a 30 year old structure that has had various sorts of biology } and chemistry teaching and research labs. As far as I know there has never } been any sort of problem there and EH&S keeps track of things like } radiation and highly toxic situations. } } I was reluctant to get involved. Our EDS could probably see some things and } I could recognize some mold spores and cotton fibers, but who am I? I don't } have any training or certification in toxic dust analysis. Plus I probably } already think this guy is a little paranoid about dust, so I probably } wouldn't find anything even if it was there. } } Are there labs that can do this sort of analysis and can certify that there } is nothing toxic to worry about in a sample of dust? How does one sample } for these things and how do we respond to this kind of request. What are } the odds that the dust is anything more than just dust? What is dust } anyway? } } I promised our EH&S guy that I would ask around among the smartest group of } folks I know. So what do you think? } } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
On Thu, 17 May 2001 17:53:04 -0500 Jae Kim {jkim-at-bnl.gov} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear All: } } I would like to get an idea about the way to look at a morphology of a nano } sized powder sample in TEM. } } I need to take TEM morphology pictures from MgO powder (from nano meter to } a few micron size distribution) using a Cu grid. To spread a MgO particle } on the grid, I will spray some kinds of alcohol on the powder which resides } on the grid. Here, I would like to know what kinds of alcohol can be used } to get a clear image without melting the holey carbon film on the grid. } } I appreciate your comments. } } Jae } } ******************************** } Jae-yong Kim } Post-doctoral Research Associate } Chemistry Department } Brookhaven National Laboratory } Upton, NY 11973 } Tel:631-344-4317 } Fax:631-344-5815 } ******************************** }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I can't think of any electron dense spheres of the size that you suggest. The nearest I have seen is approx. 1um diameter latex spheres which are probably available from most suppliers. If you want electron density you could perhaps consider fixing some in a 1 or 2% solution of osmium tetroxide for 1 hour or more. You could then wash and examine them to see if they have enough density, retain size and shape. An alternative might be to gold sputter coat latex beads but they would have to be dried down then resuspended and the gold might wash off. I haven't tried either of these but if you use them and they work I would love to know.
Malcolm
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Nicol Aitken wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hello Listers, } } I was wondering if anyone out there can help me out. I am looking } } for a solution of uniform particles (spheres would be nice) of a heavy } } material so that it will show up nicely in backscatter. I also require } that } } the particle size be between 2 and 5 micron. If anyone knows of something } } like this and where I can order it from, It would help me out. } } Thanks in advance } } Nick
Although not accredited to do this kind of work "officially", I once put some dust collected from air filters in a building in the SEM. A friend of mine was the enivironment manager for a local communications company, and they were concerned about toxins in the workplace. The mold "Stachybotris" was one thing we were looking for (but didn't find). We also found no aluminosilicate fibres which could have been asbestos. We did find clay particles, combustion products, lots of little "organic" fibres (wool? polyester? cotton?) and assorted bits of micro-crud (that's a technical term). EDS analysis of some bits showed fair amounts of titanium, which I understand is a common ingredient in paints. So there wasn't anything obviously very worrisome in there. But never underestimate the power of Stachybotris - we had a large part of the Institute here shut down and then rebuilt a couple years ago because of it. Cost God knows how much, and people here are still concerned if someone finds a black stain on anything that could be damp. Anyway, dust analysis can be a pretty interesting exercise - I wish I had a copy of the McCrone Particle Atlas - it probably would have helped a lot.
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia Canada
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 17, 2001 7:26 PM
Listers -
Those of you with LaB6 experience - is it possible for one to fail in such a way that it still shows a "normal" emission current on all the gauges, yet is not producing a findable beam? Last Monday we stripped our ESEM down for a bi-annual cleaning, inspected everything, including the LaB6 (which looked fine, and had been working well up to that point), put the whole thing back together, and were unable to find a beam. Now we've done this any number of times in the last 8 years, and it's never more than a few minutes to find the beam afterwards. Turn the condensor down low, select a 3mm "hole" instead of a projection aperture, and bingo, there it is - a big, shiny, beam like a floodlight. This time, nothing. Not a glimmer of "light" on the monitor, even though the LaB6 is "on", with 15KeV, and a "normal" emission current. We've had the column back apart 4 times now, to see if there's some physical blockage somewhere which might prevent the beam from reaching the chamber, but everything looks great. Even checked the performance of the chamber isolation valve, in case it was failing to open, but it's working properly. So I'm left with thinking maybe it's the LaB6, but in the past when we had one fail, it either suddenly began producing a strange, assymetrical beam, with normal emission current readings, or no beam at all, with no emission current showing. What gives? We have a new LaB6 on order, as a spare, so when it arrives I'll put it in, unless somebody has any other ideas....
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
Did anyone mention that in the video world, "resolution" has long meant half of the value as used by computer people?
Television technicians (or composite video monitor specs, etc.) will still speak of "resolution" as in "lines of resolution" as in the number of vertical black lines that can be discerned apart from a white background.
Some time ago, someone in our chemistry department made a aqueous suspension of silica microspheres, about half a micron in diameter. If this was allowed to dry out, the spheres formed a close-packed lattice, similar to that in opal which gives rise to the lovely diffraction colours. When examining this lattice under the SEM, we found that this made an ideal target for the AUTO aSTIGmatism control, so when examining something bland such as a fine textured plastic surface, one could use a clump of these spheres as a target and then have the optics just right for examining the bland specimen.
Alas, the suspension of spheres has degenerated into some sort of "goo", and no more is available. Are there any other sources of spheres of this size? The 1 to 8 micron space spheres are probably a little too big, and I am a bit wary of what magnetite spheres might do in the electric field of the SEM.
+----------------------------------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------------------------------+
I wasn't involved in the NIST (NBS then - before my time) work but a quick search of NASA web brought up this report and many more on the microspheres: http://samson2.msfc.nasa.gov/fame/exps/van-011.html it contains references to journal articles for those interested in more information. SRM 1960 and 1961 are listed as available as Standard Reference Materials at http://srmcatalog.nist.gov/ Scott
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } SPI Supplies placed an order for these made-in-space microspheres (in the } late 1970's) when the program was first announced, and we had the } distinction of being the world's first purchaser of something actually made } in space! I am not sure that got us anything other than a feature one night } on the Nightly News. But despite all the publicity, at the time, that was } given out by NASA, I am unaware of anyone who ever published anything } showing that the "perfectly round" aspects of these microspheres (or any } other aspect for that matter) was anything above and beyond what is } routinely made on earth! We ourselves could not find that superiority, but } perhaps someone better that ourselves did succeed in finding such a } difference. } } The last I heard about that was that someone at Lehigh University had some } funding (remember we are talking about the early 1980's) to try to find that } these spheres did, in some way go beyond what could be made on earth, but I } never heard anything after that. } } BTW, we subdivided the vial purchased form NASA, and advertised them (as we } were encouraged to do) and over the years, as I met people at our exhibit } booth at trade shows, I never encountered anyone who found, when examining } these spheres, anything "above and beyond" earth made spheres. When the } set of samples was sold off, we never replenished our stock because we could } not in good conscience tell customers that the far higher price could be } justified in terms of there really being something special. }
------------------------------------------------------------------ Scott Wight fax: 301-417-1321 NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | email: scott.wight-at-nist.gov Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
I will just add my personal variations to the general method described by Geoff McAuliff:
Fjx & dehydrate (ethanols) as usual.
I have found that the following "hybrid" Epon analog separates cleanly from the culture dish just about every time. I Use LLX112 and DMP-3o, both from Ladd, and DDSA and NMA from Electron Microscopy Sciences. I have no clue why this particular mix works best, but a number of years ago, I got my hands on just about every component from every manufacturer and found that this mix did not react with the dishes at all, while many of the others did to some extent. I'd previously assumed that these components were all manufactured by one lab and just sold by the various vendors under their individual names, but the LX1l12 looks different than the other Epon812 substitutes.
The other thing that I do differently is in the actual embedding. I put a thin layer of the mixed resin into the dish (2 mm?), and insert tubes (made by cutting the pyramidal ends off of BEEM capsules)over areas of interest. I partially polymerize this (overnight) and in the morning, I fill JUST THE TUBES the rest of the way with more resin and then fully polymerize it all. When Its "cooked" I take a pair of needlle-nosed pliers, grab a tube and break it out with a slight twist to apply shearing force. It comes out cleanly. sometimes a small bit of the dish will come with it, but that pops right off when you start to trim the block. I just took a batch out of the oven this morning. I have nice clean faces with the cells clearly visible.
good luck, Lee Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I am posting this for Ricardo Drut. Ricardo wants to know if anyone has used Tissue -Tek Tissue-Clear as a substite for Xylol, and how do you like it. You may respond to Ricardo at his email address. Thanks, Teresa
} ¿Alguien usa este producto como aclarante sustituto del xilol? ¿Con qué } resultados?
You might try talking with Jim McCarthy. Jim is part of our EH&S group in our Sudbury Lab. Jim has had experience disposing of radioactive sources. Jim's contact information follows.
Lenn C. Kupferberg Principal Physicist Raytheon Company Electronic Systems Lexington Laboratory 131 Spring Street Lexington, MA 02421-7803 (781)860-3082
James J. McCarthy Raytheon Company C3I Sudbury Laboratory 528 Boston Post Road Sudbury, Mass. 01776 (978)440-4047 James_J_McCarthy-at-raytheon.com
---------------------- Forwarded by Lenn C Kupferberg/RES/Raytheon/US on 05/18/2001 08:09 AM ---------------------------
Norman C Miller 05/17/2001 06:02 PM
To: Lenn C Kupferberg/RES/Raytheon/US-at-MAIL cc:
We have an Americium x-ray source that we no longer need. My understanding is that disposal of this is extremely expensive. Has anyone dealt with this problem and can give me some suggestions? Thank you, Lynne C. Garone Polaroid Corp. 1265 Main St. W4-1D Waltham, MA 02451-1714 (781) 386-1446 GaroneL-at-Polaroid.com
We have a Philips 400T for sell. It had been kept on contract. And the whole system, including EDS and STEM, has been in full function before it was moved out of the lab and packed to have space for a 200kV scope. Anybody interested in purchasing please contact me off line. Thanks
Chaoying Ni Director of Electron Microscopy Lab Dept. of Materials sci. and Eng. University of Delaware Newark, DE 19716 cni-at-udel.edu
I'm going to build an EM lab server to run web and other applications, to share files, especially to have an interactive wed scheduling software to schedule and monitor usage of our 2 TEMs and 2 SEMs. Right now I have narrow down the platform to Linux and Windows 2000. Could you please give me some inputs concerning the pros and cons of these two operating systems? Any comments and suggestions on the hardware selection are also more than welcome. Thanks in advance!
Chaoying Ni Electron Microscopy Lab Dept. of Materials sci. and Eng. University of Delaware Newark, DE 19716 cni-at-udel.edu
} } This is a weird question, it could only happen in Santa Cruz. } } One of our EH&S people stopped by today to ask if we could analyze some } dust from one of the science buildings on campus. I said 'Maybe' and } listened to the rest of the story. }
I have several times been asked by our Environmental Medical people to analyze specific samples of deposits found in various places around the campus, usually finding them to be quite benign (steel filings, plaster dust, (presumably from sheetrock), soap deposits, etc. etc). The steel and plaster I did with the SEM, other things (like the soap) I passed to other members of our team here for techniques like FTIR.
Taking a generalized "dust" sample and proving beyond doubt that it is harmless is a totally different issue.
Dust, of course, is simply whatever happens to fall on the floor that is too small to be picked up piece-by-piece. Dust in the bedroom at home is very different from dust in a laboratory. In some cases it will be dominated by biological material (human skin and hair, and fibers from clothing). In other cases by residue from construction activities in other parts of a building. Depending on the ventilation arrangements, particulate matter from outside may be brought in. This could contain soots from internal combustion engines or oil or coal fired electric plants, or even wildfires, or fine mineral particles (sometimes from very remote sources), as well, possibly, as aerosols or other particulates from local industrial activity. There will always be material brought in on shoes, especially in the winter, and, of course, wear debris from traffic in the floor.
Can a non-specialist determine whether it is "hazardous"? (What is "hazardous", anyway?) Not in my book. There is just too much to look for, especially if you are trying to reassure a paranoid and suspicious person. One example - does finding Si and Al in a particle tell you that it is asbestos? Of course not. Does a failure to find Si and Al mean asbestos is absent? Again, of course not - it could simply be a minor constituent, and the elemnts below the detectability limit.
Good luck!
Tony.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
I have done bunches of SEM shots of house dust. Really quite interesting. This was not a search for a smoking gun, just an interesting exercise. I found (all very small) fibers, skin flakes, odds and ends of all other sorts of things. There we no mites or other live (before SEM) or dead organisms that I could find. The specimens were taken on a sticky stub sitting on top of a TV so the static electricity would draw room junk to the stub. It did.
I have several other ones at floor level which I have not yet imaged. I usually wait about 4 months before coating and imaging. This guarantees a good smattering of crud.
Old buildings would have, I think, Aspergillis mold. This would explain respiratory problems and many ills. There are other mold types of course.
gary g.
At 03:26 PM 5/17/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in studying mitochondrial ultrastructure in various tissues during hypoperfusion and was wondering if there were any specific TEM stains/techniques or immunofluorescent techniques for examining the mitochondrial permeability transition? Thanks.
Michael J. Menconi, Ph.D.
Department of Surgery Brigham and Women's Hospital Medical Research Building, Room 502 75 Francis Street Boston, MA 02115 Phone: 617-278-0693 FAX: 617-582-6047 Email: mmenconi-at-partners.org
Nick - Check out Duke Scientific (dukescientific.com) who sell a range of different size microspheres for calibration purposes. Many are either glass, silica or polystyrene which won't specifically meet your high Z requirement, however, you could easily coat them with Au or Pd or some other metal.
Dave Joswiak Dept. of Astronomy University of Washington Seattle, WA 98195 joswiak-at-astro.washington.edu (206)543-7702
On Thu, 17 May 2001, Nicol Aitken wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } Hello Listers, } } I was wondering if anyone out there can help me out. I am looking } } for a solution of uniform particles (spheres would be nice) of a heavy } } material so that it will show up nicely in backscatter. I also require } that } } the particle size be between 2 and 5 micron. If anyone knows of something } } like this and where I can order it from, It would help me out. } } Thanks in advance } } Nick } }
Prof. Barajon, take a look in our paper entitled "SANTOS & MARIATH, 1997. A simple method for fixing, dehydrating and embedding pollen tubes cultivated in vitro for optical and transmission electron microscopy. Biotechnic & Histochemistry, 72(6): 315-319" . Could be your problem solution! Hoping success! Mariath
----- Original Message ----- } From: "Geoff McAuliffe" {mcauliff-at-umdnj.edu} To: "Prof. Barajon" {isabella.barajon-at-unimi.it} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 17, 2001 4:31 PM
Dear Paul, I seem to remember using a 3% solution of methyl cellulose to slow down paramecium.I suppose the concentration can be adusted for larger beasties.
Christine Richardson E.M.Unit University of Durham Dept.Biological Science
Durham England -----Original Message----- } From: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com [mailto:"Pbgrover-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, May 15, 2001 9:46 PM To: microscopy-at-sparc5.microscopy.com
Dear Listers,
I have some 5-to-8 year-olds who really enjoy the microworld, but whose hands can't keep up with speedy microbeasties under the lens. I remember from years past that there was a product (I think it was called 'Proto-Slo', or similar) that was viscous, roughly iso-osmotic, and designed to slow the critters down for easier observation. Is it still available? If not, does anyone have a recipe for a home-brewed equivalent?
Thank you, thank you, thank you,
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
I am trying to find out whether there is anyone or organization that is available to service a Peak Instrument Co. Focus MCS 4 Multi Element Crystal Spectrometer Model O,C,N,B integrated with a PGT Imix System.
Thanks for your help.
Stanley H. Gelles Senior Group Leader CC Technologies 614-761-1214 FAX 614-761-1633
Dear Paul, I seem to remember using a 3% solution of methyl cellulose to slow down paramecium.I suppose the concentration can be adusted for larger beasties.
Christine Richardson E.M.Unit University of Durham Dept.Biological Science Durham England
Dear Paul, I seem to remember using a 3% solution of methyl cellulose to slow down paramecium.I suppose the concentration can be adusted for larger beasties.
Christine Richardson E.M.Unit University of Durham Dept.Biological Science
Durham England -----Original Message----- } From: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com [mailto:"Pbgrover-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, May 15, 2001 9:46 PM To: microscopy-at-sparc5.microscopy.com
Dear Listers,
I have some 5-to-8 year-olds who really enjoy the microworld, but whose hands can't keep up with speedy microbeasties under the lens. I remember from years past that there was a product (I think it was called 'Proto-Slo', or similar) that was viscous, roughly iso-osmotic, and designed to slow the critters down for easier observation. Is it still available? If not, does anyone have a recipe for a home-brewed equivalent?
Thank you, thank you, thank you,
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
Bad luck indeed. It would be helpful to know what make of LaB6 you are using because the physical construction may have a great deal to do with the diagnosis of your problem. Anyway, what I suspect, given the eight years of experience with your instrument is that nothing incorrect was done on your part but rather, the LaB6 itself has gone bad. If you have not done so yet, it might be helpful to recenter the tip and while you have everything out, take a very close look at the crystal itself. They have been known to seperate from their heater block and thermal shock can cause the crystal to crack, causing mechanical displacement. The crystal may have actually fallen off the block and all you are doing is heating the carbon heater. If you can, it would eliminate a bunch of variable if you would install a plain old hair-pin Tungsten Filament and see how the instrument behaves.
Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone 512/282-5507 FAX 512/280-0702
Sustaining Member - MICROSCOPY SOCIETY OF AMERICA ----- Original Message ----- } From: "Frank Thomas" {thomasf-at-AGC.BIO.NS.CA} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 18, 2001 7:53 AM
If your column has a column liner and an aperture at the bottom of the liner, could it have gotten some crud on it during your work on the system? I would think that with this aperture blocked, there would not be any emission current.
What about carefully checking the SE detector? Faraday shield (make sure it is in-place correctly), check the scintillator disc, etc. It sounds like the beam is there but the SEs are not being detected. If you have a Faraday cup in or for your specimen holder, use a large aperture (or none) and see if you get specimen current (I use the stage alarm BNC connection to do this with a 10Meg Ohm DVM. I=V/10^6).
Just a thought.
gary g.
At 05:53 AM 5/18/2001, you wrote:
} Listers - } } Those of you with LaB6 experience - is it possible for one to fail in such a } way that it still shows a "normal" emission current on all the gauges, yet } is not producing a findable beam? } Last Monday we stripped our ESEM down for a bi-annual cleaning, inspected } everything, including the LaB6 (which looked fine, and had been working well } up to that point), put the whole thing back together, and were unable to } find a beam. Now we've done this any number of times in the last 8 years, } and it's never more than a few minutes to find the beam afterwards. Turn the } condensor down low, select a 3mm "hole" instead of a projection aperture, } and bingo, there it is - a big, shiny, beam like a floodlight. } This time, nothing. Not a glimmer of "light" on the monitor, even though the } LaB6 is "on", with 15KeV, and a "normal" emission current. We've had the } column back apart 4 times now, to see if there's some physical blockage } somewhere which might prevent the beam from reaching the chamber, but } everything looks great. Even checked the performance of the chamber } isolation valve, in case it was failing to open, but it's working properly. } So I'm left with thinking maybe it's the LaB6, but in the past when we had } one fail, it either suddenly began producing a strange, assymetrical beam, } with normal emission current readings, or no beam at all, with no emission } current showing. What gives? We have a new LaB6 on order, as a spare, so } when it arrives I'll put it in, unless somebody has any other ideas.... } } Frank Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada
In response to Ron Vane's response to the dust analysis question: I think you, Ron, should keep your political views about animal testing out of this forum, especially since they seem to be oversimplified.
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 jlagoy-at-bodycote-imt.com
the stachybotris link http://gcrc.cwru.edu/stachy/
At 12:35 AM 5/18/01 -0500, Allen R. Sampson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
this is the airborne fungi database http://www.bio.psu.edu/People/Faculty/Whittam/apdbase/fungus.html
At 12:35 AM 5/18/01 -0500, Allen R. Sampson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form. It was submitted by (colonel-at-natca.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, May 18, 2001 at 17:55:26 ---------------------------------------------------------------------------
Email: colonel-at-natca.net Name: M. B. Ingersoll
Organization: Schufeldt Academy
Education: 6-8th Grade Middle School
Location: Euless, TX, USA
Question: We are a homeschooling family. I have been all over the Internet trying to find information (and hopefully a manual) for a recently purchased, refurbished microscope.
The unit is a monocular, three objective (10x, 40x, 100x) self illuminated model with mechanical staging and a variable condensor. I am an Air Traffic Controller and not a microscopist so please forgive me if I misuse any terminology.
From the markings I believe the manufacturer was SPI or PSI (the letters S-P-I are arranged in that order in a diamond shaped logo). There are two more numbers under the logo; 1804 (a model number?) and 23490 (preceded by "No." - presumably a serial number?). I was told the unit is aproximately 30 years old.
Lastly, there is a small paper label on the underside of the base with the word "JAPAN" on it.
Can you provide any information or point me to another source?
Ultimately this is a matter of personal preference and other requirements. Are you an experienced Linux or Unix system administrator? Will you be responsible for administration and security? What would your local network administrator prefer/suggest? What languages and development environments for you applications do you want to use? What database system will you use? These are just some examples of the kinds of questions you should address first.
We have a Windows 2000 Server, with IIS 5.0 and FrontPage Server Extensions and MS Access for databases. We chose this system for several reasons: myself and the other developer are most comfortable developing software using Visual Basic and vbscript; it is much easier to build and administrate than a Linux system (at least for me), although I believe in any case the time investment would be lower; at an educational institution cost of the licenses is not much of an issue; you can automatically receive update notifications (security); we just recently added a RAID 1 (mirroring) system to protect from data loss and down time due to hard drive failures (trivial to set up); preferred OS of our local network administrator; among others.
Of course, you might answer these questions entirely differently and your answer might be Linux with Apache, java server pages(JSP) or PHP, MySQL, etc.
Chao-Ying Ni wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear colleagues, } } I'm going to build an EM lab server to run web and other applications, } to share files, especially to have an interactive wed scheduling software } to schedule and monitor usage of our 2 TEMs and 2 SEMs. Right now I have } narrow down the platform to Linux and Windows 2000. Could you please give } me some inputs concerning the pros and cons of these two operating } systems? Any comments and suggestions on the hardware selection are also } more than welcome. Thanks in advance! } } Chaoying Ni } Electron Microscopy Lab } Dept. of Materials sci. and Eng. } University of Delaware } Newark, DE 19716 } cni-at-udel.edu
Respectfully, views on animal research seem to be perfectly suited to civil discourse on this venue. Probably a significant number of the listserver are involved with animal research. Issues affecting our colleagues are definitely worth discussion and not to be censored by labeling those opinions as political and oversimplified.
Chuck Butterick Engineered Carbons, Inc. Borger, TX 79008
____________________________
In response to Ron Vane's response to the dust analysis question: I think you, Ron, should keep your political views about animal testing out of this forum, especially since they seem to be oversimplified.
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 jlagoy-at-bodycote-imt.com
Go with the one you have the in house expertise to support. If you have to support it yourself and don't want to spend time learning Linux or Unix Windows is the easiest way. If you want to add a skill to your resume Linux or Unix is a very marketable skill and makes a much more stable and secure platform.
My Linux server has been up 252 days and it has two and a half years since any software updates have been done except to patch some security flaws. The only problems I have had were a break in and a processor that went bad. I have had zero problems from software instability. But I had a good system administrator set it up.
The secret to a stable system is once it is running like you want it leave it alone. If you want to tinker with something get another system on line to play with don't do it to the one that is in production. Once it is working on the other system put it on the production system.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} } Dear colleagues, } } } } I'm going to build an EM lab server to run web and other applications, } } to share files, especially to have an interactive wed scheduling software } } to schedule and monitor usage of our 2 TEMs and 2 SEMs. Right now I have } } narrow down the platform to Linux and Windows 2000. Could you please give } } me some inputs concerning the pros and cons of these two operating } } systems? Any comments and suggestions on the hardware selection are also } } more than welcome. Thanks in advance! } } } } Chaoying Ni } } Electron Microscopy Lab } } Dept. of Materials sci. and Eng. } } University of Delaware } } Newark, DE 19716 } } cni-at-udel.edu } }
SPI (and I forget what this stands for) is the company that you want. They may still be in business and you might check through the search engines for them one thing that I remember about SPI is that they also imported micrometers and other materials also related to the precision machine industry.
We regret that we have none of this company's catalogs or info here in the archive, our collection of catalogs and inst. books is growing though, If something comes in we will be happy to let you know. However if you do obtain any documentation on this and wish to forward us a Xerox please feel free to send to:
coury house / smecc 5902 w palmaire ave glendale az 85301
Good luck in the hunt!
Ed Sharpe Archivist for SMECC
} Subj: Ask-A-Microscopist:Looking for a manual/info on a LM } Date: 5/18/01 7:33:30 PM US Mountain Standard Time } From: colonel-at-natca.net } To: Microscopy-at-sparc5.microscopy.com } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form. It was submitted by } (colonel-at-natca.net) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, May } 18, 2001 at 17:55:26 } --------------------------------------------------------------------------- } } Email: colonel-at-natca.net } Name: M. B. Ingersoll } } Organization: Schufeldt Academy } } Education: 6-8th Grade Middle School } } Location: Euless, TX, USA } } Question: We are a homeschooling family. I have been all over the } Internet trying to find information (and hopefully a manual) for a } recently purchased, refurbished microscope. } } The unit is a monocular, three objective (10x, 40x, 100x) self } illuminated model with mechanical staging and a variable condensor. } I am an Air Traffic Controller and not a microscopist so please } forgive me if I misuse any terminology. } } From the markings I believe the manufacturer was SPI or PSI (the } letters S-P-I are arranged in that order in a diamond shaped logo). } There are two more numbers under the logo; 1804 (a model number?) and } 23490 (preceded by "No." - presumably a serial number?). I was told } the unit is aproximately 30 years old. } } Lastly, there is a small paper label on the underside of the base } with the word "JAPAN" on it. } } Can you provide any information or point me to another source? } } Thank you! } } --------------------------------------------------------------------------- } }
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} SPI (and I forget what this stands for) is the company that you want. {BR} They may still be in business and you might check through the search {BR} engines for them one thing that I remember about SPI is that they also {BR} imported micrometers and other materials also related to the precision {BR} machine industry. {BR} {BR} We regret that we have none of this company's catalogs or info here in the {BR} archive, {BR} our collection of catalogs and inst. books is growing though, If something {BR} comes in {BR} we will be happy to let you know. However if you do obtain any documentation {BR} on this and wish to forward us a Xerox please feel free to send to: {BR} {BR} coury house / smecc {BR} 5902 w palmaire ave {BR} glendale az 85301 {BR} {BR} Good luck in the hunt! {BR} {BR} Ed Sharpe Archivist for SMECC {BR} {BR} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} Subj: {B} Ask-A-Microscopist:Looking for a manual/info on a LM {/B} {BR} Date: 5/18/01 7:33:30 PM US Mountain Standard Time {BR} {I} From: colonel-at-natca.net {BR} To: Microscopy-at-sparc5.microscopy.com {BR} {/I} {BR} {BR} {BR} {BR} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} -----------------------------------------------------------------------. {BR} {BR} {BR} Below is the result of your feedback form. It was submitted by {BR} (colonel-at-natca.net) from {BR} http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, May {BR} 18, 2001 at 17:55:26 {BR} --------------------------------------------------------------------------- {BR} {BR} Email: colonel-at-natca.net {BR} Name: M. B. Ingersoll {BR} {BR} Organization: Schufeldt Academy {BR} {BR} Education: 6-8th Grade Middle School {BR} {BR} Location: Euless, TX, USA {BR} {BR} Question: We are a homeschooling family. I have been all over the {BR} Internet trying to find information (and hopefully a manual) for a {BR} recently purchased, refurbished microscope. {BR} {BR} The unit is a monocular, three objective (10x, 40x, 100x) self {BR} illuminated model with mechanical staging and a variable condensor. {BR} I am an Air Traffic Controller and not a microscopist so please {BR} forgive me if I misuse any terminology. {BR} {BR} From the markings I believe the manufacturer was SPI or PSI (the {BR} letters S-P-I are arranged in that order in a diamond shaped logo). {BR} There are two more numbers under the logo; 1804 (a model number?) and {BR} 23490 (preceded by "No." - presumably a serial number?). I was told {BR} the unit is aproximately 30 years old. {BR} {BR} Lastly, there is a small paper label on the underside of the base {BR} with the word "JAPAN" on it. {BR} {BR} Can you provide any information or point me to another source? {BR} {BR} Thank you! {BR} {BR} --------------------------------------------------------------------------- {BR} {BR} {/BLOCKQUOTE} {BR} {BR} {BR} {/FONT} {/HTML}
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from rly-xd02.mx.aol.com (rly-xd02.mail.aol.com [172.20.105.167]) by air-xd02.mail.aol.com (v77_r1.36) with ESMTP; Fri, 18 May 2001 22:33:30 -0400 Received: from sparc5.microscopy.com ([206.69.208.10]) by rly-xd02.mx.aol.com (v77_r1.36) with ESMTP; Fri, 18 May 2001 22:33:03 2000 Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id SAA14967 for dist-Microscopy; Fri, 18 May 2001 18:27:20 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id SAA14964 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 18 May 2001 18:26:49 -0500 (CDT) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id SAA14957 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 18 May 2001 18:26:38 -0500 (CDT) Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {p05001900b72b6000cf44-at-[206.69.208.21]}
Below is the result of your feedback form. It was submitted by (colonel-at-natca.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, May 18, 2001 at 17:55:26 ---------------------------------------------------------------------------
Email: colonel-at-natca.net Name: M. B. Ingersoll
Organization: Schufeldt Academy
Education: 6-8th Grade Middle School
Location: Euless, TX, USA
Question: We are a homeschooling family. I have been all over the Internet trying to find information (and hopefully a manual) for a recently purchased, refurbished microscope.
The unit is a monocular, three objective (10x, 40x, 100x) self illuminated model with mechanical staging and a variable condensor. I am an Air Traffic Controller and not a microscopist so please forgive me if I misuse any terminology.
From the markings I believe the manufacturer was SPI or PSI (the letters S-P-I are arranged in that order in a diamond shaped logo). There are two more numbers under the logo; 1804 (a model number?) and 23490 (preceded by "No." - presumably a serial number?). I was told the unit is aproximately 30 years old.
Lastly, there is a small paper label on the underside of the base with the word "JAPAN" on it.
Can you provide any information or point me to another source?
You may have answered your own question. Opal makes a good SEM sample when fractured, acid etched and coated. A nicely packed lattice of very round spheres, I use one as simple test sample, along with a semiconductor die and NIST magnification and resolution standards. A small chunk can virtually last forever.
On Friday, May 18, 2001 7:57 AM, Robert H. Olley [SMTP:r.h.olley-at-reading.ac.uk] wrote: } } } Hello all Listers, } } Some time ago, someone in our chemistry department made a aqueous } suspension of silica microspheres, about half a micron in diameter. If } this was allowed to dry out, the spheres formed a close-packed lattice, } similar to that in opal which gives rise to the lovely diffraction } colours. When examining this lattice under the SEM, we found that this } made an ideal target for the AUTO aSTIGmatism control, so when examining } something bland such as a fine textured plastic surface, one could use a } clump of these spheres as a target and then have the optics just right } for examining the bland specimen. } } Alas, the suspension of spheres has degenerated into some sort of "goo", } and no more is available. Are there any other sources of spheres of } this size? The 1 to 8 micron space spheres are probably a little too } big, and I am a bit wary of what magnetite spheres might do in the } electric field of the SEM. } } +----------------------------------------------------------------+ } Robert H.Olley } J.J.Thomson Physical Laboratory } University of Reading } Whiteknights } Reading RG6 6AF } England } +----------------------------------------------------------------+ } Phone: } {direct line +44 (0) 118 9318572 } {University internal extension 7867 } Fax: +44 (0) 118 9750203 } Email: R.H.Olley-at-reading.ac.uk } URL: http://www.reading.ac.uk/~spsolley } +----------------------------------------------------------------+ } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Frank, I thing you are very lucky to normally find the beam so quickly. We have an E3 ESEM and on that beast it can take ages to find the beam after routine service. Regards JVN
Frank Thomas wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers - } } Those of you with LaB6 experience - is it possible for one to fail in such a } way that it still shows a "normal" emission current on all the gauges, yet } is not producing a findable beam? } Last Monday we stripped our ESEM down for a bi-annual cleaning, inspected } everything, including the LaB6 (which looked fine, and had been working well } up to that point), put the whole thing back together, and were unable to } find a beam. Now we've done this any number of times in the last 8 years, } and it's never more than a few minutes to find the beam afterwards. Turn the } condensor down low, select a 3mm "hole" instead of a projection aperture, } and bingo, there it is - a big, shiny, beam like a floodlight. } This time, nothing. Not a glimmer of "light" on the monitor, even though the } LaB6 is "on", with 15KeV, and a "normal" emission current. We've had the } column back apart 4 times now, to see if there's some physical blockage } somewhere which might prevent the beam from reaching the chamber, but } everything looks great. Even checked the performance of the chamber } isolation valve, in case it was failing to open, but it's working properly. } So I'm left with thinking maybe it's the LaB6, but in the past when we had } one fail, it either suddenly began producing a strange, assymetrical beam, } with normal emission current readings, or no beam at all, with no emission } current showing. What gives? We have a new LaB6 on order, as a spare, so } when it arrives I'll put it in, unless somebody has any other ideas.... } } Frank Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
Below is the result of your feedback form. It was submitted by (hines-ta-at-actx.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 20, 2001 at 16:03:16 ---------------------------------------------------------------------------
Email: hines-ta-at-actx.edu Name: Tracey Hines
Organization: Amarillo College
Education: Undergraduate College
Location: Amarillo, Texas
Question: I am having a difficult time finding any biographical information on George Nomarski. Do you know of any web sites or books that will be useful in finding information like birthdate, etc.?
Someone gave me a recept to make a sample with Tin balls. You need a polished aluminium sample, a old vaccum coater with a unregulated low voltage/ high current supply (one with a big variac and a 4V-12V/1000A transformer). You put some tin (Sn) in a Mo or W boat, (and of course your sample in the oposite) and you heat it under vacuum very quickly. It makes an "explosion" coating, and you will find tin balls, with a great dispersion in size on your aluminium sample (a few 100 nm to a few microns). It is a good test for backskatered detector and for SE resolution. With some practice, you schould be able to control the size, size distribution and density of tin balls.
I didn't try this receipt (not enouhgt time to try all what I want ! ), but according to my experience in thin film technology, it seems to be credible.
I saw also something like that to sell in a microscopy supplies catalogue, but I don't remember where, I think it was someone in Germany.
By
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } Hello Listers, } } I was wondering if anyone out there can help me out. I am looking } } for a solution of uniform particles (spheres would be nice) of a heavy } } material so that it will show up nicely in backscatter. I also require } that } } the particle size be between 2 and 5 micron. If anyone knows of something } } like this and where I can order it from, It would help me out. } } Thanks in advance } } Nick } }
Below is the result of your feedback form. It was submitted by (meehall-at-mac.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 20, 2001 at 23:49:07 ---------------------------------------------------------------------------
Email: meehall-at-mac.com Name: michael n maloney
Organization: san francisco police dept
Education: Graduate College
Location: san francisco
Question: i'm looking for the most appropriate microscope for fine art photographs of diatom and radiolarian sized objects. i understand that a true darkfield illumination source is preferable. a support for a heavy camera is also required. I use 35 mm (Nikon F5) and a 4x5 camera. please give me some thoughts on this
I use both Windows 2000 and Linux and like them both but they are as different as night-and-day. When all is said and done, both can be reliable, workhorse servers.
Here are a couple of issues you might consider: Administration - Windows 2000 Server is not easy to manage but it is substancially easier than Linux. Most Linux servers are set up by long-time Unix gurus. To them the obscure commands and syntax are second nature and they understand the sublties of network administration. If this doesn't describe you then it is probable that the lack of handholding in the Linux world will make configuring your server an adventure. Some of the standard Linux installs (Red Hat for example) make setting up a server easier but at the expense of leave gapping security holes. Clients - Are your client computers running Unix, Windows or ? While Linux can interoperate with Windows, it is easier to configure a Linux system to work with other Linux/Unix systems. Similarly, Win2k is easy to configure to work with Windows and a little harder to configure to work with Unix/Linux.
In summary, either one can work. Linux requires more knowledge and experience but the price is right.
************************************************************* Postdoctoral Position in Transmission Electron Microscopy - Rice University
Rice University, Department of Mechanical Engineering and Materials Science, located in Houston, TX, is seeking candidates for a postdoctoral position in transmission electron microscopy of multicomponent oxide thin films. Applicants should have extensive and demonstrated experience in several areas of TEM and a strong background and interest in materials problem solving. Preference will be given to candidates with experience in several TEM techniques. Facilities at Rice include a JEOL 2010, field-emission SEM and high-resolution X-ray diffractometers, as well as state-of-the-art sample preparation. The project will be carried out in close collaboration with the University of Houston using a state-of-the-art field-emission TEM (JEOL 2010F), with annular dark-field detector, Oxford link EDS, Gatan GIF and STEM capabilities. The position is available immediately. Duration about 1-2 years, salary is commensurate with qualifications. Candidates with a Ph.D. in Materials Science or Physics will be given preferred consideration. Interested candidates should send a curriculum vitae, publication list and the names of at least three references with their contact addresses to:
Prof. Susanne Stemmer Rice University Department of Mechanical Engineering and Materials Science MS 321 6100 Main Street Houston, TX 77005-1892 stemmer-at-rice.edu *************************************************************
We microscopists have focused tools with narrow fields of view. Toxicity is a very broad problem which our tool can see only a small portion.
My reply was not meant to reflect on my views of animal research but was an example of the problem of satisfying two very different portions of the technophobic community around us. As such it was very oversimplified.
I agree that animal research is not a good topic for this forum.
Jon: Let's try to get back to a reasonable answer here. Like you, I think that getting into it yourself can be problematic. So, for labs that do the work: McCrone and RJLee are the two I am most familiar with. I would guess there are others, and have no interest in the two mentioned, nor do I have anything against any lab. Labs like this are easily searchable on the internet, and I think you could also find them under one of the microscopy web resources (sorry-having a total medicare moment on names and such). Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals On Thu, 17 May 2001 15:26:34 -0700, Jon Krupp wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi: | | This is a weird question, it could only happen in Santa Cruz. | | One of our EH&S people stopped by today to ask if we could analyze some | dust from one of the science buildings on campus. I said 'Maybe' and | listened to the rest of the story. | | He told me that a new custodial employee was afraid that the dust in one of | the science buildings was full of toxic material and wanted it tested. The | building is a 30 year old structure that has had various sorts of biology | and chemistry teaching and research labs. As far as I know there has never | been any sort of problem there and EH&S keeps track of things like | radiation and highly toxic situations. | | I was reluctant to get involved. Our EDS could probably see some things and | I could recognize some mold spores and cotton fibers, but who am I? I don't | have any training or certification in toxic dust analysis. Plus I probably | already think this guy is a little paranoid about dust, so I probably | wouldn't find anything even if it was there. | | Are there labs that can do this sort of analysis and can certify that there | is nothing toxic to worry about in a sample of dust? How does one sample | for these things and how do we respond to this kind of request. What are | the odds that the dust is anything more than just dust? What is dust | anyway? | | I promised our EH&S guy that I would ask around among the smartest group of | folks I know. So what do you think? | | | Jonathan Krupp | Microscopy & Imaging Lab | University of California | Santa Cruz, CA 95064 | (831) 459-2477 | jmkrupp-at-cats.ucsc.edu | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Prof Barajon: Just a minor addition to the other comments (I have used variations of all of them). While not familiar with the referenced paper, I would add that you can (perhaps should) reduce your fixation times. I use 45 to 60 min in the aldehyde and 30 to 45 min in osmium. I can't remember the references, but there is an older reference to using methylene blue for en bloc staining of the epoxy-embedded cells. This really enhances the ability to locate areas of interest after embedding. (I believe the paper was in Stain Technology, probably in the 70's). This must be a senior moment day--I can't remember details on anything.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals On Thu, 17 May 2001 10:44:03 +0200, Prof. Barajon wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Subject: TEM- cell colture fixation and embedding | | I need to study the ultrastructural aspects of a cell line grown on petri | dishes. Any suggestion for the best fixation and embedding protocol (no | pellets, just fixation and embedding in the dish itself) ? | | thanks | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I'm looking for a suggestion for appropriate film support grids for making EELS and EDS measurements of 1 to 10 nm particles. I would use plain carbon, except that the particles will be a mixture of carbide, oxide, metal and silicate phases. I need to measure the C, O, N, Si, Mg and Fe content of the particles, so C, SiO2 or Si3N4 support films all would make my life difficult.
Has anyone made Be film grids? Any other suggestions?
Thank in advance.
Rhonda Stroud
Research Physicist Naval Research Laboratory 4555 Overlook. Ave SW Washington, DC 20375
I'm looking for a second-hand 2000FX double tile holder, or the compatible 2-tile JEOL100 holder. Anybody having the information on this please let know. Thanks much!
Chaoying Ni EM Lab Materials Sci and Eng University of Delaware
I would agree that familiarity with the operating environment is important for timely performance of server administration tasks-documentation and advocacy groups exist for both platforms, however. It might be worth while to become familiar with a UNIX like operating system (such as Linux) if one has the luxury of time (I'll explain momentarily). Cost may be an issue-Linux is free. Both OSes run on the x86 architecture. It seems that your choice(s) of application software should be a heavywieght factor in your decision. At the risk of over-simplification I'll just say that most proprietary "out of the box" solutions are developed for Windows/NT. There is a huge installed base of Wintel machines, and there are powerful software development tools for this platform which allow software developers to develop large complicated programs very rapidly. Linux has the advantage that many programs written to run under the various flavors of UNIX will compile and run. Many cutting edge scientific applications, including image processing utilities, are developed by researchers who are experienced coders and prefer to work under the UNIX platform. The reason for this is partially historical-in the past only large UNIX mainframe servers had the horsepower for image manipulation. More often than not these software tools are available free of charge to fellow researchers and are provided as open-source (one can view and modify the original code to suit specialized situations). So it really boils down to an ease of use vs. scalability/extensibility issue with cost, and perhaps minor hardware decisions (sometimes peripherals are difficult to configure under Linux ie. video cards, scanners etc...). In the long run, however, time spent now learning to work with Linux may pay off: the heart of the OS (the kernel) evolves continually in response to the needs of users, and the kernel can be upgraded without disrupting other parameters of the OS. Also, Linux runs on most (if not by now all)commonly encountered chip architectures so one may have several different kinds of machine (PC, Mac, SUN) all running the same operating system.
Karl Garsha www.uwm.edu/~keg ----- Original Message ----- } From: "Chao-Ying Ni" {cni-at-udel.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 18, 2001 9:36 AM
Hi folks, We have an ancient ISI (Akashi) Mini-SEM which is need of a service/overhaul. Could anyone recommend service companies that would be able to deal with such an old instrument? Companies located in the southwest would be preferable. Thank you for your time, Nick Bulloss
****************************************** Nick Bulloss Analytical Facilities Technician Department of Geological Sciences University of Texas at El Paso 500 West University Ave El Paso, TX 79968-0555 Office: (915) 747-5440 Lab: (915) 747-5184 Fax: (915) 747-5073
Jon, Your question raises questions whose import we should all understand. Getting involved, as Dr. Moretz said is 'problematic', but my response is much stronger. Getting involved is potentially hazardous to your health! You have answered your big question. "Who am I?" And you mean, not merely how much of what is required can "I" accomplish? You are correct to hesitate. The implications of involvement in such an institutional question are NOT trivial. First of all, as an institutional employee, you will have NO legal credibility except where you assert a positive finding of institutional fault, and the institution, would then be obligated to find an external source to either defend or deny your assertion. The only way by which this morass can be avoided is if an institutional policy directs you to perform such internal testing in order to determine 'an indication'. Second, there is NO good ground for you in such a question, and the person who requested some help was ill-advised and, worse, ill-prepared, for this personnel problem. The question raised by the 'concerned' employee was a "black hole". His/her supervisor should have immediately sent her/him to the appropriate Human Resource representative. The employee's concern was instant grounds for transfer, and immediate grounds for the institution to guard itself from her/his problem. Human resources should have a record of personnel performance in the workplace in question. If there has been an attendance problem due to illness among individuals working in the building or in spaces within the building, Human Resources should probably initiate an investigation or an upward 'chain of concern' if there is no clear policy that they can follow. You have raised the question of our own awareness and understanding of the difference between good science and good evidence as viewed by different interests within our society. We, who have been trained through graduate school for the jobs we have performed PRESUMABLY understand the inherent hazards of our jobs. Oil vacuum systems in cool but poorly ventilated rooms. Photographic darkrooms with inadequate ventilation. Formaldehyde, formalin, paraformaldehyde, paraldehyde, all peroxides and peroxide formers, X-Rays, 32P, Xylene, Propylene Oxide, MeOH, dissolving osmium tetroxide crystals, Toluene, human tissues and fluids, Latex, Hepatitis B, etc. Most of us as microscopist have taken 'the list' with the job, and very few of us have been crippled by the hazards. Yet; hazards there are. Those otherwise capable individuals who have been selected out of our zone in the sciences have probably gone early rather than late. There are rumors about contact dermatitis from photographic chemicals, but few of us know anyone who has suffered that outcome. The dangers of our work become suppressed in us. I remember hearing a muffled 'whump' when I was a graduate student, then looking out of a nearby window and seeing a refrigerator door lying on the grass three stories down from a blown-out window in the chemistry building. Later that day, we were asked to search all refrigerators for explosives, because a 'fridge' containing a can of ethyl ether had exploded (no injuries). Six cans from our floor subsequently appeared in the lone chemical fume hood, and I have never forgotten my silent concern when I removed mine. "How will I keep it from evaporating now?" Thirty-five years ago, the label on a can of ethyl ether warned against storing it in a refrigerator that was NOT explosion proof. We worked in a five story building, and there was not one such refrigerator among the thirty or so that I could count. I need not ask how many are to be found today. More, but not enough - "they are VERY expensive!" Is there one in every lab that uses ethanol at less than 4 degrees? There ought to be a continuing thread on this server about hazard and safety concerns. My self-doubt raised concerns identical to yours twenty years ago, and only a well-developed sense of self-preservation caused me to respond to an 'institutional' request for help with a very long list of reasons why I was obligated to refuse. In the process I lost favor. My first, but not only, run in with administration about job description and available resources. If my colleagues wonder why I have been so long-winded, it may be that you could use some long-winded support in your refusal to become involved in an issue for which you apparently were not hired and for which you have not been provided adequate authority to address.
Good luck Jon, your concern is not 'weird', and IT has happened a lot!
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
Hi all! I have a couple of steel wires which I want to watch in TEM. The problem is, that these wires are very fine...less than 100 microns. I want to know if anyone has any ideas about how to prepare the samples for TEM. Regards Praveena Center for advanced materials University of Massachusetts, Lowell Lowell, MA PH:978-934-3411 _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
We are having a problem with rippling when cutting 60 micron hydrogel sections. Does anyone know a way to eliminate this rippling?
Thanks, Kim
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimberly A. Riddle Florida State University tel: 850.644.6519 Biological Science Imaging Resource 119 Bio Unit I, 4370 fax: 850.644.0481 Tallahassee, FL 32306 riddle-at-bio.fsu.edu http://bio.fsu.edu/~taylor/imaging ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Help please, I'm in the market for a laser capture microdissector and would like your comments, recommendations, experiences, pro & cons, etc. of the Arcturus Pixcell II vrs. Leica's apparatus. We have seen them both, but need unbiased input from current users.
Thanks, Brett
Brett M. Connolly, Ph.D. Merck Research Laboratories Dept. of Pharmacology WP26A-3000 PO Box 4 West Point, PA 19486 ph. 215-652-2501 fax. 215-652-2075 email. brett_connolly-at-merck.com
Ladd Research has been doing tungsten filaments for many years, but we do not do the AMRAY FE filaments. We agree that it "is more than meets the eye" with these and in this case it it is probably better to go direct to the instrument manufacturer.
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955
Earl Weltmer wrote: }
} Dear Listers, } } Has anyone sucessfully done an Amray FE filament replacement for the newer } "305" guns? } } I think there is more to it than meets the eye. } } Thank You, } } Earl
Rhonda: Boy, does this bring back some really ancient memories (clear back to the late 60's!). Be films can be made the same way as C, and we evaporated Be onto glass slides in a standard evaporator (back then it was an old Kinney--but any regular evaporator will work). The *major* problem is Be toxicity, and in the vapor or dust form, Be is extremely toxic. We set up a "bell jar -in a- bell jar" system. We used a large heavy-glass beaker (probably a 2L Kimax heavy wall beaker) over the electrodes to reduce contamination of the entire unit. It is also wise to set up at least one trap in the forepump line as well as use LN2 on the diff pump. There may be some old literature out there on more specifics, but I don't know where. You may also find that some of the EM suppliers (e.g. SPI, EM Sciences, Pella, Fullam, Ladd, Tousimis, Polysciences)(sorry if I missed anyone) can supply you with the appropriate substrates. That's surely easier, altho' maybe not cheaper.
Roger
On Mon, 21 May 2001 18:07:40 -0400, Rhonda Stroud wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hello all. | | I'm looking for a suggestion for appropriate film support grids for making | EELS and EDS measurements of 1 to 10 nm particles. I would use plain | carbon, except that the particles will be a mixture of carbide, oxide, | metal and silicate phases. I need to measure the C, O, N, Si, Mg and Fe | content of the particles, so C, SiO2 or Si3N4 support films all would make my | life difficult. | | Has anyone made Be film grids? Any other suggestions? | | Thank in advance. | | Rhonda Stroud | | Research Physicist | Naval Research Laboratory | 4555 Overlook. Ave SW | Washington, DC 20375 |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I am approaching a similar need and have been looking into Boron support films. It has been done in the past although I do not have the reference handy. We hope to try making some in the next few months.
I too am interested in other options as I am interested in both Carbon, Silicon, and Boron containing samples. The Boron films will only trade conflicts and not remove the issue.
Richard Vanfleet
At 06:07 PM 5/21/01 -0400, Rhonda Stroud wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all who replied to my query on support grids.
The solution that seems best suited to my particular problem is amorphous Ge film. I'm now deciding whether to deposit the film myself, or whether to buy it.
For the curious Ge is the best because:
(1) No EDS or EELS interference with elements of interest (2) It's possible to make thin amorphous Ge films, so the background contrast will not be problem for high imaging (as apposed to Be and Al) (3) I can't use holey films because the samples will actually be collected directly on the grids during a rocket flight. Simultaneous collection on C and SiO is possible, but would make it difficult to get all info on each particle.
Rhonda Stroud
Research Physicist Naval Research Laboratory Washington, DC 20375
OK list, once again I come looking for some help. We are looking at some dirt (OK, soil) samples and I am looking for the easiest way to measure the grains(?) and the spaces between them..... Being cell and not dirt orientated I could use some help from the material sciences people. Is there a cheap or freeware program that will do this for me or do I have to basterdize my bio-imaging software (BioQuant TCW) to do this??
As usual I'm looking for the least labor intensive method. -- best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Don't go around saying the world owes you a living; the world owes you nothing; it was here first. Mark Twain [Samuel Langhornne Clemens] (1835-1910)
You can use Field Ion Microscopy sample prep techniques to prepare very sharp needles of the wires. Basically, you can dip them in a beaker and electropolish them. The liquid/air interface will preferentially polish them. You need to move the interface up and down the sample by moving either the sample or liquid. If the wires are long enough, you can float electrolyte on top of CCl4 layer and you can get two samples when the bottom part drops off. Look up some of the standard electropolishing solutions and conditions. One is based on percholoric/butylcelsolve and the other on chromic acid/acetic acid -I think.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Praveena Bhaskara [mailto:bubbyp-at-hotmail.com] Sent: Tuesday, May 22, 2001 10:36 AM To: Microscopy-at-sparc5.microscopy.com
Hi all! I have a couple of steel wires which I want to watch in TEM. The problem is, that these wires are very fine...less than 100 microns. I want to know if anyone has any ideas about how to prepare the samples for TEM. Regards Praveena Center for advanced materials University of Massachusetts, Lowell Lowell, MA PH:978-934-3411 _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
} You could call around to the small colleges in your area - as well as used } microscope vendors - for a late 1960's through mid 80's upright, retired, } transmitted fluorescence microscope from Leitz, Zeiss or B&L and Nikon. } If you are lucky, you will find one that still has its substage dark field } condenser in the nearby desk drawer. } } You could also consult geology and chemistry departments looking for a } microscope that has Leitz or Zeiss Ultropak objectives. These objectives } were/are used in epi-illuminated systems and were equipped with internal } conical mirrors that created a 'dark' field illumination. In combination } with transmitted light, such objectives were recommended for photography } of autoradiograms. } } Hope this helps. } Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: meehall-at-mac.com } Sent: Monday, May 21, 2001 9:20 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist: Which Microscope for photographs of } diatom and radiolarians } } } } Email: meehall-at-mac.com } Name: michael n maloney } } Organization: san francisco police dept } } Education: Graduate College } } Location: san francisco } } Question: i'm looking for the most appropriate microscope for fine } art photographs of diatom and radiolarian sized objects. i understand } that a true darkfield illumination source is preferable. a support } for a heavy camera is also required. I use 35 mm (Nikon F5) and a 4x5 } camera. please give me some thoughts on this } } -------------------------------------------------------------------------- } - } }
Vollenweider, H.J., Al-Be films, J. Microscopy, 16: 247(1973) from } Principles and Techniques of E M, Hayat, 3d Ed., CRC Press, 1989. }
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Rhonda Stroud } Sent: Monday, May 21, 2001 6:07 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: Be film grids? } } } Hello all. } } I'm looking for a suggestion for appropriate film support grids for making } EELS and EDS measurements of 1 to 10 nm particles. I would use plain } carbon, except that the particles will be a mixture of carbide, oxide, } metal and silicate phases. I need to measure the C, O, N, Si, Mg and Fe } content of the particles, so C, SiO2 or Si3N4 support films all would make } my } life difficult. } } Has anyone made Be film grids? Any other suggestions? } } Thank in advance. } } Rhonda Stroud } } Research Physicist } Naval Research Laboratory } 4555 Overlook. Ave SW } Washington, DC 20375 } }
I do staining of TEM thin sections of microporous polymer systems, rubber compounds and polymer based automotive coatings.
I use RuO4 mostly for staining and would like to switch to OsO4 occasionally. I use a very safe procedure in which I generate very small amounts of RuO4 vapors in a petri dish in situ. This limits problems with amounts handled, safety and minimizes disposal.
In the literature I see articles and also web sites that talk about using hot fuming nitric acid (impractical for me to keep heated), hot nitric acid (ditto) and hydrogen peroxide on "Osmium Compounds" or metal. Suggestions are made at some sites that osmium dioxide is used. OsO2 does not seem to be sold at any chemical supply houses nor is it in any EM supplier catalogs I have.
I DO NOT want to use ampoules of OsO4 or RuO4 and have them stored around the lab in sealed vials. I had an unscored ampoule of RuO4 spontaneously break once and it was not pleasant for my refrig' or me.
Question(s): Have you ever experimented with or know of any osmium compound(s) used to in situ generate osmium tetroxide vapors from one of these compounds? Is the procedure safe and does it generate small amounts of waste? What do you use as a staining indicator? Butadiene rubber or a solution of it, works for me with OsO4.
My aim here is not to find a 'cheap' way to make OsO4. Os costs a lot of money no matter what you buy or where.
Thank you for any help you can give or ideas.
Paul Beauregard PPG Industries Senior Research Associate Monroeville, PA 15146
Hi, I'm having a problem with the Coolwell Chiller (SE - Style) for my TEM. Everything is fine for a few hours then the temperature goes way up. The guys working on it checked the refrigerant level and the gauge keep fluctuating between 20 and 60. They wanted me to find out if that is normal. Any help? Thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Hi Beth, I have 3 of these SE units and know nothing about the refrigerant but have replace 2 temperature controllers because of intermittent climbing temperature. Is the red 'heat' indicator on when it overheats (make sure the bulb is good first)? When the temperature goes up try adjusting the controller down with a screwdriver. If cooling begins (blue indicator comes on) most likely the switch needs replacing and the compressor is ok. I tried cleaning the contacts but that didn't last. If the blue 'cool' light comes on but the unit's still overheating you may have a refrigeration problem.
Coolwell is out of business so to give credit where it is due here is a letter I received last year when I was looking for parts. ............................................ } We replaced the original thermostat on our Coolwell SE unit with an "ETC } Single Stage Electronic Temperature Controller" and a "1309007-044 ETC } Temperature Sensor" from Ranco (8115 U.S. 42 N., Plain City, Ohio, 43064). } The unit bolts right to the front of the Coolwell chiller and works just } fine. It works over a tempertature range of -30F to +220F and a } differential of 1F to 30F. I seem to recall that the whole shebang was } less than $100 and our campus refrigerator guy installed it with no } problems. I can fax the spec sheets to anyone who is interested. I } believe I got the idea from Maggy Piranian through this list.
} Bob } Dr. Robert R. Wise } Associate Professor of Plant Physiology } Department of Biology } University of Wisconsin Oshkosh ............................................. Regards, Jim
} X-Comment: UCONNVM.UConn.Edu: Mail was sent by sparc5.microscopy.com } Mime-Version: 1.0 } Date: Tue, 22 May 2001 16:44:12 -0500 } To: microscopy-at-sparc5.microscopy.com } From: beth-at-dogwood.botany.uga.edu (Beth Richardson) } Subject: coolwell chillers } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
Could you provide more information about what exactly you want to do? It might help people to suggest preparation techniques. do you need cross sections, look along the wires, perhaps the separation between wires?
You probably have to embed the samples in epoxy. Perhaps you can use a microtome to slice them?
Or take a whole bunch of them, embed them and then ion mill?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Praveena Bhaskara [mailto:bubbyp-at-hotmail.com] Sent: Tuesday, May 22, 2001 8:36 AM To: Microscopy-at-sparc5.microscopy.com
Hi all! I have a couple of steel wires which I want to watch in TEM. The problem is,
that these wires are very fine...less than 100 microns. I want to know if anyone has any ideas about how to prepare the samples for TEM. Regards Praveena Center for advanced materials University of Massachusetts, Lowell Lowell, MA PH:978-934-3411 _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
Can anyone point me to a source for such an objective for an old AO Microstar Series 110 microscope? I tried eBay once, but the objective didn't match the microscope requirements. Thanks.
Thanks to all who responded to my query. We had already done most of the things that were recommended. Here's an update and the apparent resolution:
According to her ophthalmologist, "Nancy's uncorrected vision has a focal point of around 6 inches. Therefore, the reticle is perfectly in focus when viewing the acellular field in the left eye, while the right does not have an image to focus on." She suggested that Nancy use corrected vision when using the microscope (e.g., contacts, implants, eyeglasses). In the mean time, I have installed a modified screening reticle that retains the outlined clear square area, but no longer includes the central circle and crossed line size comparators.
Nancy reports that the "blurring" she previously experienced has now largely gone away. In addition, she tried wearing her eyeglasses and found that the problem has disappeared entirely. She's loathe to wear her eyeglasses during microscopy, however, as they cost $400 and she doesn't want to scratch the lenses -- ocular eye guards notwithstanding.
The scientific programme for the Annual General Meeting of the Microscopical Society of Canada (June 6-8, 2001, Fredericton, NB) is established and is available at the website {http://www.unb.ca/msc2001} . Please, check the programme for a list of the topics to be presented and also note that all of the major microscopy vendors will be present to demonstrate the latest developments in confocal, light and electron microscopes. This will be the last update to the conference website.
Hello Listers, I would like to thank all of you who replied with help on finding the microspheres. I would have individually emailed those who responded, but due to a network hiccup, I lost all my email. I was fortunate enough to have hard copies of where to find some things, but no email of who to thank for the information. Have a good day, Nick
the same obj for a 60 or a 50 will also work. they are all the ao infinity series
ed sharpe archvisit for smecc
} Subj: Source of used 4X objective } Date: 5/22/01 11:41:30 PM US Mountain Standard Time } From: garygill-at-dcla.com (Gary Gill) } To: Microscopy-at-sparc5.microscopy.com } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone point me to a source for such an objective for an old AO } Microstar Series 110 microscope? I tried eBay once, but the objective } didn't match the microscope requirements. Thanks. } }
{HTML} {FONT FACE=arial,helvetica} {FONT SIZE=2} the same obj for a 60 or a 50 will also work. they are all the ao infinity {BR} series {BR} {BR} ed sharpe archvisit for smecc {BR} {BR} {BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px"} Subj: {B} Source of used 4X objective {/B} {BR} Date: 5/22/01 11:41:30 PM US Mountain Standard Time {BR} {I} From: garygill-at-dcla.com (Gary Gill) {BR} To: Microscopy-at-sparc5.microscopy.com {BR} {/I} {BR} {BR} {BR} {BR} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {BR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} -----------------------------------------------------------------------. {BR} {BR} {BR} Can anyone point me to a source for such an objective for an old AO {BR} Microstar Series 110 microscope? I tried eBay once, but the objective {BR} didn't match the microscope requirements. Thanks. {BR} {BR} Gary Gill {/BLOCKQUOTE} {BR} {BR} {/FONT} {/HTML}
I may need to replace my Argon tank on my sputter coater soon. What is the minimum purity of argon that you would recommend for use in an old Emscope SC500A sputter coater?
Thanks in Advance
Rick
Richard Portman Dep't. Biological Science University of Tulsa 600 S. College Tulsa OK 74104
I have a Philips 301 microscope that needs a new home. It is in good running condition. There are a lot of spare parts and consumables to go along with the microscope. The mercury pump has been removed. The microscope is presently up and running but will soon need to be dismantled to free up the room for another microscope. If interested please email me directly. Dismantling and shipping will be at the new owner's expense.
Debbie Lietz Electron Microscopy Technologist Biology Department, Trent University 1600 Westbank Drive Peterborough, Ontario K9J 7B8
I found this on the NET, and I hope Dr. Kiernan doesn't mind.
Re: "Disposal" of Osmium Tetroxide "Waste"
From: "J. A. Kiernan" {jkiernan-at-julian.uwo.ca}
------------------------------------------------------------------------
On Thu, 23 Nov 2000, stephen asquith wrote:
} disposal of this wonderful material but it still ...
} ... containers full of osmium in alcohol or corn oil ...
} ... what should be done with it then?
Osmium tetroxide is indeed wonderful stuff. Os is a rare
element, so disposal of used solutions should consist of
recycling, not dumping, even though osmium compounds are
not considered environmentally hazardous (Smith et al., 1978
Trace Metal in the Environment, vol 4. Ann Arbor Science
Publishers). The colourless soluble toxic tetroxide is rapidly
reduced by almost any kind of dirt to a black, insoluble dioxide,
usually in a colloidal form that's readily dispersed by moving
water if it isn't firmly stuck to the solid organic matter that
brought about the reduction.
If OsO4 slops are collected in alcohol, the os! mium (now in
the form of crude, harmless, insoluble osmium dioxide) can
be reoxidized, purified, re-reduced to pure OsO2 and stored.
OsO2 is easily re-oxidized to give a buffered solution of
osmium tetroxide (2% or less). See J Microsc 113:77-82
(1978); the procedure does involve certain hazards, so
it must be done carefully.
Recovered OsO4 can also be used to make osmeth, which is
a beautiful golden crystalline solid that contains osmium
tetroxide complexed with methenamine (= hexamethylene tetramine
or hexamine (Hanker et al., 1976 Histochemistry 49:263-291).
It costs next to nothing to make your own osmeth from
recycled OsO4, but osmeth is very expensive to buy. Osmeth
does not emit osmic fumes. When it is dissolved (in DMF
followed by dilution in an aqueous buffer) it becomes a
dilute (0.25%) working osmium tetroxide solution. I can vouch
for the excellence of home-made osmeth for post-osmication (for
EM). It may also be OK as a primary fixative or for LM stainin! g,
but I haven't encountered (personally, by anecdote or in the
literature) any use of osmeth other than for postosmication.
Perhaps someone reading this message will put me right on this.
Osmium tetroxide collected into vegetable oil could not be
recycled by the simple method cited above, and the recovery
methods used by chemists (which use apparatus etc not found
in histology labs) would be made more difficult by the presence
of the oil.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Beauregard, Paul A. } Sent: Tuesday, May 22, 2001 4:02 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Generation of in situ osmium tetroxide } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I do staining of TEM thin sections of microporous polymer systems, rubber } compounds and polymer based automotive coatings. } } I use RuO4 mostly for staining and would like to switch to OsO4 } occasionally. I use a very safe procedure in which I generate very small } amounts of RuO4 vapors in a petri dish in situ. This limits problems with } amounts handled, safety and minimizes disposal. } } In the literature I see articles and also web sites that talk about using } hot fuming nitric acid (impractical for me to keep heated), hot nitric } acid (ditto) and hydrogen peroxide on "Osmium Compounds" or metal. } Suggestions are made at some sites that osmium dioxide is used. OsO2 does } not seem to be sold at any chemical supply houses nor is it in any EM } supplier catalogs I have. } } I DO NOT want to use ampoules of OsO4 or RuO4 and have them stored around } the lab in sealed vials. I had an unscored ampoule of RuO4 spontaneously } break once and it was not pleasant for my refrig' or me. } } Question(s): } Have you ever experimented with or know of any osmium compound(s) used to } in situ generate osmium tetroxide vapors from one of these compounds? } Is the procedure safe and does it generate small amounts of waste? } What do you use as a staining indicator? } Butadiene rubber or a solution of it, works for me with OsO4. } } My aim here is not to find a 'cheap' way to make OsO4. Os costs a lot of } money no matter what you buy or where. } } Thank you for any help you can give or ideas. } } Paul Beauregard } PPG Industries } Senior Research Associate } Monroeville, PA 15146 } } } }
I am seeking assistance in finding a method to stain cadmium in paraffin embedd tissue. If such a method exists, I would very much appreciate your help.
Thank you in advance,
Diana Papoulias Fisheries Biologist, Research US Geological Survey Columbia Environmental Research Center 4200 New Haven Rd. Columbia, Missouri 65201
I use cheap, industrial grade Argon which is run through a Millipore molecular sieve. The sieve lasts for years. The cylinder of Ar costs about $20 and also lasts for years. The sieve assembly cost about $300. Virtually a one time expense.
I do the same for N2. I use industrial grade N2 ($20 per cylinder rather than 9.8 N2 ($90 per cylinder). After two years, the N2 sieve is unchanged.
gary g.
At 11:11 AM 5/23/2001, you wrote:
} Hi, } } I may need to replace my Argon tank on my sputter coater } soon. What is } the minimum purity of argon that you would recommend for use in an old } Emscope SC500A sputter coater? } } Thanks in Advance } } Rick } } } } } Richard Portman } Dep't. Biological Science } University of Tulsa } 600 S. College } Tulsa OK 74104 } } (918)631-3715
I am a photo editor and photo researcher for most texts in biology, zoology, botany, microbiology, genetics, anatomy & physiology, etc., published and I'm always searching for new sources of images to make texts more interesting, attractive, and educational. Dr. John D. Cunningham, VU Research, Inc., 46 The Flume, Amherst, NH 03031 Fax: 603-673-4738 email: john-at-visualsunlimited.com
Below is the result of your feedback form. It was submitted by (ikewlee-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 24, 2001 at 10:17:30 ---------------------------------------------------------------------------
Email: ikewlee-at-yahoo.com Name: Ike W. Lee
Organization: Anterogen
Education: Graduate College
Location: Roxbury, MA, USA
Question: Hi! I am using DAPI (Ex 364nm; Em 454nm) to trace cells during cell transplantation in animals. Two months after the transplantation when I examined the DAPI positive tissue under a light microscope, it was fluorescent not only with the DAPI filter (Ex 340-380nm; Em 420nm), but also with the two other filters (Ex 465-495, Em515-555 and, Ex 525-555, Em 590-650). Before transplantation, the cells were fluorescent with the DAPI filter only when it was checked in a tissue flask (plastic). The tissue samples were simply frozen cut without fixation. Narrow-spentrum filters of a confocal microscope were not different; in other words, DAPI positive cells were fluorescent in all confocal filters tested. My questions are: (1). Is it possible that DAPI can change its wavelength in vivo after two months? (2). Is there a way to do immunohistochemistry on these weird DAPI-positive tissue sections? (3). Are there any other methods with which I can trace cells during the two months of post-transplantation period?
Thank you.
Ike Lee Anterogen ikewlee-at-yahoo.com (617) 442-7840, ext 246
Hi, we have a problem with switching on the unit. If you turn the key, it will start, but when you release the key, it stops immediately. The Forepump does not start, we have enough air & water. All fuses check OK. Any ideas? Thank you! Peter Stolzenberg
} From memory if the instrument switches off when you release the key then the rotary pump is not running properly. You say the rotary pump does not start - do you mean that it does not start at all? If not then that is your problem - do you have power to it when the key is operated? If not then check the circuits for fuses and breakers. If so then you have a faulty motor. If the motor runs but switches off with the key - is the pump up to speed (belt slipping)? If not then there is usually a pump rotation sensor which will detect a pump fault. Has the rotation detector been knocked? Try moving it closer to the flywheel.
I don't know where you are (no affliation attached) but our local JEOL people are very good at offering advice.
Good luck, Ron
On Thu, 24 May 2001 20:59:13 EDT "Katjaalex-at-aol.com"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, we have a problem with switching on the unit. If you turn the key, it } will start, but when you release the key, it stops immediately. The Forepump } does not start, we have enough air & water. All fuses check OK. Any ideas? } Thank you! } Peter Stolzenberg }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
} } } Hi, we have a problem with switching on the unit. If you turn the key, it } will start, but when you release the key, it stops immediately. The Forepump } does not start, we have enough air & water. All fuses check OK. Any ideas? } Thank you! } Peter Stolzenberg
Peter:
I often have the same problem with my 1200EX II. Sometimes this is caused by a misalignment of the flywheel sensor on the mechanical pump. Other times it is due to insufficient air pressure. Try this: turn the power off at the main disconnect, then switch it back on; set the the key to the ON position for a few minutes before moving it to START. You may have to repeat this cycle a few times, but it always works on my instrument. Good Luck!
Ken Bart
..................................................................... ..................................................................... ............ Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
1. when starting, it is suggested you need to hold the key for a while, then release, 2. check the belt of the pump 3. check the water again, a sensor connect to oil pump will stop the machine when no water running
Yali ---------- } From: Ron Doole {ron.doole-at-materials.oxford.ac.uk} } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Jeol 1200 EX } Date: Fri, May 25, 2001, 2:14 }
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I got the maker wrong. It is Matheson Gas Products, not Millipore.
The Matheson Glass Moisture Trap (with pre-loaded sieve material) is P/N 6420-4S. The Glass Moisture Refill kit (several plastic containers of sieve material) is P/N 6420-R.
Any outfit which handles Matheson should be able to order these items for you. The sieve unit has stainless steel Swagelok fittings on each end. A really nice product. Zero maintenance.
gary g.
At 03:39 AM 5/25/2001, you wrote:
} Hi, Gary } } Can u advise the details of the Millipore thingy? } } ta } } rtch } } } } } } I use cheap, industrial grade Argon which is run through } } a Millipore molecular sieve. The sieve lasts for years. } } The cylinder of Ar costs about $20 and also lasts for years. } } The sieve assembly cost about $300. Virtually a one time } } expense. } } } } I do the same for N2. I use industrial grade N2 ($20 per cylinder } } rather than 9.8 N2 ($90 per cylinder). After two years, the N2 } } sieve is unchanged. } } } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
We have a sample of ZnSe with a thin layer of SiO2 on the surface. We would like to remove the SiO2 without damaging the ZnSe. Does anybody on the list know if ZnSe is susceptible to attack by HF? It has been suggested to me that this is possible and I was wondering if anyone has any practical experience to confirm or deny the suggestion.
Phil Flaitz IBM Microelectronics, Hopewell Junction, NY Ph.......(845) 892-3094, FAX -2003 pager - 800-352-4732, PIN# 1121 flaitz-at-us.ibm.com
I am trying to locate a histoembedder: for paraffin histology. The only one I have thus far been able to locate is one for $8500 - with way to many swanky things on it (to tell me the precices temperature of the paraffin, and under vacuum). (I don't need that - and I can't justify the school buying that kinda equipment for me). I remember using simple one - paraffin on one end, and the cold plate on the other - with only two buttons (the fewer the better, especially since this will be used in teaching undergrads).
Does anyone out there know where I can get such a machine for a reasonable price? Susan J Rehorek, Ph.D. Department of Biology Slippery Rock University of Pennsylvania PA, 16057-1326
We are looking for a microscopy objective that matches the following features:
- high N.A. (} 0.9), - water-immersion, - slim diameter (preferably { 0.7 inches or {18 mm at its end), - any magnification between 20x and 63x, - corrected for 160 mm (Nikon Measurescope MM-11), - suitable for fluorescence imaging, - preferably with a large working distance (for electrophysiology).
The lens can be new or used. Maybe you have something like that collecting dust on your shelves. Any offer is welcome.
Thanks a lot, with best regards,
Axel
Axel Blau, Ph.D. California Institute of Technology Division of Biology 156-29 1200 E. California Blvd. Pasadena, CA 91125
Why so much fuss over the argon? I was involved in the development of some of the early sputter coaters and argon was the least of our worries. We often used air when making test runs! The only problem with air is one day it may be dry the next day it may be wet so the coatings are different, well in England any way!
If some one can PROVE a difference I would be very interested to hear? If there is a problem, with different grades of argon I would be rather surprised?
Regards looking forward to some informative replies :-)
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
Phil Flaitz wrote: ============================================== We have a sample of ZnSe with a thin layer of SiO2 on the surface. We would like to remove the SiO2 without damaging the ZnSe. Does anybody on the list know if ZnSe is susceptible to attack by HF? It has been suggested to me that this is possible and I was wondering if anyone has any practical experience to confirm or deny the suggestion. ===============================================
Why could the SiO2 layer not be etched off with a reactive CF4 gas, such as CF4, using plasma etching? I would imagine that once the SiO2 layer was etched off, it would continue to etch the substrate, so some kind of control samples might have to be run to determine the exact time needed to remove the SiO2 but not etch the substrate. If one back fills to the same vacuum before starting, it is possible to get a remarkably reproducible etch on successive samples.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher, a full description of which can be found on our website indicated below.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The short answer is NO. But here is what I do know about the 305FE gun--after about two years of experience on two different FE scopes.
So I'm told, the filament is a ZrO/W unit made by Denka. The top of the filament holder is held in place by a zillion Allen head bolts. Also on the top holder plate are three pairs of large gauge wires which are the leads for the three "getters."
When the filament is replaced (factory), there is some sort of alignment procedure. This involves three small Allen head set screws on the top sides of the gun housing. Once aligned, these screws are supposedly never adjusted again. But, if during a move, and the filament assembly got really whacked out of position, it is re-aligned using these set screws. There is probably a lot more work for filament replacement than just replacing the filament. The whole electrode assembly may be cleaned or replaced (extractor, etc.).
The construction of the gun assembly is such that there is a gun isolation valve (V1) which closes off the gun from the lower column. At the top of the column is a differential pumping aperture. This is not a consumable item. Below the DPA is the column isolation valve (V3). Some of these two valves are pneumatic while others (like mine) are manual.
As-shipped, the FE gun includes a Varian 1 L/second ion pump. The gun is under high vacuum at about 8E-11 Torr. Replacement of the gun is not simply replacement of the gun assembly. It involves "conditioning" the new gun as-received via a lengthy process. The gun is well grounded to earth ground. Then, the ion pump is powered by a small power supply which provides a voltage output proportional to ion pump current. Then, the whole gun head assembly (high tension cable for all electrodes) are slowly raised to 32KV. When this is being done, the IPG (gun ion pump current) is monitored. If it starts to increase, the KV is reduced, held and then slowly increased. This process continues until 32KV is reached. If at any point, the gun head arcs, the whole process starts over. It seems to take about three hours to complete this process.
Once the conditioning is done, the gun assembly is mounted to the column. Then, vigorous pumping begins. The filament is off and V1 is still closed. Once the column vacuum reaches a safe level (typically 5E-10 Torr), the gun isolation valve is opened, vacuum checked, and the filament can be heated. It takes quite a few days for the filament to stabilize (Iext will vary for a while). Once stable, it remains very stable until it dies. Handled correctly, a gun is good for anywhere between two to four years.
Power failure, poisoning of the filament and surges can cause premature failure of the filament/gun. Right now, I am told that Amray is supporting all FE systems. The 305FE gun is the same one used on the KLA-Tencor semiconductor defect analysis tools. It is a very good gun. Extraordinarily stable. I use two double conversion UPS units (Toshiba 1400XL Plus) to protect the column and console assemblies. This helps a lot....especially here in California these days!
Under a maintenance contract, the gun is a $2K consumable. Without a contract, it is a $16K item. So I am told. I do know that it costs $2K to replace under contract since I have had to have that done one time.
gary g.
At 07:25 PM 5/21/2001, you wrote:
} Dear Listers, } } Has anyone sucessfully done an Amray FE filament replacement for the newer } "305" guns? } } I think there is more to it than meets the eye. } } Thank You, } } Earl
Listers, It appears that Pfeiffer(Balzers) Vacuum Company will no longer be supporting its older turbo pumps, like the TPH 170, 240, etc. No parts, no repairs, nothing. I received notification from Pfeiffer that this will be effective Jan. 1, 2002. Of course, they want you to buy their "new" pumps. All of the independent turbo repair shops are scrambling trying to find parts for the pumps they are repairing now, or have commitments to repair in the future. Naturally, the cost of repairs to these pumps has skyrocketed. The attraction of the use of Balzers turbo pumps in electron microscopes was the fact that this pump was in effect "self-balancing", emitting very low mechanical vibrations. Has anyone found any suitable replacement for this pump by another manufacturer? Suitable meaning not only good vibration specs but also reasonable in cost, for purchase and or to repair.
hi all someone posed a question to me after being inspected by JCAHO the } inspectors were upset they were using glut. i had never heard of a good } subsitiute for glut so i thought i would throw it out to you guys. he didn't } say if it was a labeling problem or just the fact they had glut. i have never } heard of such a thing before. } john hoffpauir } electron microscopy lab } cooper hospital camden nj
Listers, Anyone having a image of the ISI SR50A SEM(like from an old sales brochure or pic from a digital camera) that could emailed to me, it would be greatly appreciated. Please send to {mailto:gary.easton-at-scannerscorp.com} gary.easton-at-scannerscorp.com directly. Thanks in advance.
I am located on the central Atlantic coast of Florida and am considering purchase of a confocal microscope from BioRad. I would appreciate hearing, offline, from anyone in the southeastern US who has had first-hand experience (good or bad) with their equipment and service.
How about a plasma etch with CF4 + O2? That will etch the SiO2. Not sure about the other materials. You might want to run some tests on this.
gary g.
At 12:26 PM 5/25/2001, you wrote:
} We have a sample of ZnSe with a thin layer of SiO2 on the surface. We } would like to remove the SiO2 without damaging the ZnSe. Does anybody on } the list know if ZnSe is susceptible to attack by HF? It has been } suggested to me that this is possible and I was wondering if anyone has any } practical experience to confirm or deny the suggestion. } } } Phil Flaitz } IBM Microelectronics, Hopewell Junction, NY } Ph.......(845) 892-3094, FAX -2003 } pager - 800-352-4732, PIN# 1121 } flaitz-at-us.ibm.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Phil Flaitz wrote: ============================================== We have a sample of ZnSe with a thin layer of SiO2 on the surface. We would like to remove the SiO2 without damaging the ZnSe. Does anybody on the list know if ZnSe is susceptible to attack by HF? It has been suggested to me that this is possible and I was wondering if anyone has any practical experience to confirm or deny the suggestion. ===============================================
Why could the SiO2 layer not be etched off with a reactive CF4 gas, such as CF4, using plasma etching? I would imagine that once the SiO2 layer was etched off, it would continue to etch the substrate, so some kind of control samples might have to be run to determine the exact time needed to remove the SiO2 but not etch the substrate. If one back fills to the same vacuum before starting, it is possible to get a remarkably reproducible etch on successive samples.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher, a full description of which can be found on our website indicated below.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I wasn't so much concerned with the conditioning of the filament.
This process is fairly straightforward. Amray brings up the high voltage until the gun arcs causing the vacuum to deteriorate. The heaters (non-evaporative getters) are again activated to bring the pressure down again. This process is repeated until a maximum voltage of 35 KV for one hour is achieved. The external filament adjustments are two assemblies: one for X another for Y. These adjustments push on two glass rods that are internally spring loaded. The assemblies are composed of a stainless steel "cup" , a brass adjustment pin, & a small (looks like a 4-40) setscrew. The real adjustment is made on the brass pin only; the setscrew is used to limit the brass pin travel only.
I am more concerned with the initial mechanical pre-alignment of the filament itself. It appears that the filaments as sold by Denka and/or FEI come pre-aligned in it's own wehnelt assembly. This entire assembly is held in yet another "sleeve" that has another four setscrews to align it again. Seems redundant but perhaps there is a reason.
Another interesting note is that the filament construction appears to be different between the Denka & FEI filaments. This is according to their respective websites. Also the price difference is significant: FEI price is $1750.00 while Denka is $2600.00. Amray has used both vendors.
Another interesting detail is that the manufacturer's recommend that the filaments be operated at 1800 deg K. Amray runs the filaments at about 1600-1650 deg K & raises the extraction voltage to compensate. I think this would give the filament more life at the cost of some stability.
By the way, I believe there are 16 allen head screws on the top of the gun housing & the appendage pump is a 2 l/s pump.
Best Regards,
Earl
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Earl Weltmer" {earlw-at-pacbell.net} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 25, 2001 4:20 PM
Just a reminder to Listers who might be interested to attend the ASM Educational Symposium "Materials MicroCharacterization: Today and Tomorrow" next week in Oak Ridge...please check out details at http://www.korrnet.org/orcasm/. We're especially interested to have students attend, and the registration is only $20 (for students).
Larry
-- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Except for microwave fixation, which is not universally accepted, all fixatives are nasty. After all, fixatives are meant to kill cells and tissues. Coming to think of it: microwaves are nasty too, except that they are contained within the oven. And that is the take-home message for the safety officer: Any substance can be used in great safety if the correct procedures and facilities are applied. The right facilities and procedures for some substances can be too expensive, but not for the use of modest quantities of GA as in EM. A fumehood and suitable (Nitrile) gloves and fairly elementary working skills make the use of GA very save.
Where are the cases of EM related problems with GA? The few that have occurred were caused by bad facilities/ working habits or related to a sensitivity problem.
I expect that GA has a bad name in hospitals because of its use as a sterilising agent. Rooms have been sterilized by spraying GA as an aerosol and operating instruments have been bathed in large trays of GA, at times in unventilated rooms. EM people don't have such absurd applications. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, May 26, 2001 9:26 AM, "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com [SMTP:"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } hi all someone posed a question to me after being inspected by JCAHO the } } inspectors were upset they were using glut. i had never heard of a good } } subsitiute for glut so i thought i would throw it out to you guys. he } didn't } } say if it was a labeling problem or just the fact they had glut. i have } never } } heard of such a thing before. } } john hoffpauir } } electron microscopy lab } } cooper hospital camden nj
EDWARDS distributes SEIKO SEIKI magnetically levitated (active magnetic suspension of both ends) turbopumps. I am using STM-450 (450 l/sec) for 2 years and really happy with this stuff. Extremely easy to operate (actually stop/start button). Don't need any cooling (even air flow). Any orientation. And supposed to be completely vibration-free, because the rotor have no contact with "body". In case of emergency stop it uses internal electricity generator to power controller and suspension. Active suspension protects from any disbalance in the system. Does not require any technical assistance. These pumps are pricely, but EDWARDS kindly offered to me very good discount. These pumps are work-horses in industry but rare in EM community. I have no idea why. Please, let me know if you need more details.
No commercial interest, just happy user and owner.
Sergey
At 06:24 PM 5/25/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Does anyone have phase contrast and/or Nomarski optics for a 1970's Olympus Vanox microscope that they would like to sell or give away? If so please contact me.
New York Microscopical Society 30 North Mountain Avenue Montclair, NJ 07042
Bernard Friedman Memorial Workshop
Use of the Microscope September 15, 22, 29, October 6, 2001
A basic course on light microscopy which will cover the following topics: Theory of microscopy Kohler Illumination Diffraction Theory Contrast Methods Polarized light Phase Contrast Interference . . . . Hoffman contrast, Rheinberg, Dark-field & oblique Illumination, etc.
The workshop will consist of four consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of microscopy. The course instructors include Jan Hinsch of Leica, Inc., Dennis O’Leary of Micro-Optical Methods, Mary McCann of McCann Imaging, John Reffner of SensIR Technologies, Inc. and N.Y.M.S. Instructor Don O'Leary.
WHEN: September 15, 22, 29, October 6, 2001 from 10 A.M. to 4 P.M.
WHERE: 30 N. Mountain Avenue, Montclair, NJ, Phone (973) 744-0043 ( parking, accessible by public transportation, Information on car pools and transportation will be provided.)
COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: Beginners and experienced users who wish to learn more about the proper use of a microscope.
HOW: Register using the form below. Limited to the first 12 registrants. Send form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849 E-mail donoleary-at-worldnet.att.net Fax (425) 988-1415
PLEASE POST ---------------------------------------------------------------------------- ------------------- Registration Form Use of the Microscope
ISI/Topcon uses/used these pumps too. They are very nice and quite reliable. I think the reason the masses don't use them is because they are about four times more costly than the single mag bearing pumps like the Balzers. If they got close to the cost of the Balzers, they certainly would be strong contenders for replacement units. The down side is the re-wiring it would take to incorporate the Seiko units. Most of the controllers are much larger than those for the Balzers. And the in and out signals may be different.
gary g.
At 02:02 AM 5/26/2001, you wrote:
} EDWARDS distributes SEIKO SEIKI magnetically levitated (active magnetic } suspension of both ends) turbopumps. I am using STM-450 (450 l/sec) for 2 } years and really happy with this stuff. Extremely easy to operate } (actually stop/start button). Don't need any cooling (even air flow). Any } orientation. And supposed to be completely vibration-free, because the } rotor have no contact with "body". In case of emergency stop it uses } internal electricity generator to power controller and suspension. Active } suspension protects from any disbalance in the system. Does not require } any technical assistance. These pumps are pricely, but EDWARDS kindly } offered to me very good discount. These pumps are work-horses in industry } but rare in EM community. I have no idea why. Please, let me know if you } need more details. } } No commercial interest, just happy user and owner. } } Sergey } } At 06:24 PM 5/25/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Plasma etching of SiO2 with only CF4 is rather slow. The optimum mix of O2 is 10%.
Just thinking here, but you might also try Ammonium Flouride. It is a selective oxide etch and will not attack metal. At least it will not attack Aluminum. HF will attack Al. You can obtain BOE (buffered oxide etch) which has or does not have HF. This may work too. The wet etch methods may be much easier to accomplish than plasma, if you are not already set up the work and expense involved with plasma.
gary g.
At 12:26 PM 5/25/2001, you wrote:
} We have a sample of ZnSe with a thin layer of SiO2 on the surface. We } would like to remove the SiO2 without damaging the ZnSe. Does anybody on } the list know if ZnSe is susceptible to attack by HF? It has been } suggested to me that this is possible and I was wondering if anyone has any } practical experience to confirm or deny the suggestion. } } } Phil Flaitz } IBM Microelectronics, Hopewell Junction, NY } Ph.......(845) 892-3094, FAX -2003 } pager - 800-352-4732, PIN# 1121 } flaitz-at-us.ibm.com
At 08:25 PM 5/25/2001, you wrote: } Gary, } } I wasn't so much concerned with the conditioning of the filament. } } This process is fairly straightforward. Amray brings up the high voltage } until the gun arcs causing the vacuum to deteriorate. The heaters } (non-evaporative getters) are again activated to bring the pressure down } again. This process is repeated until a maximum voltage of 35 KV for one } hour is achieved.
Yes. Has to be done right or the gun will never perform correctly.
} [snip] } } Another interesting note is that the filament construction appears to be } different between the Denka & FEI filaments. } This is according to their respective websites. Also the price difference is } significant: FEI price is $1750.00 while Denka is $2600.00. } Amray has used both vendors.
I had heard that FEI may have supplied filaments. Lately, Denka seems to be more predominant. No proof of which is used more or less or not at all.
} Another interesting detail is that the manufacturer's recommend that the } filaments be operated at 1800 deg K. } Amray runs the filaments at about 1600-1650 deg K & raises the extraction } voltage to compensate. } I think this would give the filament more life at the cost of some } stability.
Each new gun comes with a comprehensive spec sheet with all parameters specified. The specs and values are taken at some particular filament current, based on each individual filament/gun. The standard practice is to reduce this current by 70mA. That is then the continuous filament current for the gun. The Vext is then adjusted to meet the gun brightness spec of 12.5nA. This takes a while (several days) to settle in on. Iext is more of a notional indicator of gun condition. It may vary a bit over time (and does vary absolutely as-delivered from gun to gun) but is quite stable until the gun starts to die. Conversely, the gun brightness value is very stable. It may vary by no more than 0.5nA or less each month.
As part of my routine qualification procedure, I take all gun status readings and the gun brightness value. A healthy gun/filament barely varies in gun brightness. It is very stable. I think the reduction in filament current was to extend the life of the filament.
} By the way, I believe there are 16 allen head screws on the top of the gun } housing & the appendage pump is a 2 l/s pump.
Ah...that explains it. I could not count all of the screws using fingers on both hands. Thanks.
Perhaps it is indeed a 2L/s pump. Still, it is a tiny little ion pump. But critical.
The Seiko pumps are not made to withstand the constant cycling from atmosphere to high vacuum that the Balzers pumps will endure.
ISI/ Topcon has valving assembly which isolates the turbo from this cycling. the Cambridge/ Leo & later Amray pump and vent the entire pumping system each time a sample is introduced.
Earl
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Sergey Ryazantsev" {sryazant-at-ucla.edu} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, May 26, 2001 7:22 AM
Retail boxed Linux (Redhat, for example) is not free. It is about the same price as Win2K.
The main problem is as you point out--application programs. While many are available for Wintel, there are few for Linux. Being an open system, Linux depends on having source code so one can compile the app for the specific Linux platform. This is unlikely to happen for industrial strength apps. They do not supply source code. And most probably, they do not support Linux.
Nevertheless, Linux with X-windows on a P-III/733 or 933 is really awesome!
gary g.
At 08:33 PM 5/21/2001, you wrote:
} I would agree that familiarity with the operating environment is important } for timely performance of server administration tasks-documentation and } advocacy groups exist for both platforms, however. It might be worth while } to become familiar with a UNIX like operating system (such as Linux) if one } has the luxury of time (I'll explain momentarily). Cost may be an } issue-Linux is free. Both OSes run on the x86 architecture. } It seems that your choice(s) of application software should be a } heavywieght factor in your decision. At the risk of over-simplification } I'll just say that most proprietary "out of the box" solutions are developed } for Windows/NT. There is a huge installed base of Wintel machines, and } there are powerful software development tools for this platform which allow } software developers to develop large complicated programs very rapidly. } Linux has the advantage that many programs written to run under the } various flavors of UNIX will compile and run. Many cutting edge scientific } applications, including image processing utilities, are developed by } researchers who are experienced coders and prefer to work under the UNIX } platform. The reason for this is partially historical-in the past only large } UNIX mainframe servers had the horsepower for image manipulation. More } often than not these software tools are available free of charge to fellow } researchers and are provided as open-source (one can view and modify the } original code to suit specialized situations). } So it really boils down to an ease of use vs. scalability/extensibility } issue with cost, and perhaps minor hardware decisions (sometimes peripherals } are difficult to configure under Linux ie. video cards, scanners etc...). } In the long run, however, time spent now learning to work with Linux may pay } off: the heart of the OS (the kernel) evolves continually in response to } the needs of users, and the kernel can be upgraded without disrupting other } parameters of the OS. Also, Linux runs on most (if not by now all)commonly } encountered chip architectures so one may have several different kinds of } machine (PC, Mac, SUN) all running the same operating system. } } Karl Garsha } www.uwm.edu/~keg } ----- Original Message ----- } } From: "Chao-Ying Ni" {cni-at-udel.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, May 18, 2001 9:36 AM } Subject: EM Lab Server OS: Linux or Windows 2000? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear colleagues, } } } } I'm going to build an EM lab server to run web and other applications, } } to share files, especially to have an interactive wed scheduling software } } to schedule and monitor usage of our 2 TEMs and 2 SEMs. Right now I have } } narrow down the platform to Linux and Windows 2000. Could you please give } } me some inputs concerning the pros and cons of these two operating } } systems? Any comments and suggestions on the hardware selection are also } } more than welcome. Thanks in advance! } } } } Chaoying Ni } } Electron Microscopy Lab } } Dept. of Materials sci. and Eng. } } University of Delaware } } Newark, DE 19716 } } cni-at-udel.edu } } } } } } } }
The question is about a server not a work station or PC. Very few applications need to be run on a server and all the ones that are needed are almost always included in the basic free distribution of Linux. Using a server for any other use is asking for trouble. Just set it up and leave it alone. I don't think anyone has actualy been at the console of my server more than a dozen time in 5 years. I just telnet in and use vi. I suppose if I had easy physical access to the box I would consider installing X windows but it way too slow over the phone line.
Almost all the Linux distributions are coming with a installation program that make it pretty simple to get things running. If you run a mailer it is potentially the greatest problem. If you are going to run a mailer I would seriously consider Linux or Unix because of the security issues with windows. With Linux in particular the security problems are quickly reported and the fixes quickly available. Who knows with windows.
Gordon Who never found a mailer I liked.
Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 } From: "Gary Gaugler" {gary-at-gaugler.com} } } Retail boxed Linux (Redhat, for example) is not free. It is about } the same price as Win2K. } } The main problem is as you point out--application programs. While } many are available for Wintel, there are few for Linux. Being an } open system, Linux depends on having source code so one can } compile the app for the specific Linux platform. This is unlikely to } happen for industrial strength apps. They do not supply source code. } And most probably, they do not support Linux. } } Nevertheless, Linux with X-windows on a P-III/733 or 933 is really awesome! } } gary g. } } } At 08:33 PM 5/21/2001, you wrote: } } } I would agree that familiarity with the operating environment is important } } for timely performance of server administration tasks-documentation and } } advocacy groups exist for both platforms, however. It might be worth while } } to become familiar with a UNIX like operating system (such as Linux) if one } } has the luxury of time (I'll explain momentarily). Cost may be an } } issue-Linux is free. Both OSes run on the x86 architecture. } } It seems that your choice(s) of application software should be a } } heavywieght factor in your decision. At the risk of over-simplification } } I'll just say that most proprietary "out of the box" solutions are developed } } for Windows/NT. There is a huge installed base of Wintel machines, and } } there are powerful software development tools for this platform which allow } } software developers to develop large complicated programs very rapidly. } } Linux has the advantage that many programs written to run under the } } various flavors of UNIX will compile and run. Many cutting edge scientific } } applications, including image processing utilities, are developed by } } researchers who are experienced coders and prefer to work under the UNIX } } platform. The reason for this is partially historical-in the past only large } } UNIX mainframe servers had the horsepower for image manipulation. More } } often than not these software tools are available free of charge to fellow } } researchers and are provided as open-source (one can view and modify the } } original code to suit specialized situations). } } So it really boils down to an ease of use vs. scalability/extensibility } } issue with cost, and perhaps minor hardware decisions (sometimes peripherals } } are difficult to configure under Linux ie. video cards, scanners etc...). } } In the long run, however, time spent now learning to work with Linux may pay } } off: the heart of the OS (the kernel) evolves continually in response to } } the needs of users, and the kernel can be upgraded without disrupting other } } parameters of the OS. Also, Linux runs on most (if not by now all)commonly } } encountered chip architectures so one may have several different kinds of } } machine (PC, Mac, SUN) all running the same operating system. } } } } Karl Garsha } } www.uwm.edu/~keg } } ----- Original Message ----- } } } From: "Chao-Ying Ni" {cni-at-udel.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Friday, May 18, 2001 9:36 AM } } Subject: EM Lab Server OS: Linux or Windows 2000? } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear colleagues, } } } } } } I'm going to build an EM lab server to run web and other applications, } } } to share files, especially to have an interactive wed scheduling software } } } to schedule and monitor usage of our 2 TEMs and 2 SEMs. Right now I have } } } narrow down the platform to Linux and Windows 2000. Could you please give } } } me some inputs concerning the pros and cons of these two operating } } } systems? Any comments and suggestions on the hardware selection are also } } } more than welcome. Thanks in advance! } } } } } } Chaoying Ni } } } Electron Microscopy Lab } } } Dept. of Materials sci. and Eng. } } } University of Delaware } } } Newark, DE 19716 } } } cni-at-udel.edu } } } } } } } } } } } } } }
Thanks for your reply. Ironically, I do use my SEIKO STM-450 for frequent air/atmosphere cycles because it installed on my vacuum evaporator. When I shop around different models (Varian, Pfeiffer and native EDWARDS) the major criterion was the ability to handle frequent air/vacuum cycles (a few per day). After many conversations and discussions with technical support teams, the SEIKO was chosen. I don't remember details (if you need, I could check), but it seems to me the pump was 1.5x more expensive than Varian with equal productivity (before discount). As I mentioned, EDWARDS give me a very nice price, so it was not so terrible expensive. Shopped for the pump I was surprised to see how companies use TPs. For example, on the Denton DV-502A 800 l/sec aca Varian M-6 diffusion pump is a standard, they offer also TP version with 150 l/sec pump. So, they installed less productive TP, perhaps to reduce the cost. I think, your impressions about the price of SEIKO based on comparison the TPs with different productivity (may be I am wrong).
As for Gary's remark about SEIKO controller, it's much smaller now. Previous models has battery installed inside controller to support active suspension during emergence shout downs. The modern models uses internal TP generator to support themselves in that case. The controller is comparable with my BALZERS controller in size. The wiring should be different for sure and perhaps SEIKO's wiring is more complicated. From another hand, SEIKO's TP do not required cooling, which is a nice feature I think. Controller, unfortunately, does not have RS-232 interface. It has a simple interface based on logistic like: contacts #1 and say #5 connected/disconnected by external relay. Using this things, you could start/stop TP and control some simply features. It was useful to me: using a few relays, electro-magnetic vacuum valves and signals from my vacuum gauge, I was able to make fully automatic system, which operated by single switch: AIR/VACUUM. Actually, I do have two more switches for safety reasons: Main and switch to shout down the TP manually in between the cycle.
Thanks to all who mentioned where in EM-world SEIKO pumps has been used (I am really happy for Hitachi and Topcon who choose this pump for their microscopes).
I am not related to EDWARDS or SEIKO and have no any interest in that companies except I am happy with their products and services.
Thanks.
Sergey.
At 01:43 PM 5/26/01 -0700, you wrote: } The Seiko pumps are not made to withstand the constant cycling from } atmosphere to high vacuum that the Balzers pumps will endure. } } ISI/ Topcon has valving assembly which isolates the turbo from this cycling. } the Cambridge/ Leo & later Amray pump and vent the entire pumping system } each time a sample is introduced. } } } } Earl } } ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Saturday, May 26, 2001 7:22 AM } Subject: Re: Balzers Turbo Pumps } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } ISI/Topcon uses/used these pumps too. They are very nice and quite } } reliable. I think the reason the masses don't use them is because they } } are about four times more costly than the single mag bearing pumps } } like the Balzers. If they got close to the cost of the Balzers, they } } certainly would be strong contenders for replacement units. The down } } side is the re-wiring it would take to incorporate the Seiko units. } } Most of the controllers are much larger than those for the Balzers. } } And the in and out signals may be different. } } } } gary g. } } } } } } At 02:02 AM 5/26/2001, you wrote: } } } } } EDWARDS distributes SEIKO SEIKI magnetically levitated (active magnetic } } } suspension of both ends) turbopumps. I am using STM-450 (450 l/sec) for } 2 } } } years and really happy with this stuff. Extremely easy to operate } } } (actually stop/start button). Don't need any cooling (even air flow). } Any } } } orientation. And supposed to be completely vibration-free, because the } } } rotor have no contact with "body". In case of emergency stop it uses } } } internal electricity generator to power controller and suspension. } Active } } } suspension protects from any disbalance in the system. Does not require } } } any technical assistance. These pumps are pricely, but EDWARDS kindly } } } offered to me very good discount. These pumps are work-horses in } industry } } } but rare in EM community. I have no idea why. Please, let me know if you } } } need more details. } } } } } } No commercial interest, just happy user and owner. } } } } } } Sergey } } } } } } At 06:24 PM 5/25/01 -0500, you wrote: } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Listers, } } } } It appears that Pfeiffer(Balzers) Vacuum Company will no longer be } } } } supporting its older turbo pumps, like the TPH 170, 240, etc. No } parts, } } } } no repairs, nothing. I received notification from Pfeiffer that this } } } } will be effective Jan. 1, 2002. Of course, they want you to buy their } } } } "new" pumps. All of the independent turbo repair shops are scrambling } } } } trying to find parts for the pumps they are repairing now, or have } } } } commitments to repair in the future. Naturally, the cost of repairs to } } } } these pumps has skyrocketed. The attraction of the use of Balzers } turbo } } } } pumps in electron microscopes was the fact that this pump was in effect } } } } "self-balancing", emitting very low mechanical vibrations. Has anyone } } } } found any suitable replacement for this pump by another } } } } manufacturer? Suitable meaning not only good vibration specs but also } } } } reasonable in cost, for purchase and or to repair. } } } } } } } } } } } } } } } } Gary M. Easton } } } } Scanners Corporation } } } } } } ------------------------------------------------------ } } } } } } Sergey Ryazantsev, Ph.D. } } } Electron Microscopy } } } Department of Biological Chemistry, School of Medicine } } } University of California, Los Angeles } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } (310) 825-1144 (office) } } } Pager: (310) 845-0248 } } } FAX: (310) 206-5272 (departmental) } } } mailto:sryazant-at-ucla.edu } } } } } } } } }
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
For most work a server does a computer being retired from the desk top is fast enough to do the job. It usually needs a bigger hard drive and more memory. Unless there are a lot of users logged on at once or a lot of large files being moved most servers don't work very hard. Build two on to go and one for a spare. If your department is like mine you should be able to come up with a few 300 MHz Pentiums and they make nice Linux servers. And use the high end machine for a better use or use the money for a better use.
I think my server is running a 133 Pentium.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} From: "Karl Garsha" {keg-at-csd.uwm.edu}
} One other concept that comes to mind, which may or may not pertain to the } individual situation we're addressing, is the issue of clustering } capability. Put simply, "clustering" provides for the linking together of } several servers over a high-speed local area network for puposes of ensuring } availability of server services in the event of a hardware malfunction, or } to balance the load on a heavily used server between multiple machines, or } both. Software to accomplish this is readily available for Linux (ie. } MOSIX: www.mosix.cs.huji.ac.il, and several others). I'm not entirely sure } if this can be accomplished under NT/2000, but I don't think so. } Clustering capability may seem extravagant considering the anticipated } use the server will recieve initially, but a couple years from now one might } be dealing with greatly expanded server load, humongous files, an image } database etc. In this case it may be attractive to hook together a couple } or few older machines rather than forcing the trusty primary server into } early retirement. } -Karl G. } } Karl Garsha } www.uwm.edu/~keg }
Hi I looking for a method for the staining of cell culture cells with ruthenium red for TEM. If have a method that works really well, I would appreciate a copy. Many thanks Manuela Palatsides Electron Microscopy Peter MacCallum Cancer Institute Locked Bag#1 A'Beckett Street Melbourne 3000
One other concept that comes to mind, which may or may not pertain to the individual situation we're addressing, is the issue of clustering capability. Put simply, "clustering" provides for the linking together of several servers over a high-speed local area network for puposes of ensuring availability of server services in the event of a hardware malfunction, or to balance the load on a heavily used server between multiple machines, or both. Software to accomplish this is readily available for Linux (ie. MOSIX: www.mosix.cs.huji.ac.il, and several others). I'm not entirely sure if this can be accomplished under NT/2000, but I don't think so. Clustering capability may seem extravagant considering the anticipated use the server will recieve initially, but a couple years from now one might be dealing with greatly expanded server load, humongous files, an image database etc. In this case it may be attractive to hook together a couple or few older machines rather than forcing the trusty primary server into early retirement. -Karl G.
**************************************************************************** **************************************************************************** ****** Karl Garsha www.uwm.edu/~keg
Does anyone have any suggestions for sources of good B&W (TV-rate) monitors?
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Dear Sir or Madam; Does anyone suggest suitable acids for etching and polishing of Cu-Mn(25%)-Al(11%) alloys and Cu-Mn(25%)-Zn(11%),Cu-Mn(25%) and Mf and Af temperature alloys? Thanks for your interested.
********************************** ********************************** ** Research.Asst.ERDEM YASAR ** ** University of Kirikkale ** ** Department of Physics ** ** TEM LABORATORY ** ** 71450 KIRIKKALE/TURKEY ** ** erdem.yasar-at-physics.org ** ** yasar-at-science.ankara.edu.tr ** ** yasar-at-turkuaz.kku.edu.tr ** ********************************** **********************************
Below is the result of your feedback form. It was submitted by (megaprpf-at-juno.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, May 28, 2001 at 03:31:58 ---------------------------------------------------------------------------
Email: megaprpf-at-juno.com Name: Robert Bauman
Organization: Amarillo College
Education: Undergraduate College
Location: City, State, Country
Question: How long does one leave TEM polymer in a oven to harden it enough so that it can be sectioned on an ultramicrotome?
Electronic Surplus houses tend to get these... I have even heard stories of them being scrapped since the market is soft for them and some times dealers get a large lot.... They will put a few in the store and send the rest out for scrap.
thanks Ed Sharpe archivist for SMECC
{ { Subj: Re: B&W monitors Date: 5/28/01 10:43:01 AM US Mountain Standard Time From: {A HREF="mailto:COURYHOUSE"} COURYHOUSE {/A} To: {A HREF="mailto:ldm-at-risc4.numis.nwu.edu"} ldm-at-risc4.numis.nwu.edu {/A} To: {A HREF="mailto:microscopy-at-sparc5.microscopy.com"} microscopy-at-sparc5.microscopy.com {/A}
Electronic Surplus houses tend to get these... I have even heard stories of them being scrapped since the market is soft for them and some times dealers get a large lot.... They will put a few in the store and send the rest out for scrap.
I am considering purchase of a cryo ultramicrotome from Micro Star Technologies. I would appreciate comments from anyone who has had experience (good or bad) with this equipment and service.
Thanks in advance
Carmen Bracho Centro de Microbiologia, IVIC Phone/fax: + 58 212 504 17 59 e-mail: cbracho-at-ivic.ve
We routinely get inspected by JCAHO and CAP (College of American Pathologists). We've never had a problem with the inspectors. All EM labs, that I know of, including clinical ones, use glut for fixation. Find out just what the problem was--labeling, handling, or the fact that it was used at all.
Sara
On Fri, 25 May 2001 JHoffpa464-at-aol.com-at-sparc5.microscopy.com wrote:
} Date: Fri, 25 May 2001 18:25:35 -0500 } From: JHoffpa464-at-aol.com-at-sparc5.microscopy.com } To: Microscopy-at-sparc5.microscopy.com } Subject: subsitiute for glut } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } hi all someone posed a question to me after being inspected by JCAHO the } } inspectors were upset they were using glut. i had never heard of a good } } subsitiute for glut so i thought i would throw it out to you guys. he } didn't } } say if it was a labeling problem or just the fact they had glut. i have } never } } heard of such a thing before. } } john hoffpauir } } electron microscopy lab } } cooper hospital camden nj } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I've been contacted by a retired biologist who runs a large photo service for publishers who need images. He says he can use a wide variety of subjects & he's particularly interested in the newer microscopies - "STEM, atomic force, molecular probe, fluorescent and confocal, etc." Please contact him directly if you have some good stuff to share:
} Photographs utilizing all types of microscopy are solicited for use in } science textbooks at all levels, children's science books, trade books, field } guides, aquarium and museum exhibits, TV programs (e.g., NOVA, Discovery } Channel), and other educational publications. Credit and appropriate payment } is given for each usage. Contact Dr. John D. Cunningham, VU Research/Visuals } Unlimited, 46 The Flume, Amherst, NH 03031 for information. Ph: (603) } 673-4653 Fax: (603) 673-4738 Email: john-at-visualsunlimited.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
} From: "L. D. Marks" {ldm-at-risc4.numis.nwu.edu}
} Does anyone have any suggestions for sources of good B&W } (TV-rate) monitors? }
Your video production people on campus should be able to give you sources for them as well and may have surplus monitors on hand. If they can't help the local TV stations should be able to help.
Look in the yellow pages under security systems if you want to buy new ones locally.
If price is a problem get in touch with some local amateur radio operators and ask about the fellows working with TV broadcasting. They should have some or know where to get some for very reasonable prices. They will probably still have the university inventory stickers on them from the university auctions.
Yahoo shopping show a number of them as well http://search.shopping.yahoo.com/search/all?P=all&is=1&p=%22B%26w+monitor% 22&cl=dmiyg
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
Assuming that you are looking for the composite video monitor (video and sync. signals mixed, as in common household TV set- all in single wire, and 75 Ohm input impedance, though security monitor usually has input switch for high and low impedance): your most economical bet will be security hardware vendors. You will find many in your local ph. book. Try also these: www.supercircuits.com or (800)335-9777 ; and www.radioshack.com or (800)291-6515. Both have catalogs online or available by mail. Use monitor with vertical and horizontal hold controls for the best compatibility with your signal source.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: L. D. Marks {ldm-at-risc4.numis.nwu.edu} To: Microscopy List {microscopy-at-sparc5.microscopy.com}
Does any one know of the banding problem in the 2 Quadrant solid state Back scatter electron image in JEOL 6335 FESEM? We see vertical bands mixed with the BEI image. The service engineer says that this is because of ground leakage current. We measured the ground potential of 200 mv peak to peak w.r.t the chassis. Although we try to believe that this is because of inefficient sheilding, there could be other reasons too. Request you all to suggest what could be wrong.
Thanks well in advance Prabhakar M
GE India Technology Centre ________________________________________________ M.Prabhakar PSM Group John F. Welch Technology Centre Export Promotion Industrial Park, Phase II Hoodi Village, Whitefield Road Bangalore-560 066, India Tel.No. : 080-8412050-69 Extn. : 2574 Fax. : 080-8412111 E-mail: M.Prabhakar-at-geind.ge.com
As part of the introduction of a quality system in our laboratory I have been asked to write SOP's for the operation of the SEM and X-Ray analysis system. I am not exactly sure what is required for this and wondered if anyone else has been through this process. I would welcome advice as to how much information is required on the actual operation of the equipment and what other information is needed.
Regarding a previous request of mine about "Remote Access to SEM" I would like to thank the people who responded. I had hoped to reply before now but the process is ongoing. I hope to be able to report a successful solution in the near future.
Many Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2.
Tel: 353-1-6081820 Fax: 353-1-6770438 Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
I was affraid from that information, as I have a dozen from these types of turbo pumps in service in our lab, so I have asked Pfeiffer in France what about this thing. So today the answer is clear : in France they continue to do maintenance on both type. For repear, only rotor and motor for 170 are not avaible, but for 240 all is avable, service and parts. They said it seems to be an "US" problem. Probably, the reprentativ want to sell new pumps... But its seems to be according to the general commercial politic of this manufatcurer in the last years.
Three years ago, I hade a TPU 330 to repear (the dual rotor type, the rotor was in marmalade), and they propose me an exchange with an new 240 for the price of repear. They could repear it, but it didn't pay.
As an other type of pump, I use the Varian V70 and V250.They are very reliable, the controller gives much informations than can be used to drive it carrefully. But as my use is for UHV surface analysis and plasma coating, vibration is not a criterion, and I have some doubt about their level for microscopy. And it is loudy... But I think that Leo use it on the Gemini 1530/50.
We have also a Seiko Seiki magnetic bearing model, whitch sees often air (vacuum evaporator for organic compounds). The pump is reliable, but not the electronic. And you must send both back for repear. The after sale is long (6 month one time), done by Edwards.
It's the same thing with a Pfeiffer TPU520-M ( full dynamic magnetic bearings) on UHV STM. The pump itself is reliable, very good on vibration aspect, the electronic less good. But with this type of pump, we have an other limiation : the vessel is on an air sustended table, and if you knock in the vessel, the pump will act as a gyroscope and make an emergency stop. It doesn't like too much that kind of stop. It would be the same with a SEM. OK, it is not something to do, to knock a STM or a SEM in work !
I have now bought an new Pfeiffer (TMU 261), but it is unpacked today. I'll say something about it in a few month. The controller is not compatible with the former series (TPU/H 110,170,240,260,33,520), and vice versa.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Listers, It appears that Pfeiffer(Balzers) Vacuum Company will no longer be supporting its older turbo pumps, like the TPH 170, 240, etc. No parts, no repairs, nothing. I received notification from Pfeiffer that this will be effective Jan. 1, 2002. Of course, they want you to buy their "new" pumps. All of the independent turbo repair shops are scrambling trying to find parts for the pumps they are repairing now, or have commitments to repair in the future. Naturally, the cost of repairs to these pumps has skyrocketed. The attraction of the use of Balzers turbo pumps in electron microscopes was the fact that this pump was in effect "self-balancing", emitting very low mechanical vibrations. Has anyone found any suitable replacement for this pump by another manufacturer? Suitable meaning not only good vibration specs but also reasonable in cost, for purchase and or to repair.
BSE amplifiers typically run at high gains. That makes them susceptible to many kinds of interference. Yes, it could be a grounding problem (one would hope that the manufacturer's service rep could take care of that). It could also be induced through the pre-amplifier and the cabling. Coaxial cables only partially shield their signal wire. If there is anything near the pre-amp or cabling that is capable of producing EMF (including portions of the instrument itself), it will probably be picked up by the system.
Try turning off or moving any ancillary equipment around the SEM. This would include any external vacuum gauges, EDS systems and monitors and any motors (chillers or mechanical pumps). Try re-routing the cabling to the BSE detector and pre-amp. Once the source is identified, you can take steps to minimize the problem.
One question does have to be asked, though. BSE imaging is a method to distinguish between areas of differing atomic number averages. The closer those atomic number differences, the more gain you will use to differentiate them. If you are placing unrealistic expectations on your system by expecting large image contrast from small differences in average atomic numbers, you may well be pushing the capabilities of your system. Can you provide more information regarding the materials you are seeking to image?
On Monday, May 28, 2001 11:26 PM, M, Prabhakar (CORP, GEITC) [SMTP:M.Prabhakar-at-geind.ge.com] wrote: } } } Does any one know of the banding problem in the 2 Quadrant solid state Back } scatter electron image in JEOL 6335 FESEM? } We see vertical bands mixed with the BEI image. The service engineer says } that this is because of ground leakage current. } We measured the ground potential of 200 mv peak to peak w.r.t the chassis. } Although we try to believe that this is because of inefficient sheilding, } there could be other reasons too. Request you all to suggest what could be } wrong. } } Thanks well in advance } Prabhakar M } } GE India Technology Centre } ________________________________________________ } M.Prabhakar } PSM Group } John F. Welch Technology Centre } Export Promotion Industrial Park, Phase II } Hoodi Village, Whitefield Road } Bangalore-560 066, India } Tel.No. : 080-8412050-69 Extn. : 2574 } Fax. : 080-8412111 } E-mail: M.Prabhakar-at-geind.ge.com } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
I believe that everyone on this listserver has already received this request.
On Monday, May 28, 2001 2:54 PM, Caroline Schooley [SMTP:schooley-at-mcn.org] wrote: } } } I've been contacted by a retired biologist who runs a large photo service } for publishers who need images. He says he can use a wide variety of } subjects & he's particularly interested in the newer microscopies - "STEM, } atomic force, molecular probe, fluorescent and confocal, etc." Please } contact him directly if you have some good stuff to share: } } } Photographs utilizing all types of microscopy are solicited for use in } } science textbooks at all levels, children's science books, trade books, field } } guides, aquarium and museum exhibits, TV programs (e.g., NOVA, Discovery } } Channel), and other educational publications. Credit and appropriate payment } } is given for each usage. Contact Dr. John D. Cunningham, VU Research/Visuals } } Unlimited, 46 The Flume, Amherst, NH 03031 for information. Ph: (603) } } 673-4653 Fax: (603) 673-4738 Email: john-at-visualsunlimited. } } Caroline } } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
We have the problems with our JEM-4000EX: the tremblihg of the electron beam spot. Short history. The filament in our microscope work more 800 hours. High tension there was stable at 400kV. But the electron beam began to tremble. The beam spot on the screen aperiodic tremble in one direction with frequency from 1 to 20 Hz. Sometimes, fluctuations stop and all there was OK. But with each daytime situation grew worse. In addition increase the beam current : there was 123.9 (400Ë×), became 124.4. At 300kV there was 92.2, became 92.7. We have to change the filament. Here we have clean up of the Wehnelt and first anode as well have changed the silicon grease on insulator. After change the filament made the pumping of the microscope 2 days. Then we made all operation according to instruction "High voltage generation after filament exchange": annealing of the filament at îô=0kV etc. As the HT at 400 kV there was unstable ( the fluctuations of vacuum meter SIP-GUN) we use the N2 glow discharge conditioning at 150kV. Then once again we made the filament annealing at HT=0kV. High tension became stable at 400 kV, but the beam spot on the screen begins aperiodic tremble at 370 kV and above (as before change the filament). The training of HT for 3 days a situation did not perfect. We have opened the gun once again. On insulator and on Wehnelt have not the traces of disruption. The checkup of the filament has shown that crystal LaB6 stand orderly, but fastening its to threads of incandescence (do not know what this alloy) on the one hand flake away. We have changed the filament on new. And have done once again all procedures, described above. The situation did not change, but even grew worse. The beam current increase to 127mA at 400Ë×. The fluctuations of beam spot begin at 350kV. I have the following questions. 1. How to check with than is the trembling of the electron spot : contamination in Gun or Column of the microscope or instability of a electronic circuits? I think that the high tension is stable since in diffraction mode the size of the caustic is stable and not changed. 2. The new filaments are kept in original packaging, but not in vacuum. Time of keeping more than 10 years. Can be the filaments degrade for this time? 3. The electron spot trembling are fixed on the oscilloscope picture in check point CH (HV tank) (In position OPERATE). In position TEST of the ACCEL VOLTAGE switch the oscilloscope picture is stable (direct line). As it is generated HT in position TEST? Looking forward to hear some recommendation which can help us. In Russia we have not regularservice of JEOL Ltd. Thank you very much in advance. I am not currently part of the list, so please mail to me directly. Best regards, Anton Gutakovskii mailto:gut-at-thermo.isp.nsc.ru
Does any one know of the banding problem in the 2 Quadrant solid state Back scatter electron image in JEOL 6335 FESEM? We see vertical bands mixed with the BEI image. The service engineer says that this is because of ground leakage current. We measured the ground potential of 200 mv peak to peak w.r.t the chassis. Although we try to believe that this is because of inefficient sheilding, there could be other reasons too. Request you all to suggest what could be wrong.
Thanks well in advance Prabhakar M
GE India Technology Centre ________________________________________________ M.Prabhakar PSM Group John F. Welch Technology Centre Export Promotion Industrial Park, Phase II Hoodi Village, Whitefield Road Bangalore-560 066, India Tel.No. : 080-8412050-69 Extn. : 2574 Fax. : 080-8412111 E-mail: M.Prabhakar-at-geind.ge.com
I have had the pleasure (cough, cough) of writing SOP's (we call them SLP's in QS9000, ISO equivalent for the automotive industry, for system level procedures). Basically, what you need to do is write a set of procedures that you can live with. The amount of detail is really up to you. If you have experienced people that do not change very much, then they don't have to be very long. If you have a high turnover and people need to be trained frequently, then you can use the SLP as a training aid. They do come in handy when a person does some tasks infrequently such as cleaning wehnelts, changing filaments, or mixing chemicals.
What you will be required to do however is follow the procedures that you write down. If you don't, you can get written up for a non-conformance and that could be job-threatening. The more specific you are, the less "play" you have in using the instruments. Therefore, you should sprinkle in phrases such as "depending on operator experience," "suggested" and "recommended" liberally throughout the document. For example, the following is an excerpt from the section of the details to the procedures for our TEM SLP (it is in boldface and all caps to get the attention.:
Procedures: I: NOT ALL OF THE STEPS LISTED IN THIS PROCEDURE ARE REQUIRED TO BE FOLLOWED. OPERATOR EXPERIENCE AND TRAINING ARE USED TO JUDGE WHICH STEPS ARE APPLICABLE AND IN WHAT ORDER TO BEST APPLY THESE STEPS.
I suggest writing things down that are very specific with respect to service, calibration procedures, and things that you need to do to protect the microscope. These can be done in an outline form. Anything that you do routinely you can put down specifically. Other types of work, you should be more general in what you say so that you don't get pinned down on doing the work in a regimented way. You have to give yourself some flexibility to use the power of the microscope. What you should realize is that you are the expert, -not the examiner. All he/she is going to do is determine whether the paper trail on your work is there and if you follow your procedures. So, CYA!
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Colin Reid [mailto:creid-at-tcd.ie] Sent: Tuesday, May 29, 2001 1:20 AM To: MSA Listserver
Hi,
As part of the introduction of a quality system in our laboratory I have been asked to write SOP's for the operation of the SEM and X-Ray analysis system. I am not exactly sure what is required for this and wondered if anyone else has been through this process. I would welcome advice as to how much information is required on the actual operation of the equipment and what other information is needed.
Regarding a previous request of mine about "Remote Access to SEM" I would like to thank the people who responded. I had hoped to reply before now but the process is ongoing. I hope to be able to report a successful solution in the near future.
Many Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2.
Tel: 353-1-6081820 Fax: 353-1-6770438 Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
We have a client who wants to look at how calcium is localized in processed cheese samples. She thinks it may be in the fats and lipids, but that's part of the research question. My question is: does anyone have any knowledge of how to prepare cheese samples for EDS analysis in an SEM?
My inclination would be to do a gluteraldehyde fix and skip the heavy metal post-fixes to avoid overwhelming the Ca signal with Os and UA signals, but then this might leave a lot of lipid content unfixed and susceptible to extraction during dehydration. We don't have an environmental scope, unfortunately.
Any thoughts would be very welcome.
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Hi, we have a problem with switching on the unit. If you turn the key, it will start, but when you release the key, it stops immediately. The Forepump does not start, we have enough air & water. All fuses check OK. Any ideas? Thank you! Peter Stolzenberg
Several years ago, during preparation of zirconium/zirconium oxide cross section samples for TEM, we observed that the metal sometimes became completely converted to Zr hydride during argon ion micromilling. The hydriding effect in reactive metals such as Zr and Ti during ion milling had been recognized and studied in some depth earlier as described by Carpenter et al. in the J. Micros. Soc. of America, Vol. 1, No. 4 (1995) 175-184. Although their study identified hydrocarbon contamination as the cause, the operating conditions in our case pointed to water vapor in the argon. Ordinary grades of Ar including "prepurified" grade actually have a rather high dewpoint. Among several measures taken to eliminate the hydriding artifact, changing to ultrahigh purity grade had the greatest, immediately evident effect. We also installed inline molecular sieve/charcoal filters in the gas feed system ( most gas suppliers sell these filters for removing specific contaminants) and made it standard practice to use an LN2 cold trap in the ion mill. Cooling the sample also would have been preferable, but the ion mill we were using did not allow this.
Larry Thomas
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto:Larry.Thomas-at-pnl.gov
} } I use cheap, industrial grade Argon which is run through } } a Millipore molecular sieve. The sieve lasts for years. } } The cylinder of Ar costs about $20 and also lasts for years. } } The sieve assembly cost about $300. Virtually a one time } } expense. } } } } I do the same for N2. I use industrial grade N2 ($20 per cylinder } } rather than 9.8 N2 ($90 per cylinder). After two years, the N2 } } sieve is unchanged. } } } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
First place I look is: Hayat, M.A.(1989), Principles and Techniques of Electron Microscopy Biological Applications (3rd Ed.), CRC Press, Boca Raton, FL, USA (ISBN: 0-8493-7111-2).pp285-289
Second is: Hayat, M.A.(1993), Stains and Cytochemical Methods, Plenum Press, NY, NY, USA, (ISBN: 0-306-44294-9), pp305-318. (mention of cultured cells), chemistry and (inter)actions.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Palatsides, Manuela } Sent: Sunday, May 27, 2001 6:56 PM } To: microscopy list } Subject: TEM: Ruthenium Red } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi } I looking for a method for the staining of cell culture cells with } ruthenium } red for TEM. If have a method that works really well, I would appreciate } a } copy. } Many thanks } Manuela Palatsides } Electron Microscopy } Peter MacCallum Cancer Institute } Locked Bag#1 } A'Beckett Street } Melbourne 3000 } } Telephone: 03 96561244 } Fax: 03 96561411 } Email: m.palatsides-at-pmci.unimelb.edu.au } } } } } }
The best SOP's you are likely to find are those prepared by SEM'ers involved with Forensic analysis. This is a quick and dirty suggestion, however, and is not intended to exclude any others who have confronted the issue in their labs. Quality control in industry also likely requires rigorous SOP's, and I am sure their are others. The Forensic folks talk to one another, and they can be easily approached on the NET.
State of Virginia, USA, Forensic Science Lab might be a possibility. http://www.dfs.state.va.us/
I searched GOOGLE for "Standard Operating Procedures SEM EDS and found among the many the following. http://www.plu.edu/~benhamsr/page20.html - probably what your leader would like as a start.
Hope this helps,
Fred
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Colin Reid } Sent: Tuesday, May 29, 2001 1:19 AM } To: MSA Listserver } Subject: Writing SOP's for SEM/X-Ray Analysis } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } As part of the introduction of a quality system in our laboratory I have } been asked to write SOP's for the operation of the SEM and X-Ray analysis } system. I am not exactly sure what is required for this and wondered if } anyone else has been through this process. I would welcome advice as to } how much information is required on the actual operation of the equipment } and what other information is needed. } } Regarding a previous request of mine about "Remote Access to SEM" I would } like to thank the people who responded. I had hoped to reply before now } but the process is ongoing. I hope to be able to report a successful } solution in the near future. } } Many Thanks. } } Best wishes, } } Colin } } Colin Reid, } Electron Microscope Unit, } Trinity College Dublin, } Dublin 2. } } Tel: 353-1-6081820 } Fax: 353-1-6770438 } Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm } } }
Here is a TEM prep that I use on confluent cell monolayers to demonstrate the integrity of tight junctions.
30 minute fix in 2.5% glutaraldehyde in 0.1 M Sorensen's buffer, pH 7.4 and containing 0.1 % ruthenium red.
Two 3 minute rinses in 0.1 M Sorensen's buffer
Post fix for 15 minutes in 1 % osmium in Sorensen's buffer containing 1 % ruthenium red.
Two 3 minute rinses in buffer.
Quick rinse in ddH20 to remove salts.
en block stain for 15 minutes in aqueous 3 % uranyl acetate.
Dehydrate 3 minutes each in 50, 70, 90, and one change of 100 % EtOH.
Infiltrate with 3:1, 1:1, 1:3 EtOH:Epon for 20 minutes each.
Change to full strength epon for 1 hour.
Change to new full strength epon for 1 hour.
Polymerize
I hope this helps.
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Mon, 28 May 2001, Palatsides, Manuela wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi } I looking for a method for the staining of cell culture cells with ruthenium } red for TEM. If have a method that works really well, I would appreciate a } copy. } Many thanks } Manuela Palatsides } Electron Microscopy } Peter MacCallum Cancer Institute } Locked Bag#1 } A'Beckett Street } Melbourne 3000 } } Telephone: 03 96561244 } Fax: 03 96561411 } Email: m.palatsides-at-pmci.unimelb.edu.au } } } } }
Look at the magnetic sensor on the flywheel of your Roughing Pump. The positioning of this unit is quit sensitive. Good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone 512/282-5507 FAX 512/280-0702
Sustaining Member - MICROSCOPY SOCIETY OF AMERICA
----- Original Message ----- } From: {"Katjaalex-at-aol.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, May 29, 2001 10:41 AM
Hi Mark,
Yes you are correct. In my mind if you are coating for SEM when you will be running at magnifications of 15,000X upwards you will have almost certainly have hidden or destroyed the specimen with a coating that looks gold!
The heat that you report is understandable with long coating times I believe it is fairly easy to understand.
1. We irradiate a target with argon (or air) gas molecules. 2. This heats up the surface as well as causing it to be sputtered, as well as "polishing" the surface. 3. A polished hot surface will radiate heat - what sit opposite to this target but your (now) hot specimens!
I have always argued that for an operator to fully understand a sputter coating unit it takes about an hour of useful tests.
1. Work at a 5cm target to specimen distance. 2. Cover the base plate with a piece of photocopy paper (will contain gas and possible moisture just like a specimen) place a small weight (an SEM stub is ideal) on the paper to stop it blowing away. 3. Coat for several minutes at 20mA with a high voltage coater (1kV to 3kV with a variac type control) or for a minute with a low voltage coater at 10mA ( has a deposition control not a kV control). 4. Look at the paper to see the coating distribution - almost certainly there will be good and poor areas - note the position of the good areas. Also note how round the stub the coating is not so good, this is typical of areas around a specimen, they will affect the coating. 5. Using a similar piece of paper coat for multiples of 30 seconds on fresh pieces of paper, for up to 3 minutes. Using the criteria set out above for current and distance. 6 For typical SEM operations look for the coating that is the first grey color that you obtain the colors will be - pink - light blue - darker blue - grey - darker grey with gold tinges and eventually gold. 7. Place two small nuts (as in nuts and bolts) on copy paper on top of each other, to create a hole about 1/4 inch high by 1/8th inch wide. 8. Run your coater and try every route at your disposal to get gold or your preferred metal down the hole.
Once you have found out alI these answers I think you will know a great deal more about your coater. You should now know -
a) What I call the target spots in your system, the positions for the highest coating efficiency. b) The coating criteria for a reasonable coat and minimum metal structure. c) The procedure for really difficult specimens with "tooth brush type" structures. {Have you tried this one, its a bit of a surprise to find that you cut off the gas and run at the best possible vacuum - think why?}
Never expect a sputter coating alone to overcome charging, if as an operator you insist on using 15kV plus on difficult to coat specimens expect trouble. To help think about tilting towards the Everhart-Thornley detector to increase the BSE contribution to the image as BSE are not troubled by the charge. With very complex specimens coat with the sample tilted 45 degs in one direction then 45 in the other then use the "hole" technique as described above to drive the metal into deeper areas. But as I always say we are as electron microscopists "scientists" and scientists should experiment :-)
Hope this helps, have fun!
Steve Chapman Senior Consultant Protrain For professional training wide wide in SEM, TEM and EDX www.emcourses.com
When in the past I have looked at calcium/phosphorus distribution in cheese it has been on frozen samples, using a Hexland (now Gatan) cryo-stage on our SEM. Providing the fracture plane was OK, this worked generally well and we achieved some quite interesting digital maps.
"Tindall, Randy D." {TindallR-at-missouri.edu} on 29/05/2001 15:35:27
To: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com} cc:
Hi Listers,
We have a client who wants to look at how calcium is localized in processed cheese samples. She thinks it may be in the fats and lipids, but that's part of the research question. My question is: does anyone have any knowledge of how to prepare cheese samples for EDS analysis in an SEM?
My inclination would be to do a gluteraldehyde fix and skip the heavy metal post-fixes to avoid overwhelming the Ca signal with Os and UA signals, but then this might leave a lot of lipid content unfixed and susceptible to extraction during dehydration. We don't have an environmental scope, unfortunately.
Any thoughts would be very welcome.
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
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I think this is a job for Low-temperature SEM. Any attempt to do a chemical fix risks extraction of soluble ions from the aqueous phase of the cheese. Chris
} Hi Listers, } } We have a client who wants to look at how calcium is localized in processed } cheese samples. She thinks it may be in the fats and lipids, but that's } part of the research question. My question is: does anyone have any } knowledge of how to prepare cheese samples for EDS analysis in an SEM? } } My inclination would be to do a gluteraldehyde fix and skip the heavy metal } post-fixes to avoid overwhelming the Ca signal with Os and UA signals, but } then this might leave a lot of lipid content unfixed and susceptible to } extraction during dehydration. We don't have an environmental scope, } unfortunately. } } Any thoughts would be very welcome. } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } }
To All, Sorry about the post, It was meant to be private. Tom Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility View expressed are my own.
Your question about 3D reconstruction based on 2D sections was forwarded to my desk by an associate who is a member of your group.
There are many reconstruction packages on the market. One of the ones I am aware of is the VoxBlast by VayTek (http://www.vaytek.com/VoxBlast.html).
Another I have seen is called SpyGlass but I do not have a URL reference for that one.
Best of luck.
Matt
Matthew M. Batchelor Applications Engineer ------------------------------------ Meyer Instruments, Inc. 1304 Langham Creek, Suite 235 Houston, TX 77084 Video Microscopy Imaging Specialists P - 281-579-0342, F - 281-579-1551 E-Mail = mmb-at-meyerinst.com Home Page = http://www.meyerinst.com/
If the banding is very regular, I'd be inclined to think its an electronic problem. If its irregular, then I suspect a charging effect - is your specimen well grounded? Try the BEI with just a blank mount. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, May 29, 2001 11:19 PM, M, Prabhakar (CORP, GEITC) [SMTP:M.Prabhakar-at-geind.ge.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does any one know of the banding problem in the 2 Quadrant solid state Back } scatter electron image in JEOL 6335 FESEM? } We see vertical bands mixed with the BEI image. The service engineer says } that this is because of ground leakage current. } We measured the ground potential of 200 mv peak to peak w.r.t the chassis. } Although we try to believe that this is because of inefficient sheilding, } there could be other reasons too. Request you all to suggest what could be } wrong. } } Thanks well in advance } Prabhakar M } } GE India Technology Centre } ________________________________________________ } M.Prabhakar } PSM Group } John F. Welch Technology Centre } Export Promotion Industrial Park, Phase II } Hoodi Village, Whitefield Road } Bangalore-560 066, India } Tel.No. : 080-8412050-69 Extn. : 2574 } Fax. : 080-8412111 } E-mail: M.Prabhakar-at-geind.ge.com
Dear LISTERS-FRIENDS, Thank your very much to everyone for your kind replies on my problems with electron miocroscope JEM-4000EX.I hope that your recommendations will be use to us. Please find enclosed more details about our problems with JEM- 4000EX. I check the stability îô by oscilloscope in óî at îô=280kV ( the filament is off). The oscillogram is direct line, i.e. HT is stable. Then I raise îô to 300kV. At first HT=300kV is stable. The beam current is 93.5 mkA. In 3-5 min the beam current increase to 94.2 mkA and a saw-tooth pulses appear on oscilloscope picture. The amplitude of the pulse is ~0.2V, time length is about 5 mS and on-off time ratio is 25-40 mS. When I reduce the îô to 280kV the saw-tooth pulses present at oscilloscope picture already 20-30 min, but afterwards disappear and îô becomes stable. When I raise HT to 300kV again then the situation is repeated. The vacuum in GUN is 1.2x10-5Pa. The fluctuation of the vacuum meter is half partition of the meter skale. Previously vacuum was 6-7x10-6Pa. Have You some new ideas on this cause? Thank you very much in advance. Best regards Anton mailto:gut-at-thermo.isp.nsc.ru
Hi all, I am writing to ask for advise and assistance in locating sources of used equipment. I am starting a new position in the Fall at a smallish undergrad campus in the U of Wisconsin system and am trying to setup a basic microscopy center for both teaching and research. My funds are very limited but I really need certain items, particularly a cryostat. Any help would be greatly appreciated!!!! Please contact me directly at my current address, lyden.11-at-osu.edu. Thanks in advance.
Tim Lyden, Ph.D. Research Scientist Heart and Lung Institute Ohio State University
I have experienced banding on BSE imaging in the past. It is almost always caused by 50 (or 60) Hz interference from the electrical mains due to inadequate earthing of cable shielding. Whether the problem is due to mains frequency pickup can be seen by running at different scan rates - the bands on the image should become narrower with slower scan rates.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
} } On Tuesday, May 29, 2001 11:19 PM, M, Prabhakar (CORP, GEITC) } [SMTP:M.Prabhakar-at-geind.ge.com] wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Does any one know of the banding problem in the 2 Quadrant solid state Back } } scatter electron image in JEOL 6335 FESEM? } } We see vertical bands mixed with the BEI image. The service engineer says } } that this is because of ground leakage current. } } We measured the ground potential of 200 mv peak to peak w.r.t the chassis. } } Although we try to believe that this is because of inefficient sheilding, } } there could be other reasons too. Request you all to suggest what could be } } wrong. } } } } Thanks well in advance } } Prabhakar M } } } } GE India Technology Centre } } ________________________________________________ } } M.Prabhakar } } PSM Group } } John F. Welch Technology Centre } } Export Promotion Industrial Park, Phase II } } Hoodi Village, Whitefield Road } } Bangalore-560 066, India } } Tel.No. : 080-8412050-69 Extn. : 2574 } } Fax. : 080-8412111 } } E-mail: M.Prabhakar-at-geind.ge.com }
Colin: In line with Scott's comments, (having had to do this same thing when moving from academia to industry), the basic thing is to use the KISS principle (i.e. keep it simple s----). We have used the approach to refer to the equipment manual(s) as much as possible for actual instrument functions. The major things expected in our SOPs (for the FDA) are: equipment log (user, date, sample IDs, etc); equipment maintenance procedures (routine, emergency, repair, etc, including chain of command, notification--ie out of service posting--, signature of service personnel, and so on). Calibration (standards, maintenance, frequency, ...) is also another biggie. Above all, make it something you can live with, something that is not too detailed and, most importantly, useful.
Roger On Tue, 29 May 2001 06:19:55 +0100, Colin Reid wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hi, | | As part of the introduction of a quality system in our laboratory I have | been asked to write SOP's for the operation of the SEM and X-Ray analysis | system. I am not exactly sure what is required for this and wondered if | anyone else has been through this process. I would welcome advice as to | how much information is required on the actual operation of the equipment | and what other information is needed. | | Regarding a previous request of mine about "Remote Access to SEM" I would | like to thank the people who responded. I had hoped to reply before now | but the process is ongoing. I hope to be able to report a successful | solution in the near future. | | Many Thanks. | | Best wishes, | | Colin | | Colin Reid, | Electron Microscope Unit, | Trinity College Dublin, | Dublin 2. | | Tel: 353-1-6081820 | Fax: 353-1-6770438 | Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Manuela: Good responses from Fred and Dorothy. The Hayat references are certainly more accessible than the original papers by John Luft. The work by Luft, however, demonstrated the need for a very good source of ruthenium red (available from most EM supply houses nowadays). While not a terribly fussy procedure, use of a poorly defined RR will give variable results. I wish you the best. Roger On Mon, 28 May 2001 08:56:49 +1000, Palatsides, Manuela wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | | Hi | I looking for a method for the staining of cell culture cells with ruthenium | red for TEM. If have a method that works really well, I would appreciate a | copy. | Many thanks | Manuela Palatsides | Electron Microscopy | Peter MacCallum Cancer Institute | Locked Bag#1 | A'Beckett Street | Melbourne 3000 | | Telephone: 03 96561244 | Fax: 03 96561411 | Email: m.palatsides-at-pmci.unimelb.edu.au | | | | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
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I sent a response directly to the original sender of this message, but received a "not a subscriber" error message in return. If I get a viable address, I'll resend. Roger On Mon, 28 May 2001 09:36:55 -0500, megaprpf-at-juno.com wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Below is the result of your feedback form. It was submitted by | (megaprpf-at-juno.com) from | http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, May | 28, 2001 at 03:31:58 | --------------------------------------------------------------------------- | | Email: megaprpf-at-juno.com | Name: Robert Bauman | | Organization: Amarillo College | | Education: Undergraduate College | | Location: City, State, Country | | Question: How long does one leave TEM polymer in a oven to harden it | enough so that it can be sectioned on an ultramicrotome? | | --------------------------------------------------------------------------- |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Hello, I work in the Central Imaging Facility at the University of Vermont. WE have had a request to look for bacteria on Tygon tubing using the SEM. THe request came from a dentist. I was wondering if you could lend some insight as to how we might go about doing this or if you know of someone that might be able to help. Thank you Nicole
The origin can be magnetic, also, I had a similar problem with my JEOL 840, which uses an external BSE preamp that sits on the table beside the column. Maybe your model does, too.
I have also an optical microscope, whose lamp was being powered by a small transformer, with mutiple taps on the secondary, which was within 6" or so from the BSE preamp.
I had discounted this as the cause, because the transformer was always switched to "off" whenever a BSE image was being collected, so, I thought, no magnetic field.
However, relocating the transformer eliminated the banding. I guess there was sufficient magnetic field radiated from the transformer even when there was no current being drawn from the secondary.
So check that there are no AC-current-carrying leads nearby.
cheers
rtch
} } Does any one know of the banding problem in the 2 Quadrant solid } state Back scatter electron image in JEOL 6335 FESEM? We see } vertical bands mixed with the BEI image. The service engineer } says that this is because of ground leakage current. We measured the } ground potential of 200 mv peak to peak w.r.t the chassis. Although } we try to believe that this is because of inefficient sheilding, } there could be other reasons too. Request you all to suggest what } could be wrong. }
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I have a paper I could send you that may be of interest. It is titled "High Resolution Scanning Electron Microscopy of Milk Products: A New Sample Preparation Procedure" by McManus, McMahon and Oberg. Food Structure, Volume 12 (1993) pp 475-482. The "new" part may not be so relevant as the paper is from 1993.
Some of the materials they worked with were mozzarella, swiss and cream cheeses. This really isn't my area of expertise, but I have the paper here because they used our Ion Beam Sputtering system as part of the process. If you would like a copy, I will be pleased to send it over to you.
Best regards-
David
"Tindall, Randy D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Listers, } } We have a client who wants to look at how calcium is localized in processed } cheese samples. She thinks it may be in the fats and lipids, but that's } part of the research question. My question is: does anyone have any } knowledge of how to prepare cheese samples for EDS analysis in an SEM? } } My inclination would be to do a gluteraldehyde fix and skip the heavy metal } post-fixes to avoid overwhelming the Ca signal with Os and UA signals, but } then this might leave a lot of lipid content unfixed and susceptible to } extraction during dehydration. We don't have an environmental scope, } unfortunately. } } Any thoughts would be very welcome. } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Sat, 26 May 2001, Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Retail boxed Linux (Redhat, for example) is not free. It is about } the same price as Win2K. } } The main problem is as you point out--application programs. While } many are available for Wintel, there are few for Linux. Being an } open system, Linux depends on having source code so one can } compile the app for the specific Linux platform. This is unlikely to } happen for industrial strength apps. They do not supply source code. } And most probably, they do not support Linux. } } Nevertheless, Linux with X-windows on a P-III/733 or 933 is really awesome! } } gary g. } } } At 08:33 PM 5/21/2001, you wrote: } } } I would agree that familiarity with the operating environment is important } } for timely performance of server administration tasks-documentation and } } advocacy groups exist for both platforms, however. It might be worth while } } to become familiar with a UNIX like operating system (such as Linux) if one } } has the luxury of time (I'll explain momentarily). Cost may be an } } issue-Linux is free. Both OSes run on the x86 architecture. } } It seems that your choice(s) of application software should be a } } heavywieght factor in your decision. At the risk of over-simplification } } I'll just say that most proprietary "out of the box" solutions are developed } } for Windows/NT. There is a huge installed base of Wintel machines, and } } there are powerful software development tools for this platform which allow } } software developers to develop large complicated programs very rapidly. } } Linux has the advantage that many programs written to run under the } } various flavors of UNIX will compile and run. Many cutting edge scientific } } applications, including image processing utilities, are developed by } } researchers who are experienced coders and prefer to work under the UNIX } } platform. The reason for this is partially historical-in the past only large } } UNIX mainframe servers had the horsepower for image manipulation. More } } often than not these software tools are available free of charge to fellow } } researchers and are provided as open-source (one can view and modify the } } original code to suit specialized situations). } } So it really boils down to an ease of use vs. scalability/extensibility } } issue with cost, and perhaps minor hardware decisions (sometimes peripherals } } are difficult to configure under Linux ie. video cards, scanners etc...). } } In the long run, however, time spent now learning to work with Linux may pay } } off: the heart of the OS (the kernel) evolves continually in response to } } the needs of users, and the kernel can be upgraded without disrupting other } } parameters of the OS. Also, Linux runs on most (if not by now all)commonly } } encountered chip architectures so one may have several different kinds of } } machine (PC, Mac, SUN) all running the same operating system. } } } } Karl Garsha } } www.uwm.edu/~keg } } ----- Original Message ----- } } } From: "Chao-Ying Ni" {cni-at-udel.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Friday, May 18, 2001 9:36 AM } } Subject: EM Lab Server OS: Linux or Windows 2000? } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear colleagues, } } } } } } I'm going to build an EM lab server to run web and other applications, } } } to share files, especially to have an interactive wed scheduling software } } } to schedule and monitor usage of our 2 TEMs and 2 SEMs. Right now I have } } } narrow down the platform to Linux and Windows 2000. Could you please give } } } me some inputs concerning the pros and cons of these two operating } } } systems? Any comments and suggestions on the hardware selection are also } } } more than welcome. Thanks in advance! } } } } } } Chaoying Ni } } } Electron Microscopy Lab } } } Dept. of Materials sci. and Eng. } } } University of Delaware } } } Newark, DE 19716 } } } cni-at-udel.edu } } } } } } } } } } } } } }
The best way would probably be with the use of a cryo stage on your SEM. The process will preserve your samples in their most natural state. In fact, Polaron uses a photo micrograph of cheese in some of its cryo marketing materials.
We are the U.S. distributors for The Polaron Range, including their cryo stage. Cryo systems are also available from other manufacturers such as Gatan (formerly the Oxford System), Bal-Tec and Emmitech, to name a few.
I hope this helps.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, May 29, 2001 5:57 PM To: Tindall, Randy D. Cc: microscopy-at-sparc5.microscopy.com
I think this is a job for Low-temperature SEM. Any attempt to do a chemical fix risks extraction of soluble ions from the aqueous phase of the cheese. Chris
} Hi Listers, } } We have a client who wants to look at how calcium is localized in processed } cheese samples. She thinks it may be in the fats and lipids, but that's } part of the research question. My question is: does anyone have any } knowledge of how to prepare cheese samples for EDS analysis in an SEM? } } My inclination would be to do a gluteraldehyde fix and skip the heavy metal } post-fixes to avoid overwhelming the Ca signal with Os and UA signals, but } then this might leave a lot of lipid content unfixed and susceptible to } extraction during dehydration. We don't have an environmental scope, } unfortunately. } } Any thoughts would be very welcome. } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } }
I would agree with Scott's reply, make sure that you can live with the SOP that you write. We are in the process of writing an operation SOP for our SEM. We started with the operation manual and had an experienced user write our SOP. We then had everyone in our department review the SOP. Some of the non-users had some good suggestions for improvement. Finally, we had a new user who was learning the SEM use the SOP and make suggestions. This sounds like a lot of work, but after all that input we think we have a document that we can work with and that will help us train future users. We did include some statements that allowed for use of scientific judgement, but kept these to a minimum.
Good Luck,
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
"Colin Reid" {creid-at-tcd.ie To: "MSA Listserver" {Microscopy-at-sparc5.microscopy.com} } cc: Subject: Writing SOP's for SEM/X-Ray Analysis 05/29/01 12:19 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
As part of the introduction of a quality system in our laboratory I have been asked to write SOP's for the operation of the SEM and X-Ray analysis system. I am not exactly sure what is required for this and wondered if anyone else has been through this process. I would welcome advice as to how much information is required on the actual operation of the equipment and what other information is needed.
Regarding a previous request of mine about "Remote Access to SEM" I would like to thank the people who responded. I had hoped to reply before now but the process is ongoing. I hope to be able to report a successful solution in the near future.
Many Thanks.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2.
Tel: 353-1-6081820 Fax: 353-1-6770438 Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
Call Belair at 1-800-783-9424 they sell refurbished cryostats.
Diane Miller Consultant Service 503-627-0130
----- Original Message ----- } From: "Tim Lyden" {lyden.11-at-osu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 30, 2001 2:31 AM
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
Dear colleagues, I am trying to do ultrathin sectioning work of ZnO crystals, which size is about 2-3microns(L) and 1-2microns(W). I am really a freshman on TEM, so I hope to get help on this work. Any suggestions for the choosing of embedding media and following procedures will be greatly appreciated. Thank you in advance.
Dujin Wang Max-Planck-Institute for Polymer Research Ackermannweg 10, Mainz D-55128 Germany
I am working in small particle contacting with TEM and I am interested in interpreting the contacting boundary structure using Bollmman's geometric model.
Can you offer me any information about the computerised way or software which can graphically show the CSL misroientation solution of grain boundary dislocation structures for cubic structure by methods based on that of Bollmann.
Thank you,
Yiming ---------------------------------
Yiming Yao
Microscopy and Microanalysis Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
I take it from your message that the manual for the vacuum system is independent of the manual for the XL-30. I think I would tend to believe FEI. We operate our LaB6 routinely at 1 x 10-6.6 to 1 x 10-6.8 mbar. Our slightly poorer vacuum probably does result in somewhat shortened LaB6 life, but it's the best vacuum our aging ion pump can manage, and we rarely have "down" time because of vacuum problems. In a perfect vacuum, of course, the LaB6 filament would last a very, very long time, (theoretically); and the poorer the vacuum, the shorter the filament life. But I think in this case FEI is right - your vacuum system manual may be "too restrictive" for real-world operation. I would consider 6 x 10-7 an admirable vacuum, at least in our instrument. And, of course, FEI will be happy to sell you a new LaB6 when you need one ;-). If your new XL-30 is able to do its own "bake-out" routinely, then by all means use this function as often as you feel is necessary. And do keep an eye on the state of the vacuum - you may notice changes for the better or worse as time goes on. Good luck.
F.C. Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
----- Original Message ----- } From: "FABBRI" {fabbri-at-cigssrv1.unimo.it} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 31, 2001 4:55 AM
At the US Department of Energy Albany Research Center we review our SOPs annually so that changes and suggestions can be documented and included. I tell people that if they see needed changes and suggestions for the SOPS let me know so these can be incorporated. Everyone who operates our SEMs must read and sign the SOP for that equipment.
It is my opinion that an SOP does not replace the manufacture's operator and service manuals, software online help and operators training. But I also realize that some management want very detailed SOPS so I am thankful for scanners and word processors.
} Hi all, I am writing to ask for advise and assistance in } locating sources of used equipment...
} My funds are very limited...
Tim,
I am very much in the same boat. I am setting up an SEM lab out of my own pocket - no funding from a university, corporation, or government. I have had good success by constantly checking the auction sites www.labx.com and www.ebay.com. Also, Duniway Stockroom of Mountain View, CA (www.duniway.com 800-446-8811) stocks a lot of supplies that you may need at very reasonable prices.
The most important thing about the auction sites is patience. It takes a while for some items to come up, but over the past six months, I have seen two sputter coating units, lots of vacuum pumps, a critical point dryer, lots of ultrasonic cleaners, and zillions of light microscopes. Plenty of other stuff, too. I'm pretty sure that I've seen a cryostat, but since I wasn't looking for one, I didn't pay any attention to it.
If you are patient and persistent, my experience indicates that you can buy things at about 10 to 20 per cent of the original price. I am currently sitting at my desk looking at a Polaron critical point dryer that just arrived this morning. I bought it for $100. I am awaiting the arrival of a Polaron sputter coater that I bought for $465. I bought a Varian ionization gauge controller for $165 and the gauge tube (also Varian) for $35. I bought the cable to connect the two from Duniway Stockroom for $89. Varian wanted $190. The electron microscope itself came from none other than your new employer, the University of Wisconsin, at no cost other than truck rental and fuel.
The usual auction site disclaimers apply - Make sure that you know exactly what you are bidding on - get model numbers and check the specs. Find out whether or not the unit is known good, might be good, is good for parts only, etc. Be willing to let go of an item if the bidding gets too high - another item of the same type will eventually appear. Don't let you emotions get the better of you. Set your budget limit and stick to it. Read the auction rules. eBay and LabX operate differently. On eBay, they use proxy bidding so that you don't have to watch the bidding so closely, but someone can steal an item from you at the last second. LabX works more like a real auction where you have to monitor the bidding closer, but if someone dumps in a bid at the last second, the auction is extended for a period of two minutes to allow you time to respond.
Good luck. The search can be fun, if a little frustrating at times. I wish you well in your new position.
Since the SEM is new, it should be still on warranty. However, get FEI representative's advice in writing on company letterhead to protect yourself. If you think the XL30 is not performing to specifications, call them in to find out why it isn't.
Greg Lum Lab Manager EM Facility San Francisco State University
---------- Forwarded message ----------
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
In my files I find the following companies that deal in used and rebuilt equipment:
Capovani Brothers Inc. http://www.capovani.com Tel: 518-346-8347
E. McGrath inc. Tel 508-744-3546
Good luck, WCB -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Look in Thompson, S.W.(1966), "Selected [1600+ pages!!!!] Histochemical and Histopathological Methods", Thomas Pub., Springfield, IL, USA
Chap 9. "Microscopic Histochemical Methods for the Demonstration of Minerals" p.p.. 587-589 Sodium rhodizonate (CAS 523-21-7). Primary method for Lead, but Cd is listed a minor contributor under certain conditions. While the Lead complex is violet and very strong, Cd complex is 'brown-red' and not as strong a Rx. Al Rx's are pH dependent. This is a classic "in-your-hands" cytochemical method, but it is for LM. Just when you think all is lost in the past, a little light appears, as follows.
ALSO, Cd is bound by oxime (also 8-hydroxyquinoline[CAS: 148-24-3]). The latter is used to chelate Cd. See, Yoshizuka, M, McCarthy, K.J. Kaye, G.I., and Fujimoto, S.(1990), Cadmium toxicity to the cornea of pregnant rats: Electron Microscopy and X-ray microanalysis, Anat. Rec., 227: 138.(from Hayat,M.A.(1993), STAINS AND CYTOCHEMICAL METHODS, Plenum, N.Y., NY, pp.375-76). The reference in Hayat is a bit sketchy, but the resolution for you is about right. Fortunately, the primary record should be readily available.
BUT, seems to me if there is sufficient Cd for there to be a question, a good thin section (not from paraffin) would show it using STEM or at lower res (you CAN use a paraffin section!) using SEM w/EDS or better res w/WDS. Some college/university/company should have one of these and be willing to help. With the right combination of a heavy metal 'stain' (lead or uranium as long as it doesn't exchange with the Cd!), you should be able to get a good tissue map with backscatter and then a great overlay of that with the X-Ray map of the Cd distribution. Without the reference, I can't back this memory up, but it seems to me that if you have an animal with suspected Cd poisoning, one of the best places to look is the testis in which it concentrates (I think!). Sort of like looking for Iodine in the thyroid.
PubMed Search:
27. Kawahara A, Yoshizuka M, Hirano T, Ohsato K, Fujimoto S. Related Articles
Cadmium toxicity in perinatal rat hepatocytes: electron microscopy, X-ray microanalysis, and morphometric analysis.
Exp Mol Pathol. 1990 Oct;53(2):180-90.
PMID: 2261947 [PubMed - indexed for MEDLINE]
28: Yoshizuka M, McCarthy KJ, Kaye GI, Fujimoto S. Related Articles
Cadmium toxicity to the cornea of pregnant rats: electron microscopy and x-ray microanalysis.
Anat Rec. 1990 May;227(1):138-43.
PMID: 2368924 [PubMed - indexed for MEDLINE]
Since you work at *.gov, this should not be an insurmountable problem.
Good Luck and hope this helps,
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Diana M Papoulias } Sent: Wednesday, May 23, 2001 7:48 PM } To: microscopy-at-sparc5.microscopy.com } Subject: cadmium stain } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am seeking assistance in finding a method to stain cadmium in paraffin } embedd tissue. If such a method exists, I would very much appreciate your } help. } } Thank you in advance, } } Diana Papoulias } Fisheries Biologist, Research } US Geological Survey } Columbia Environmental Research Center } 4200 New Haven Rd. } Columbia, Missouri 65201 } } voice mail: 573-876-1902 } e-mail: Diana_Papoulias-at-usgs.gov } fax: 573-876-1896 } } }
A colleague wishes to get guidance and information about coupling his video camera to his compound microscope.
He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet lens mount. He wants to use it on a Zeiss Axiophot.
I am used to simple C-mount adapters, the bayonet mount is the ringer for me.
According to his research Sony offers an adapter, HRT045ENG12, that they say is used to mount this camera to a microscope. His question is 'Does the Sony adapter substitute for those offered by Zeiss for this purpose, or is the Sony adapter used to convert the bayonet mount to something compatable with the adapters offered by Zeiss?'
Finally, back in the old days, I remember learning that one could use a simple T-mount to attach a 35 mm camera to a microscope, but that it was not as good as having an eyepiece camera because it lacked the ocular lens. Is the same true of mounting a video camera? I have always just used simple, lenless C-mounts and it has seemed fine. If we are looking for ultimate image quality, should we be using a more sophisticated adapter?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
} Hi all, I am writing to ask for advise and assistance in } locating sources of used equipment...
} My funds are very limited...
Hi Tim,
I have started new molecular biology and tissue culture labs. Several vendors (e.g., Fisher Scientific) provide discounts for new lab startups. For example, I called Fisher and they sent me some forms to complete. That was it. Whenever I placed an order, I told the operator about my discount. The discounts vary with each vendor, and they can save you a substantial sum. If you must purchase new equipment and supplies, you might want to ask about these.
Good luck!
(Disclaimer: All opinions are my own, not those of my employer.)
Kristine L. Danowski Dow Chemical Company Corporate R&D Analytical Sciences 1897 Building Midland, MI 48667
The wisest man I ever knew taught me something I never forgot. And although I never forgot it, I never quite memorized it either. So what I'm left with is the memory of having learned something very wise that I can't quite remember. -- George Carlin
Has anyone here familiar with any attempts to achieve critical point drying of a water saturated sample without the use of an intermediate fluid, i.e., by using the critical point of water itself at 374 oC/3212 PSI?
I'm interested in looking at clay samples. I am concerned that acetone will cause structural rearrangement of the clay particles because of dissimilar physical and electrical properties between acetone and water. Unlike biological specimens, clay would not care the least about a temperature of 374 degrees, as it does not begin its dehydroxylization process until over 500 degrees. I work in an oil field related industry where we have ready access to equipment that would consider 5000 PSI to be "light duty" so an appropriate pressure bomb would not be difficult to construct.
Some questions:
1) All CPD devices that I have seen have a window that allows you to observe the state of the meniscus. Is this essential, or can I assume that the process will go as physics says it should without actually observing the meniscus?
2) I do not want to place the clay samples in water and allow the water to provide pressure as it is heated, as any water in contact with the clay will start to decompose the sample. Can I use an external pressure source (high pressure air, for example) to keep the pressure inside the bomb high enough to avoid evaporation of any water until the critical point is reached? Would it suffice to simply build a little platform that would keep the clay sample above the water level?
3) Am I nuts for even considering this? What am I missing?
Thanks for your help.
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
The following figures is a generalization to some degree, since I do not know what brand of LaB6 cathode you are using. LaB6 cathodes manufacturers advertise expected lifetime (hours) under vacuum conditions of 10-7 mbar or better. When vacuum deteriorates closer to 10-6 mbar range, cathode lifetime will be reduced about twice. Generally, LaB6 cathodes are not recommended for use in vacuum (even intermittently) worse than 10-6 mbar, as cathode lifetime reduces dramatically (exponentially). Especially for LaB6 cathodes with metallic heater (versus Carbon one).
XL30 is a good instrument and must achieve much better vacuum than 6 x 10-7 mbar, continuously. Also, baking out may only improve vacuum once after initial pump down in a system with negligible leaks. The fact that periodic bake-out improves vacuum temporarily, may point to another condition, such as micro leak(s). When parts of the gun/column are heated, materials do expand, and leak(s) may (partially) close for awhile. Or, a substantial amount of outgasing material is present in the UHV area. Or, vacuum readout may be incorrect. In which case everything might be just fine!
All this is just a suggestion. A number of other scenarios possible, with a number of proper troubleshooting techniques. If your SEM is still under warranty, the solution is up to your service representative. His inspiration is up to you.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: FABBRI {fabbri-at-cigssrv1.unimo.it} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
G'day Bruce, What is wrong in going the other way and simply dry your sample using a small vacuum chamber?? High temperature and high pressure spell trouble to me. Regards JVN
Bruce Girrell wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Has anyone here familiar with any attempts to achieve critical point drying } of a water saturated sample without the use of an intermediate fluid, i.e., } by using the critical point of water itself at 374 oC/3212 PSI? } } I'm interested in looking at clay samples. I am concerned that acetone will } cause structural rearrangement of the clay particles because of dissimilar } physical and electrical properties between acetone and water. Unlike } biological specimens, clay would not care the least about a temperature of } 374 degrees, as it does not begin its dehydroxylization process until over } 500 degrees. I work in an oil field related industry where we have ready } access to equipment that would consider 5000 PSI to be "light duty" so an } appropriate pressure bomb would not be difficult to construct. } } Some questions: } } 1) All CPD devices that I have seen have a window that allows you to observe } the state of the meniscus. Is this essential, or can I assume that the } process will go as physics says it should without actually observing the } meniscus? } } 2) I do not want to place the clay samples in water and allow the water to } provide pressure as it is heated, as any water in contact with the clay will } start to decompose the sample. Can I use an external pressure source (high } pressure air, for example) to keep the pressure inside the bomb high enough } to avoid evaporation of any water until the critical point is reached? Would } it suffice to simply build a little platform that would keep the clay sample } above the water level? } } 3) Am I nuts for even considering this? What am I missing? } } Thanks for your help. } } Bruce Girrell } Microline Technology Corp. } 2397 Traversefield Dr. } Traverse City, MI 49686 } http://www.microlinetc.com } } (231) 935-1585 (Voice) } (231) 922-5099 (FAX) } bigirrell-at-microlinetc.com
-- John Nailon Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
True, when the vacuum is marginal or when "accidents" occur, baking can save the day. Although, there are limitations and eventually a cathodes will fail. My point is that the performance of the established LaB6 brands are fairly similar under ideal conditions, but Kimball's single center post design is intrinsically more robust and will tolerate more abuse before failure. I consider this is an established fact and is not an impression clouded by my obvious commercial interest. Disclaimer: ProSciTech supplies Kimball LaB6 (not to N America) Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, May 31, 2001 9:43 PM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Dr. Fabbri - } } I take it from your message that the manual for the vacuum system is } independent of the manual for the XL-30. I think I would tend to believe } FEI. We operate our LaB6 routinely at 1 x 10-6.6 to 1 x 10-6.8 mbar. Our } slightly poorer vacuum probably does result in somewhat shortened LaB6 life, } but it's the best vacuum our aging ion pump can manage, and we rarely have } "down" time because of vacuum problems. In a perfect vacuum, of course, the } LaB6 filament would last a very, very long time, (theoretically); and the } poorer the vacuum, the shorter the filament life. But I think in this case } FEI is right - your vacuum system manual may be "too restrictive" for } real-world operation. I would consider 6 x 10-7 an admirable vacuum, at } least in our instrument. And, of course, FEI will be happy to sell you a new } LaB6 when you need one ;-). } If your new XL-30 is able to do its own "bake-out" routinely, then by all } means use this function as often as you feel is necessary. And do keep an } eye on the state of the vacuum - you may notice changes for the better or } worse as time goes on. } Good luck. } } F.C. Thomas } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada } } } } ----- Original Message ----- } } From: "FABBRI" {fabbri-at-cigssrv1.unimo.it} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, May 31, 2001 4:55 AM } Subject: XL30 - LaB6 Vacuum system } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } We are using a quite new FEI XL-30 LaB6 SEM. } } It's a long time the we are trying to understand the } } normal working conditions of the vacuum system. Looking } } at what is written in the operating manual, our vacuum system is not able } } to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). } } To do this we need to bakeout the gun frequently ( weekly )and are able } } to work in that safety ( { 2x10-7 )condition only for few hours. } } FEI says that the info on the manual are too restrictive (!!) and that } } we can work safetly with vacuum levels {6 x10-7 baking out every two } weeks. } } } } Any thoughts would be very welcome. } } Thank you very much in advance. } } } } Best regards } } P.L. Fabbri } } } } --------------------------------------------- } } Dr. Pier Luigi Fabbri } } C.I.G.S. - Centro Interdip. Grandi Strumenti } } Universita di Modena } } via Campi 213/A - 41100 Modena, Italy } } Tel +39-059-2055231 / +39-059-370551 } } Fax +39-059-370551 } } E-mail: fabbri-at-mail.cigs.unimo.it } } --------------------------------------------- } } } } } } } } } }
AutoQuant makes a nice program called AutoVisualize. It will take 2D slices and make them into a 3D image.
gary g.
At 03:59 PM 5/29/2001, you wrote:
} Kevin (and others), } } Your question about 3D reconstruction based on 2D sections was forwarded to my } desk by an associate who is a member of your group. } } There are many reconstruction packages on the market. One of the ones I am } aware of is the VoxBlast by VayTek (http://www.vaytek.com/VoxBlast.html). } } Another I have seen is called SpyGlass but I do not have a URL reference } for that } one. } } Best of luck. } } Matt } } Matthew M. Batchelor } Applications Engineer } ------------------------------------ } Meyer Instruments, Inc. } 1304 Langham Creek, Suite 235 } Houston, TX 77084 } Video Microscopy Imaging Specialists } P - 281-579-0342, F - 281-579-1551 } E-Mail = mmb-at-meyerinst.com } Home Page = http://www.meyerinst.com/
Gary Gaugler, Ph.D. Optical Reflections 7970 Twin Rocks Rd Granite Bay, CA 95746-8111 916.791.8191 916.791.8186 fax
Disclaimer: I am an authorized dealer for this and other photographic and software products.
Hello all, Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. Thanks. Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
We have a LaB6 equipped ESEM installed in early 2000. It is still using the original filament which has now logged over 1000 hours. Although we have had various problems with the instrument vacuum levels have not been part of it. Gun vacuum is in the range 2.0 - 2.6 x10-7.
We bake it out overnight every time air is admitted to the gun for cleaning or repair. We are in the process of installing a high vac valve between the ion pump and the gun to save on bake outs at pressured times.
Sounds like you have a 'microleak' - get the agents onto it while you are still under warranty ! Baking out every two weeks might appeal to the purists who want to ensure 'ultimate vacuum' but it seems like overkill - unless it is compensating for a latent problem.
Another point that may be of interest - we have found that the instrument (with mixed lo and hivac usage) will last about three months before it starts getting wobbly ie gun instability and uncorrectable astigmatism. This is an indicator to clean the Wehnelt - itself no mean task since you are cleaning off an entirely transparent but totally nonconductive layer of Lanthanum (?) from the inside of the cap - checking for continuity with a multimeter as you clean. Once baked out after this process it performs like a star once again ! Thus far we are cleaning with polish, but are investigating less exhausting chemical cleaning methods !
The local service agents for FEI/Philips Anaspec-at-icon.co.za have assisted us a great deal in coming to terms with the idiosyncrasies of a LaB6 equipped ESEM - a concept which has given us some interesting moments.
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0) 33 260 5155 Fax +27 (0) 33 260 5776 website via:http://www.nu.ac.za/department/default.asp?dept=cemunp Email:bruton-at-nu.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
} } } "FABBRI" {fabbri-at-cigssrv1.unimo.it} 05/31/01 09:55AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
Access to cryoSEM or ESEM would save a lot of work.
(Without any real experience of clays) I would be inclined to just air dry the sample at 40 deg. C in an oven.
The meniscus is required when replacing the intermediate fluid, to prevent exposing the sample, since you would not have one I guess it is irrelevant.
Dave
On Fri, 01 Jun 2001 10:06:22 +1000 John V Nailon {J.Nailon-at-mailbox.uq.edu.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } G'day Bruce, } What is wrong in going the other way and simply dry your sample using a } small vacuum chamber?? } High temperature and high pressure spell trouble to me. } Regards } JVN } } Bruce Girrell wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Has anyone here familiar with any attempts to achieve critical point drying } } of a water saturated sample without the use of an intermediate fluid, i.e., } } by using the critical point of water itself at 374 oC/3212 PSI? } } } } I'm interested in looking at clay samples. I am concerned that acetone will } } cause structural rearrangement of the clay particles because of dissimilar } } physical and electrical properties between acetone and water. Unlike } } biological specimens, clay would not care the least about a temperature of } } 374 degrees, as it does not begin its dehydroxylization process until over } } 500 degrees. I work in an oil field related industry where we have ready } } access to equipment that would consider 5000 PSI to be "light duty" so an } } appropriate pressure bomb would not be difficult to construct. } } } } Some questions: } } } } 1) All CPD devices that I have seen have a window that allows you to observe } } the state of the meniscus. Is this essential, or can I assume that the } } process will go as physics says it should without actually observing the } } meniscus? } } } } 2) I do not want to place the clay samples in water and allow the water to } } provide pressure as it is heated, as any water in contact with the clay will } } start to decompose the sample. Can I use an external pressure source (high } } pressure air, for example) to keep the pressure inside the bomb high enough } } to avoid evaporation of any water until the critical point is reached? Would } } it suffice to simply build a little platform that would keep the clay sample } } above the water level? } } } } 3) Am I nuts for even considering this? What am I missing? } } } } Thanks for your help. } } } } Bruce Girrell } } Microline Technology Corp. } } 2397 Traversefield Dr. } } Traverse City, MI 49686 } } http://www.microlinetc.com } } } } (231) 935-1585 (Voice) } } (231) 922-5099 (FAX) } } bigirrell-at-microlinetc.com } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Johathan, I have the same Sony camera match on my Axiophot and Axiovert . I have couple the cameras using optical couplers with a 0.6X magnification. This overcomes some of the magnification factor and allows a much larger section of the field to be captured. The couplers are made by Diagnostic Instruments. If you want more info let me know.
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Thursday, May 31, 2001 4:00:PM To: Microscopy-at-sparc5.microscopy.com
Hi:
A colleague wishes to get guidance and information about coupling his video camera to his compound microscope.
He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet lens mount. He wants to use it on a Zeiss Axiophot.
I am used to simple C-mount adapters, the bayonet mount is the ringer for me.
According to his research Sony offers an adapter, HRT045ENG12, that they say is used to mount this camera to a microscope. His question is 'Does the Sony adapter substitute for those offered by Zeiss for this purpose, or is the Sony adapter used to convert the bayonet mount to something compatable with the adapters offered by Zeiss?'
Finally, back in the old days, I remember learning that one could use a simple T-mount to attach a 35 mm camera to a microscope, but that it was not as good as having an eyepiece camera because it lacked the ocular lens. Is the same true of mounting a video camera? I have always just used simple, lenless C-mounts and it has seemed fine. If we are looking for ultimate image quality, should we be using a more sophisticated adapter?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Hi Jonathon, Remember that the image projected by the ocular of an LM on the CCD array is REAL ("Please draw a diagram of image formation in your light microscope", he asked the students in the audience!). The lens in the 'C' mounts are usually reduction lenses. [I haven't seen them all, so I am depending on my long-held view that if one wanted more resolution, one would use better objectives and 4x5" film(plates.] One may go without a reduction lens on camera and adapter, but then the video image will only be a subset [whose size (area) is dependent on the length of the 'C' adapter] of the area that one 'sees' in the 'virtual' image space. I have a Nikon bayonet-'C' adapter for a cooled CCD camera with a built-in reduction lens. My purpose was to extend the camera for some macro work using Nikon macro lenses that I already had for my Nikon 35mm camera. I also have two different types of 'C' mount adapter. The lensLESS adapter is used with a CCD camera that already has a reduction lens fitted to it, while the lensED adapter is used with a vidicon that lacks a reduction lens. I mismatched the adapters once and used the lensLESS adapter on the lensLESS vidicon (did use the ocular!). Only a subset, and no intensity! The only way to make a decision is to try both Nikon and Zeiss bayonet adapters, because one cannot determine - except empirically - what design criteria the engineers used, though my suspicion would be that they would not differ by much if both are used with an ocular. Also, watch the length of the adapter. A little long, and the camera (I am not familiar with its form.) may have to be connected more securely to avoid vibration effects.
READ ON AT YOUR OWN RISK (OF WASTING TIME!)
Film Photomicrography? How many listers have ever used a bellows camera setup for photomicrography? Your age is determined by the thickness of the catalog you received from the manufacturer of your microscope. What ARE we going to do with all that stuff in the darkroom?
Leitz ORTHOLUX? Catalogs? Speaking of that, I still have catalogs of Leitz (now Leica) from the OrthoLUX epoch (1950's-1970+). If anyone needs something, a picture or a number, don't hesitate to ask.
Video on TEM in 80's? I remember an old TEM that I once used had a video (vidicon) camera mounted under the viewing plate. The computer resided against half of a 9 foot wall, and was ROBUST. A user wanted to count mitochondria in the cytoplasm of cells. The only way I could get a single mitochondrion in the video camera view was to reduce the TEM mag almost to scan. So, while I could see a large part of the grid on the EM view screen the camera 'grabbed' only the center of a grid square and didn't show a bar! To program the computer, one had to remove boards and change jumpers. AH! The good OLD days!
Regards to all,
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Thursday, May 31, 2001 3:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM- Video camera/microscope coupler } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } } A colleague wishes to get guidance and information about coupling his } video } camera to his compound microscope. } } He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet } lens mount. He wants to use it on a Zeiss Axiophot. } } I am used to simple C-mount adapters, the bayonet mount is the ringer for } me. } } According to his research Sony offers an adapter, HRT045ENG12, that they } say is used to mount this camera to a microscope. His question is 'Does } the } Sony adapter substitute for those offered by Zeiss for this purpose, or is } the Sony adapter used to convert the bayonet mount to something compatable } with the adapters offered by Zeiss?' } } Finally, back in the old days, I remember learning that one could use a } simple T-mount to attach a 35 mm camera to a microscope, but that it was } not as good as having an eyepiece camera because it lacked the ocular } lens. } Is the same true of mounting a video camera? I have always just used } simple, lenless C-mounts and it has seemed fine. If we are looking for } ultimate image quality, should we be using a more sophisticated adapter? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
This reply is a bit off the Microscopy theme of the post, but here goes ---
Adding high pressure air to a "pressure bomb" will not prevent the evaporation of the water (though it will slow it down). The vapor pressure of any gas in equilibrium with its liquid phase is independent of the pressure of other gases present. In Bruce's example, if he added air to give a pressure of 3212psi at the critical temperature, the pressure in the container would actually rise (assuming enough water was present) to 6424psi. This is why, when using a pressure cooker, you have to allow the steam to vent for a few moments before closing the valve and allowing the pressure to rise - the boiling point of the water depends only upon the pressure of the water vapor above the liquid, not the total pressure.
Tony Garratt-Reed.
At 05:07 PM 5/31/2001 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
I don't have a LaB6 XL30, but I used to have its predecessor, a very early ElectroScan E3. We ran that with LaB6 in a routine vacuum of about 5 - 7x10-7 Torr. It did not have the capability of being baked. The LaB6 ran fine - we almost never were able to run filaments to their "natural" life, as the manual control allowed students to abuse the current and emission ratings - a facility they took advantage of regularly, destroying the filaments in the process! However, we certainly had filaments run for up to 1000 hrs.
As it aged (it was 10 years old when we replaced it, and still with its original ion pump) we did not experience ultimate vacuum changes, although the ion pump did become noticably harder to start after a gun vent.
As a general comment, I can't believe that a well-designed vacuum system (as I assume the thermionic XL30 is), pumped by an ion pump and baked, can't maintain a vacuum in the low -7's, unless there is a leak or some other problem.
Tony G-R
} -----Original Message----- } } From: FABBRI {fabbri-at-cigssrv1.unimo.it} } To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} } Date: Thursday, May 31, 2001 6:39 AM } Subject: XL30 - LaB6 Vacuum system } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
I'd like to echo the comments of Scott and Roger, who replied earlier. Their wisdom is worth repeating. I wrote SOPs for a commercial analytical service (not SEM specifically) in a past position........including the SOP on "writing SOPs". Meet the requirements, but do not go overboard with too many restrictions.
When considering a quality program it is so important to generate documentation that you can adhere to and follow. It is possible to create an incredibly "thorough" document which appears to cover every possible eventuality......but is also impossible to execute. Quality is then not achieved. Those who solicit feedback and allow their quality program to evolve over time will have more success. Having a strict internal review program that documents and corrects existing problems is actually a benefit in terms of passing external reviews. The internal documentation is typically the first thing examined by external examiners and it places you in a better light if it is clear you are making an effort to improve your program.
I would offer only a few suggestions for your specific analytical document:
If you have protocols for specific types of analyses that require more detailed instruction incorporate these as separate work instructions that are distinct from your SOP. The advantage is you can format your SOP in such a manner that the WIs can be amended or additional WIs can be added, without having to formerly generate an entire new version of your SOP. I have omitted the details, but consult one of the many manuals offered by the host of officials registrars. Referencing manuals is also fine for broader scope descriptions and procedures.
Maintanance Records and Statistical Process Control (SPC) procedures (i.e., calibration/sensitivity of EDS for example) are important components of your quality documentation.
A certain amount (not all) of the quality program information for commercial analytical service companies is available to customers (public). You may find some useful information by requesting information from their quality manager and/or asking specific departments or individuals about their quality procedures.
Very old book, perhaps on someone's shelf (Old instructor of EM class) Kay, D.H.,1965), Techniques for Electron Microscopy(2nd ed.), Chapter 8, "Preparation of Thin Sections" (Glauert, A.M. and Phillips, R.), Section: "Metallurgical and Crystallographic Applications of Thin Sectioning", pp. 248-253 (includes references), F.A. Davis, Philadelphia, PA, USA[Reprinted by Blackwell Scientific Pubs, Great Britain., 1967].
Kay, et al. promise that Zn is in there somewhere.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Dujin Wang } Sent: Thursday, May 31, 2001 3:54 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM need help on embedding and cutting of ZnO crsytals } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues, } I am trying to do ultrathin sectioning work of ZnO crystals, which size } is about 2-3microns(L) and 1-2microns(W). I am really a freshman } on TEM, so I hope to get help on this work. Any suggestions for the } choosing of embedding media and following procedures will be greatly } appreciated. } Thank you in advance. } } Dujin Wang } Max-Planck-Institute for Polymer Research } Ackermannweg 10, Mainz D-55128 } Germany } } Tel: 0049-6131-379226 } Fax: 0049-6131-379100 } Email: wangd-at-mpip-mainz.mpg.de } } }
Corn oil is used because it has lots of double bonds for the osmium to react with, so any highly unsaturated vegetable oil would do, or even be better than corn oil. Hey! Maybe there is a use for Vegemite ...
Phil
} Hello all, } Would anyone be able to tell me why corn oil is often recommended as the } correct oil to use to neutralise osmium tetroxide waste? Would any } vegetable oil do just as well, or does corn oil have special properties? } I have been unable to find it in supermarkets, and wonder if another } kind would be just as good. } Thanks. } Lyn Waterhouse } CEMMSA } Centre for Electron Microscopy } and Microstructure Analysis } University of Adelaide } Adelaide SA 5005 } Ph: (08) 8303 4074 } Fax: (08) 8303 4356 } Website: http://www.adelaide.edu.au/CEMMSA
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
This post was a few days ago - apologies for the delayed response, but I've not seen others make quite the same points.
SOP's are merely recipes for achieving a goal. In Colin's case, he is needing to achieve quality control, others may be needing safe operation, ISO standard certification, or whatever. The point is that the SOP will vary depending on the goal. There is no single "Standard Operating Procedure" that covers every operating mode (unless it is so full of qualifications, conditions, and alternatives as to be practically useless.) For EDX, the manufacturer's instructions may be a good guide, and at least a useful reference.
After stating the goal, the SOP will put down, in writing, all the essential steps that must be followed to achieve this goal. This will include such items as checking the performance of the system (mag. and beam voltage calibration of the microscope, energy calibration and resolution of the EDX, for example), as well as things like specimen preparation and mounting (for quant. EDX, the specimen must be polished, and mounted in an accurately known orientation, etc., etc.).
Hope this helps.
Tony Garratt-Reed
} -----Original Message----- } Sent: Tuesday, May 29, 2001 1:20 AM } To: MSA Listserver } Subject: Writing SOP's for SEM/X-Ray Analysis } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
You could freeze-dry specimens and that would be a rather more obvious approach. The cost of building a high temperature/ pressure bomb aside the question is: what would be the effect of the high temperature on the structure of the clay. I think what is missing from your consideration is that water is intrinsic to clay and its nature is changed irreversible when all water is removed from its chemical structure. I know that quite a few people have worked on the fine structure of clay and it appears that this seemingly simple task is completely elusive. I don't know, but I am doubtful that much useful could be gleaned from modest SEM magnifications of dried clay. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, June 01, 2001 7:07 AM, Bruce Girrell [SMTP:bigirrell-at-microlinetc.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Has anyone here familiar with any attempts to achieve critical point drying } of a water saturated sample without the use of an intermediate fluid, i.e., } by using the critical point of water itself at 374 oC/3212 PSI? } } I'm interested in looking at clay samples. I am concerned that acetone will } cause structural rearrangement of the clay particles because of dissimilar } physical and electrical properties between acetone and water. Unlike } biological specimens, clay would not care the least about a temperature of } 374 degrees, as it does not begin its dehydroxylization process until over } 500 degrees. I work in an oil field related industry where we have ready } access to equipment that would consider 5000 PSI to be "light duty" so an } appropriate pressure bomb would not be difficult to construct. } } Some questions: } } 1) All CPD devices that I have seen have a window that allows you to observe } the state of the meniscus. Is this essential, or can I assume that the } process will go as physics says it should without actually observing the } meniscus? } } 2) I do not want to place the clay samples in water and allow the water to } provide pressure as it is heated, as any water in contact with the clay will } start to decompose the sample. Can I use an external pressure source (high } pressure air, for example) to keep the pressure inside the bomb high enough } to avoid evaporation of any water until the critical point is reached? Would } it suffice to simply build a little platform that would keep the clay sample } above the water level? } } 3) Am I nuts for even considering this? What am I missing? } } Thanks for your help. } } Bruce Girrell } Microline Technology Corp. } 2397 Traversefield Dr. } Traverse City, MI 49686 } http://www.microlinetc.com } } (231) 935-1585 (Voice) } (231) 922-5099 (FAX) } bigirrell-at-microlinetc.com
Would you consider (freeze-fracturing, freeze-drying) or (freeze-drying, dry-fracturing) techniques? How about cryo-SEM and ESEM? If you decide to build and use a super CPD, please let us know the results.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } "Bruce Girrell" {bigirrell-at-microlinetc.com} 05/31 5:07 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Has anyone here familiar with any attempts to achieve critical point drying of a water saturated sample without the use of an intermediate fluid, i.e., by using the critical point of water itself at 374 oC/3212 PSI?
I'm interested in looking at clay samples. I am concerned that acetone will cause structural rearrangement of the clay particles because of dissimilar physical and electrical properties between acetone and water. Unlike biological specimens, clay would not care the least about a temperature of 374 degrees, as it does not begin its dehydroxylization process until over 500 degrees. I work in an oil field related industry where we have ready access to equipment that would consider 5000 PSI to be "light duty" so an appropriate pressure bomb would not be difficult to construct.
Some questions:
1) All CPD devices that I have seen have a window that allows you to observe the state of the meniscus. Is this essential, or can I assume that the process will go as physics says it should without actually observing the meniscus?
2) I do not want to place the clay samples in water and allow the water to provide pressure as it is heated, as any water in contact with the clay will start to decompose the sample. Can I use an external pressure source (high pressure air, for example) to keep the pressure inside the bomb high enough to avoid evaporation of any water until the critical point is reached? Would it suffice to simply build a little platform that would keep the clay sample above the water level?
3) Am I nuts for even considering this? What am I missing?
Thanks for your help.
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
} ****************************************************************************** } ************************************************* } } International SEMATECH, a consortium of worldwide semiconductor manufacturing } companies, has an opening for a Materials Analysis Laboratory Manager. We } offer our employees an attractive package of benefits, including medical and } dental coverage, group legal insurance, college tuition reimbursement, 12 paid } holidays plus vacation. Additionally, we offer a generous retirement and } shared savings plan, in which you are fully vested after two years of } employment, PLUS a bonus program. } } To learn more about International SEMATECH, visit our website at } www.sematech.org. To apply for the Materials Analysis Laboratory Manager } position, send your resume to: staffing.hr-at-sematech.org. Principals only } please. } } } ****************************************************************************** } ***************** } } Materials Analysis Laboratory Manager Job Description } Job Summary: } Management of the daily operation in the (Advanced Tool Development Facility) } ATDF's Materials Analysis Laboratory in a cost effective method, including all } logistical planning and scheduling activities. Continual interactions with the } customer on the prioritizing and dispositioning of data and samples is a major } component of this job. } Job Responsibilities: } * Maintain a safe working environment: be mindful of any potential } hazards, report and/or correct anything that needs to be made safe, } participate in monthly group self-inspections. Work with safety to update } policies and procedures and ensure they are appropriate. } * Maintain group specific safety training records. Manage technical } leadership of group to "realize the roadmap" } * Maintain a quality-staffing plan through guidance and feedback, } encourage appropriate training, professional interaction, presentations and } publications, retain (attract if needed) and develop quality staff. } * Maintains & Reports on the MA lab's budget to both management and } customers. } * Ownership of MA indices and develops plans for continuous improvement of } those indices } * Promotes team interactions to optimize resources with sister FA lab } * Provides the highest level of Customer satisfaction and Member Company } value. } * Maintain leading edge tool set through creative means, i.e. partnering, } reduced cost of purchase, etc. } Qualifications: } * Minimum 4 years experience in management of an analytical laboratory in } semiconductor industry } * Current and comprehensive working knowledge of semiconductor } manufacturing technology and associated characterization and metrology } equipment and techniques } * Direct semiconductor industry experience with all or most of the } following techniques: Auger, SIMS-ICP-MS, TEM, SEM, FIB/SIM, AFM, TXRF, and } VPD-ICP-MS } * Degree in: Material Sciences, Surface Chemistry, Physics or Chemical } Engineering preferred, PhD, MS, EE and/or BS with commensurate experience. } * Currently Published articles in related field(s) } Special Conditions: } * Travel Required {10% of the time. } * Shift/Hours worked:1st }
Lyn Waterhouse wrote: ================================================= Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. ============================================= Whether one kind of oil works better vs. another is a function of the unsaturation present. However, from an environmental and also a recycling standpoint, none of these "oils" is very environmentally friendly because once the osmium is put into this state, the economics of recovery become so terrible, that the concept of recovery and recycling becomes impossible (at least at current market prices). The only fate is incineration and burial in a land fill.
SPI Supplies is now beta testing our own SPI Osmium Recyling Kit. It consists of a 4 liter bottle with some formulated "potion" inside. When the bottle is filled up, the idea is for it to be returned to SPI Supplies for recycling. The chemistry is not rocket scientist logic, but navigating the various regulatory challenges, especially since they tend to vary from country to country, not to mention the rules being constantly changing as well, is what presents the real challenges. We believe the economics are such that we could even "give back" some new osmium tetroxide in amoules upon the return of such a bottle.
We would be happy to send you, with our compliments, a beta test kit. Just let us know if you would like us to do that. We would give you instructions for returning it to SPI for recycling. We are doing this on a carefully controlled basis for now, because every time we think we have all the bases covered, we encounter some other surprise. However, we plan to make some kind of more formal announcement about it on our website later this year, if not earlier at the M&M meeting, when the kit is officially introduced as a regular product of SPI Supplies.
But in the mean time, rather than doing the neutralization in an "oil", use 1N KOH solution. You will get the final reduction of the tetroxide to the dioxide, and the resulting system could at some future time, literally be "added" to the SPI Osmium Recycling Kit when it is more widely available. Now this suggestion might not be right for everyone but it is an alternative to rendering the material to a fate outside of the stream of commerce from which it is then lost for the benefit of future generations.
Disclaimer: SPI Supplies would like to promote the concept of recycling instead of incineration and burial of this non-renewable resource. This is called "passionate commercialism", something I hope is permitted on this listserver......!
Chuck
=============================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
What volume ratio of 0.1 M KOH to OsO4 (say, 1% in buffer) do you recommend?
Marie
You wrote:
} But in the mean time, rather than doing the neutralization in an "oil", use } 1N KOH solution.
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
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Hello all, Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. Thanks. Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
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This reply is a bit off the Microscopy theme of the post, but here goes ---
Adding high pressure air to a "pressure bomb" will not prevent the evaporation of the water (though it will slow it down). The vapor pressure of any gas in equilibrium with its liquid phase is independent of the pressure of other gases present. In Bruce's example, if he added air to give a pressure of 3212psi at the critical temperature, the pressure in the container would actually rise (assuming enough water was present) to 6424psi. This is why, when using a pressure cooker, you have to allow the steam to vent for a few moments before closing the valve and allowing the pressure to rise - the boiling point of the water depends only upon the pressure of the water vapor above the liquid, not the total pressure.
Tony Garratt-Reed.
At 05:07 PM 5/31/2001 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
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Dear XL30 LaB6 users
We have a LaB6 equipped ESEM installed in early 2000. It is still using the original filament which has now logged over 1000 hours. Although we have had various problems with the instrument vacuum levels have not been part of it. Gun vacuum is in the range 2.0 - 2.6 x10-7.
We bake it out overnight every time air is admitted to the gun for cleaning or repair. We are in the process of installing a high vac valve between the ion pump and the gun to save on bake outs at pressured times.
Sounds like you have a 'microleak' - get the agents onto it while you are still under warranty ! Baking out every two weeks might appeal to the purists who want to ensure 'ultimate vacuum' but it seems like overkill - unless it is compensating for a latent problem.
Another point that may be of interest - we have found that the instrument (with mixed lo and hivac usage) will last about three months before it starts getting wobbly ie gun instability and uncorrectable astigmatism. This is an indicator to clean the Wehnelt - itself no mean task since you are cleaning off an entirely transparent but totally nonconductive layer of Lanthanum (?) from the inside of the cap - checking for continuity with a multimeter as you clean. Once baked out after this process it performs like a star once again ! Thus far we are cleaning with polish, but are investigating less exhausting chemical cleaning methods !
The local service agents for FEI/Philips Anaspec-at-icon.co.za have assisted us a great deal in coming to terms with the idiosyncrasies of a LaB6 equipped ESEM - a concept which has given us some interesting moments.
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0) 33 260 5155 Fax +27 (0) 33 260 5776 website via:http://www.nu.ac.za/department/default.asp?dept=cemunp Email:bruton-at-nu.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
} } } "FABBRI" {fabbri-at-cigssrv1.unimo.it} 05/31/01 09:55AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
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Access to cryoSEM or ESEM would save a lot of work.
(Without any real experience of clays) I would be inclined to just air dry the sample at 40 deg. C in an oven.
The meniscus is required when replacing the intermediate fluid, to prevent exposing the sample, since you would not have one I guess it is irrelevant.
Dave
On Fri, 01 Jun 2001 10:06:22 +1000 John V Nailon {J.Nailon-at-mailbox.uq.edu.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } G'day Bruce, } What is wrong in going the other way and simply dry your sample using a } small vacuum chamber?? } High temperature and high pressure spell trouble to me. } Regards } JVN } } Bruce Girrell wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Has anyone here familiar with any attempts to achieve critical point drying } } of a water saturated sample without the use of an intermediate fluid, i.e., } } by using the critical point of water itself at 374 oC/3212 PSI? } } } } I'm interested in looking at clay samples. I am concerned that acetone will } } cause structural rearrangement of the clay particles because of dissimilar } } physical and electrical properties between acetone and water. Unlike } } biological specimens, clay would not care the least about a temperature of } } 374 degrees, as it does not begin its dehydroxylization process until over } } 500 degrees. I work in an oil field related industry where we have ready } } access to equipment that would consider 5000 PSI to be "light duty" so an } } appropriate pressure bomb would not be difficult to construct. } } } } Some questions: } } } } 1) All CPD devices that I have seen have a window that allows you to observe } } the state of the meniscus. Is this essential, or can I assume that the } } process will go as physics says it should without actually observing the } } meniscus? } } } } 2) I do not want to place the clay samples in water and allow the water to } } provide pressure as it is heated, as any water in contact with the clay will } } start to decompose the sample. Can I use an external pressure source (high } } pressure air, for example) to keep the pressure inside the bomb high enough } } to avoid evaporation of any water until the critical point is reached? Would } } it suffice to simply build a little platform that would keep the clay sample } } above the water level? } } } } 3) Am I nuts for even considering this? What am I missing? } } } } Thanks for your help. } } } } Bruce Girrell } } Microline Technology Corp. } } 2397 Traversefield Dr. } } Traverse City, MI 49686 } } http://www.microlinetc.com } } } } (231) 935-1585 (Voice) } } (231) 922-5099 (FAX) } } bigirrell-at-microlinetc.com } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Hi !!! I was doing EDAX on a sample to determine the surface constituents. Can you let me know answers to the following questions: 1. When I vary the accelerating voltage the atomic % (concentration of = } elements) varies. Why is that? 2. What is the depth of penetration of EDAX? Does this depend on = } accelerating voltage? Please send copies of your reply to bala-at-sc.edu. Thanks in advance Sincerely, Bala Haran
Hi !!! I was doing EDAX on a sample to determine the surface constituents. Can you let me know answers to the following questions: 1. When I vary the accelerating voltage the atomic % (concentration of = } elements) varies. Why is that? 2. What is the depth of penetration of EDAX? Does this depend on = } accelerating voltage? Please send copies of your reply to bala-at-sc.edu. Thanks in advance Sincerely, Bala Haran
Optem and Diagnostic Instrument make adapters for this camera to most any microscope.
gary g.
At 12:59 PM 5/31/2001, you wrote:
} Hi: } } A colleague wishes to get guidance and information about coupling his video } camera to his compound microscope. } } He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet } lens mount. He wants to use it on a Zeiss Axiophot. } } I am used to simple C-mount adapters, the bayonet mount is the ringer for me. } } According to his research Sony offers an adapter, HRT045ENG12, that they } say is used to mount this camera to a microscope. His question is 'Does the } Sony adapter substitute for those offered by Zeiss for this purpose, or is } the Sony adapter used to convert the bayonet mount to something compatable } with the adapters offered by Zeiss?' } } Finally, back in the old days, I remember learning that one could use a } simple T-mount to attach a 35 mm camera to a microscope, but that it was } not as good as having an eyepiece camera because it lacked the ocular lens. } Is the same true of mounting a video camera? I have always just used } simple, lenless C-mounts and it has seemed fine. If we are looking for } ultimate image quality, should we be using a more sophisticated adapter? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
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Hi Jonathon, Remember that the image projected by the ocular of an LM on the CCD array is REAL ("Please draw a diagram of image formation in your light microscope", he asked the students in the audience!). The lens in the 'C' mounts are usually reduction lenses. [I haven't seen them all, so I am depending on my long-held view that if one wanted more resolution, one would use better objectives and 4x5" film(plates.] One may go without a reduction lens on camera and adapter, but then the video image will only be a subset [whose size (area) is dependent on the length of the 'C' adapter] of the area that one 'sees' in the 'virtual' image space. I have a Nikon bayonet-'C' adapter for a cooled CCD camera with a built-in reduction lens. My purpose was to extend the camera for some macro work using Nikon macro lenses that I already had for my Nikon 35mm camera. I also have two different types of 'C' mount adapter. The lensLESS adapter is used with a CCD camera that already has a reduction lens fitted to it, while the lensED adapter is used with a vidicon that lacks a reduction lens. I mismatched the adapters once and used the lensLESS adapter on the lensLESS vidicon (did use the ocular!). Only a subset, and no intensity! The only way to make a decision is to try both Nikon and Zeiss bayonet adapters, because one cannot determine - except empirically - what design criteria the engineers used, though my suspicion would be that they would not differ by much if both are used with an ocular. Also, watch the length of the adapter. A little long, and the camera (I am not familiar with its form.) may have to be connected more securely to avoid vibration effects.
READ ON AT YOUR OWN RISK (OF WASTING TIME!)
Film Photomicrography? How many listers have ever used a bellows camera setup for photomicrography? Your age is determined by the thickness of the catalog you received from the manufacturer of your microscope. What ARE we going to do with all that stuff in the darkroom?
Leitz ORTHOLUX? Catalogs? Speaking of that, I still have catalogs of Leitz (now Leica) from the OrthoLUX epoch (1950's-1970+). If anyone needs something, a picture or a number, don't hesitate to ask.
Video on TEM in 80's? I remember an old TEM that I once used had a video (vidicon) camera mounted under the viewing plate. The computer resided against half of a 9 foot wall, and was ROBUST. A user wanted to count mitochondria in the cytoplasm of cells. The only way I could get a single mitochondrion in the video camera view was to reduce the TEM mag almost to scan. So, while I could see a large part of the grid on the EM view screen the camera 'grabbed' only the center of a grid square and didn't show a bar! To program the computer, one had to remove boards and change jumpers. AH! The good OLD days!
Regards to all,
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Thursday, May 31, 2001 3:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM- Video camera/microscope coupler } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } } A colleague wishes to get guidance and information about coupling his } video } camera to his compound microscope. } } He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet } lens mount. He wants to use it on a Zeiss Axiophot. } } I am used to simple C-mount adapters, the bayonet mount is the ringer for } me. } } According to his research Sony offers an adapter, HRT045ENG12, that they } say is used to mount this camera to a microscope. His question is 'Does } the } Sony adapter substitute for those offered by Zeiss for this purpose, or is } the Sony adapter used to convert the bayonet mount to something compatable } with the adapters offered by Zeiss?' } } Finally, back in the old days, I remember learning that one could use a } simple T-mount to attach a 35 mm camera to a microscope, but that it was } not as good as having an eyepiece camera because it lacked the ocular } lens. } Is the same true of mounting a video camera? I have always just used } simple, lenless C-mounts and it has seemed fine. If we are looking for } ultimate image quality, should we be using a more sophisticated adapter? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
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Hello all, Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. Thanks. Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
Staining cardiac vessel (e.g. aorta) wall is accompanied by a lot of autofluorescence, probably from elastin and perhaps other extracellualr matrix material.
Should attempts to reduce autofluorescence concentrate on the source of the problem, in other words, are there specific treatments for different types, such as lipofuscin versus elastin.
I have tried Sudan black B, sodium borohydride, toluidine blue. The first 2 reduced the red wavelength contribution only while toluidine blue actually increased the red autofluorescence. The green signal was reduced by only a small amount, all compared to just a PBS wash.
Any suggestions would be highly appreciated.
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital 30 Bond St. Toronto, ON M5B 1W8
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Access to cryoSEM or ESEM would save a lot of work.
(Without any real experience of clays) I would be inclined to just air dry the sample at 40 deg. C in an oven.
The meniscus is required when replacing the intermediate fluid, to prevent exposing the sample, since you would not have one I guess it is irrelevant.
Dave
On Fri, 01 Jun 2001 10:06:22 +1000 John V Nailon {J.Nailon-at-mailbox.uq.edu.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } G'day Bruce, } What is wrong in going the other way and simply dry your sample using a } small vacuum chamber?? } High temperature and high pressure spell trouble to me. } Regards } JVN } } Bruce Girrell wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Has anyone here familiar with any attempts to achieve critical point drying } } of a water saturated sample without the use of an intermediate fluid, i.e., } } by using the critical point of water itself at 374 oC/3212 PSI? } } } } I'm interested in looking at clay samples. I am concerned that acetone will } } cause structural rearrangement of the clay particles because of dissimilar } } physical and electrical properties between acetone and water. Unlike } } biological specimens, clay would not care the least about a temperature of } } 374 degrees, as it does not begin its dehydroxylization process until over } } 500 degrees. I work in an oil field related industry where we have ready } } access to equipment that would consider 5000 PSI to be "light duty" so an } } appropriate pressure bomb would not be difficult to construct. } } } } Some questions: } } } } 1) All CPD devices that I have seen have a window that allows you to observe } } the state of the meniscus. Is this essential, or can I assume that the } } process will go as physics says it should without actually observing the } } meniscus? } } } } 2) I do not want to place the clay samples in water and allow the water to } } provide pressure as it is heated, as any water in contact with the clay will } } start to decompose the sample. Can I use an external pressure source (high } } pressure air, for example) to keep the pressure inside the bomb high enough } } to avoid evaporation of any water until the critical point is reached? Would } } it suffice to simply build a little platform that would keep the clay sample } } above the water level? } } } } 3) Am I nuts for even considering this? What am I missing? } } } } Thanks for your help. } } } } Bruce Girrell } } Microline Technology Corp. } } 2397 Traversefield Dr. } } Traverse City, MI 49686 } } http://www.microlinetc.com } } } } (231) 935-1585 (Voice) } } (231) 922-5099 (FAX) } } bigirrell-at-microlinetc.com } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear Listers! Thank you very much for your interest in our problems. I got yours recommendation for HT check. Thank you very much again. Next week I will be in Moscow and I will be back on Friday June 8, 2001. Happy week end. Best regards Anton mailto:gut-at-thermo.isp.nsc.ru
Below is the result of your feedback form. It was submitted by (alsamszw-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, June 2, 2001 at 15:48:55 ---------------------------------------------------------------------------
Email: alsamszw-at-aol.com Name: Dr S Z AL-SAM
Organization: Mid Essex Hospitals, Chelmsford, UK
Education: Graduate College
Location: Chelmsford, United Kingdom
Question: Please tell me more about 3D microscopy and has this been used to study cytological smears before. Who supply these microscopes and how much do they cost? Many thanks.
Please tell me more about 3D images and 3D microscopes. Have these been used to study cytological smears and histological sections? How much do they cost and are there catalogues available? thank you AL-SAM
Research Technician (part-time) Electron Microscopy and Biochemistry.
We are seeking a technician preferably with experience in biochemistry and/or electron microscopy to join this BBSRC funded project “Trans-cellular Ca2+ transport and Ca2+ homeostasis in calcifying microalgae”. The post holder will be responsible for preparation and analysis of algal cells using electron microscopy in addition to purification of calcium binding proteins using gel electrophoresis.
The post is offered as a 50% fixed term appointment for 18 months with starting salary £13,717 (pro-rata), although other suitable working arrangements will be considered.
For further details of this project you can visit the following website: http://www.mba.ac.uk
Informal enquires may be addressed to Dr Alison Taylor (tel: +44 (0)1752 633290, e-mail: arta-at-mba.ac.uk) or Professor Colin Brownlee (tel: +44 (0)1752 633246, e-mail: cbr-at-wpo.nerc.ac.uk).
Closing Date: open until appointed but start by November 2001
Electron Microscopy(full time, permanent) Technician —Health Research Inc., Wadsworth Center, Albany, NY. We are looking for a motivated, mature and responsible individual to join a state-of-the-art Federal funded biomedical research laboratory. Flexible job hours in a challenging and stimulating research environment. Will train for specialized tasks.
Minimum qualification: B.S. in biology or related filed.
Preferred qualification: B.S. with experience in specimen sectioning and electron microscopy.
Responsibilities: 1. Embedding various biological specimens for transmission electron microscopy analyses. 2. Thin and thick (serially) sectioning of specimens. 3. Scanning and photographing sections with the EM 4. Conducting 3D analyses.
Contact: Richard Cole Research Scientist III Director of the Laser Microsurgery and Advanced Light Microscopy Core Unit Wadsworth Center P.O. Box 509 Albany N.Y. 12201-0509 rcole-at-wadsworth.org 518-474-7048 Phone 518-486-4901 Fax
We do alot of Immunofluorescence on sections of human skin. The main source of atuofluorescence is the elastin in the dermis and concentrated around the larger vessels. The best success we have had is using a CY5 label. The autofluorescence for elastin goes down dramaticly in the longer wavelengths. Or if we are using a single antiboby, we cature an image of the autofluorescence in a different channel and digitally subtract it out of the final image using image math. Then actually, most of the time we just leave it in and acknowledge that it is elastin (it is actually a good structural landmark and quite beautiful).
Bob Dermatology Research Center U of Washington, Seattle
On Fri, 1 Jun 2001, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Staining cardiac vessel (e.g. aorta) wall is accompanied by a lot of autofluorescence, probably from elastin and perhaps other extracellualr matrix material. } } Should attempts to reduce autofluorescence concentrate on the source of the problem, in other words, are there specific treatments for different types, such as lipofuscin versus elastin. } } I have tried Sudan black B, sodium borohydride, toluidine blue. The first 2 reduced the red wavelength contribution only while toluidine blue actually increased the red autofluorescence. The green signal was reduced by only a small amount, all compared to just a PBS wash. } } Any suggestions would be highly appreciated. } } } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital } 30 Bond St. } Toronto, ON M5B 1W8 } } ph: 416-864-6060 x6337 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } }
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
DIRECTOR, CELLULAR IMAGING CENTER: The Department of Pathology and Anatomy seeks Ph.D. applicants for a faculty position in fall 2001 as director of our cellular imaging core facility. The facility is equipped with transmission and scanning electron microscopes, a new laser scanning confocal microscope, image analysis computer, and conventional fluorescence microscopes.. Strong credentials in state of the art light and/or electron microscopy (e.g., confocal applications, FRET) are desired. In addition to overseeing the imaging core facility, the director will be expected to develop independent and/or collaborative research in one of five research areas: Cancer, Cardiovascular/Renal, Neuroscience, Reproductive Endocrinology, and Virology. Please send curriculum vitae, statement of research interests, and names, addresses, telephone numbers and email addresses of three references to:
William F. Glass II, M.D. Ph.D., Chair Department of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
Eastern Virginia Medical School is an Affirmative Action/Equal Opportunity Employer.
Questions may be directed to Dr. Glass or Dr. Earl Godfrey (godfreew-at-borg.evms.edu).
Please inform qualified individuals of this opportunity. Thank you very much.
Earl W. Godfrey, Ph.D. Dept. of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
street address: 700 W. Olney Rd. Lewis Hall Rm. 3077a Norfolk, VA 23507
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
DIRECTOR, CELLULAR IMAGING CENTER: The Department of Pathology and Anatomy seeks Ph.D. applicants for a faculty position in fall 2001 as director of our cellular imaging core facility. The facility is equipped with transmission and scanning electron microscopes, a new laser scanning confocal microscope, image analysis computer, and conventional fluorescence microscopes.. Strong credentials in state of the art light and/or electron microscopy (e.g., confocal applications, FRET) are desired. In addition to overseeing the imaging core facility, the director will be expected to develop independent and/or collaborative research in one of five research areas: Cancer, Cardiovascular/Renal, Neuroscience, Reproductive Endocrinology, and Virology. Please send curriculum vitae, statement of research interests, and names, addresses, telephone numbers and email addresses of three references to:
William F. Glass II, M.D. Ph.D., Chair Department of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
Eastern Virginia Medical School is an Affirmative Action/Equal Opportunity Employer.
Questions may be directed to Dr. Glass or Dr. Earl Godfrey (godfreew-at-borg.evms.edu).
Please inform qualified individuals of this opportunity. Thank you very much.
Earl W. Godfrey, Ph.D. Dept. of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
street address: 700 W. Olney Rd. Lewis Hall Rm. 3077a Norfolk, VA 23507
Below is the result of your feedback form. It was submitted by (davidbock-at-mindspring.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, June 3, 2001 at 20:06:47 ---------------------------------------------------------------------------
Email: davidbock-at-mindspring.com Name: David B.
Organization: --
Education: Graduate College
Location: LA, CA
Question: I would like to create a very large image of human blood at ~4000 times magnification. Ideally, it would be a single image that I would print at 10 feet x 30 feet. I imagine each red blood cell would be about 3" across in the final printed version.
Is it possible to take such a picture? Would I have to take a series of shots and then join them with Photoshop? What about resolution?
Below is the result of your feedback form. It was submitted by (bubidabub-at-worldnet.att.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 4, 2001 at 01:18:52 ---------------------------------------------------------------------------
Email: bubidabub-at-worldnet.att.net Name: Robin Olsen
Organization: Northampton Community College
Education: Undergraduate College
Location: Easton, PA Northampton County
Question: Could you please tell me what type of microorganisms one can expect to find in environmental dust taken from outdoor playground equipment? I am a college student taking microbiology at NCC and I am having a difficult time finding resources for research regarding organisms in dust and dirt. If you could tell me some things I could expect to culture from a sample taken from a playground, or tell me some resources where I could find this information I would greatly appreciate it! Thanks!!
Below is the result of your feedback form. It was submitted by (bubidabub-at-worldnet.att.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 4, 2001 at 01:18:52 ---------------------------------------------------------------------------
Email: bubidabub-at-worldnet.att.net Name: Robin Olsen
Organization: Northampton Community College
Education: Undergraduate College
Location: Easton, PA Northampton County
Question: Could you please tell me what type of microorganisms one can expect to find in environmental dust taken from outdoor playground equipment? I am a college student taking microbiology at NCC and I am having a difficult time finding resources for research regarding organisms in dust and dirt. If you could tell me some things I could expect to culture from a sample taken from a playground, or tell me some resources where I could find this information I would greatly appreciate it! Thanks!!
I have an old, but brilliant, book 'Leybold Vacuum Handbook' by K. Diels and R. Jaeckel (Translated by H. Adam and J. Edwards) Pergamon Press, 1966. It contains explainations of these problems, descriptions of experiments to measure outgassing and desorption, and rates for most elements and compounds that may be used in vacuum systems.
If you want a simple way to test the various materials find a pumping rig and pump it down to it's ultimate pressure at least overnight, ensure that the ultimate pressure is of the same order as your microscope. Then vent it to dry nitrogen and pump down again noting the pressure change with time to it's ultimate pressure (say for 1 hour). Then vent it and introduce the unknown component and pump down again noting the time and pressure. The change in pumping speed (rate of decrease in pressure) will be due to the outgassing. No fancy units but a direct comparison for your problem. Remember that to make a real comparison of outgassing rates you need to have similar amounts of material in similar forms, ie. same surface areas and volumes. However, I guess you are more concerned with the comparison between different mounting techniques so similar specimens mounted by the various techniques would be OK.
To save you a lot of time in experimenting I suggest that you use the minimum amount of mounting material, you want the smallest possible surface area to see the vacuum and you want the lowest desorption (outgassing) rate and lowest vapour pressure.
Of your 4 options I would not use sticky pads or waxes. I would only use epoxy or conductive inks and pastes fully underneath the specimen and ensure that the small amount used was fully cured and dried by pumping it out in a vacuum rig before loading it into the microscope. I would also store it under vacuum before loading.
Good luck, Ron
} } We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies: } } 1. sticky dots } 2. conductive inks and pastes } 3. epoxies } 4. waxes } } Is there an accepted method for rating these materials? } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Below is the result of your feedback form. It was submitted by (kellyburns-at-cfl.rr.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html#form on Monday, June 4, 2001 at 22:29:41 ---------------------------------------------------------------------------
Email: kellyburns-at-cfl.rr.com Name: Kelly Burns
Organization: Valencia Community College
Education: Undergraduate College
Location: Ocoee, FL, US
Question: When preparing a smear, why is it important to use tap water versus distilled water?
Karen Dye wrote the following: ========================================================== We just acquired a new high resolution SEM (FEG) and want to keep it "clean" . How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
Is there an accepted method for rating these materials? ========================================================== We at SPI Supplies have been dealing with this particular issue for more than thirty years. I don't think your question has the kind of absolute answer you might have been seeking, for the following reasons:
a) There are at least several different sources of "sticky dots", and they do not all behave the same. Some off-gas more than others and some are more sensitive to the beam than others. Also other factors such as age and storage conditions can effect the behavior of some of these adhesive materials in vacuum.
b) The conductive inks and pastes similarly represent different compositions, they are not all the same, and do not all come from the same place, but in addition to that, the "off-gassing characteristics" also depend on how thick of a layer has been applied and whether there is a "skin" that forms on the top, resulting in a lingering "wet" area underneath the skin. This effect can be even more problematic for a paste. The details of the formulation can influence the tendency of the composition to form such a skin, which acts as a barrier to diffusion.
c) The typical epoxy can be cured using more or less hardener, and it can be cured at room temperature or at higher temperatures. A room temperature cure with less rather than more hardener increases the chances for there being more unpolymerized material being present to off-gas and cause problems. The use of more hardener and some residence time at a higher temperature tends to minimize the off-gassing effect.
d) Waxes tend to be nonconductive and beam sensitive. Some have plasticizers that migrate to the surface and off-gas. We try to avoid waxes as a mounting medium for this kind of application in any kind of SEM. I have waxes like dental waxes in mind in the making of that comment.
We have adopted our own "test" for UHV compatibility. Simply put, it is a measure of the deterioration of vacuum when a test specimen is inserted into a UHV system. We found that the SPI double sided conductive carbon sheet material, when a 10 mm square was used, was inserted into the UHV system, there was no visible "meter deflection" on the LED display for the vacuum. Since the adhesive is the same as what is used in the SPI carbon tape and double sided discs, we would expect the same result would be found for them as well. We know that some similar appearing materials offered by others will perform as well on such a test. But not in all instances. We know this might not be the perfect test, but over the years it has been quite effective in terms of validating those materials that are better for this kind of application than others. Obviously, the worst performing material, if just a tiny bit is used, could end up "passing" the test, just as an inordinately large piece of the SPI double sided conductive sheets could flunk on such a test. So it is important to standardize on the area to be exposed to the UHV conditions if one is doing such a comparative test.
Having said all of that, we believe that the "dry adhesives" are far better than wet paints. But we also believe that the best laboratory practice would involve the use of the smallest amount of adhesive possible, and for that reason, we usually recommend the use of the sheets, which are cut out to the size desired for each sample, rather than the discs for which the smallest is still probably bigger than what is needed for many samples. Of course my other factors, such as personal preference and experience come into play so I am sure there could be selection based on factors other than purely that of off-gassing.
Disclaimer: SPI Supplies offers all four types of mountants for use for the mounting of SEM samples.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I provide a copy of the following URL to a site with rich detail on imaging for those of us in the mood to learn.
http://www.tasi.ac.uk/welcome.html
This site is TASI: "Technical Advisory Service for Images". Suggest further, to find tree of info, click on "FRAMEWORK"
Regards to all,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
I am posting this for a college, Lauri Wyner, in the Pathology Core. I have no idea how to section this stuff...!
Thanks! Maria Ericsson
My question is: I have a carbon coated porous tantalum matrix approximately 1 cm in circumference and 2 mm thick. This matrix has fixed cells adhered to the surface and throughout. I would like to embed this, make slides and stain for H&E to confirm the presences of cells as well as immunohistochemistry to characterize them. I am looking for suggestions on how I can section this matrix while maintaining its overall structure. Any information would be greatly appreciated.
Lauri Wyner DF/HCC Central Pathology Cores Coordinator Harvard Medical School G1-126, Goldenson Building 220 Longwood Avenue Boston, MA 02115 Tele: (617) 432-4947 Fas: (617) 432-6474 Lauri_wyner-at-hms.harvard.edu
This is a test for being able to send from our institution to the MSA List server. MS Outlook97 is annoying in being somewhat inscrutable for sending html unless you know how to find it's hidden secrets. This we have done. : ) ------------------------------------- Name: Charles Gilbert VOC: (704) 355-0604 Carolinas Medical Center FAX: (704) 355-8424 Dept of Pediatric Research digPager: (704) 355-4088 : 2058 PO Box 32861 Charlotte, NC 28232-2861
DISCLAIMER: {"The opinions are my own and not necessarily shared by my employer."}
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Karen Dye wrote the following: ========================================================== We just acquired a new high resolution SEM (FEG) and want to keep it "clean" How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
Is there an accepted method for rating these materials? ========================================================== We at SPI Supplies have been dealing with this particular issue for more than thirty years. I don't think your question has the kind of absolute answer you might have been seeking, for the following reasons:
a) There are at least several different sources of "sticky dots", and they do not all behave the same. Some off-gas more than others and some are more sensitive to the beam than others. Also other factors such as age and storage conditions can effect the behavior of some of these adhesive materials in vacuum.
b) The conductive inks and pastes similarly represent different compositions, they are not all the same, and do not all come from the same place, but in addition to that, the "off-gassing characteristics" also depend on how thick of a layer has been applied and whether there is a "skin" that forms on the top, resulting in a lingering "wet" area underneath the skin. This effect can be even more problematic for a paste. The details of the formulation can influence the tendency of the composition to form such a skin, which acts as a barrier to diffusion.
c) The typical epoxy can be cured using more or less hardener, and it can be cured at room temperature or at higher temperatures. A room temperature cure with less rather than more hardener increases the chances for there being more unpolymerized material being present to off-gas and cause problems. The use of more hardener and some residence time at a higher temperature tends to minimize the off-gassing effect.
d) Waxes tend to be nonconductive and beam sensitive. Some have plasticizers that migrate to the surface and off-gas. We try to avoid waxes as a mounting medium for this kind of application in any kind of SEM. I have waxes like dental waxes in mind in the making of that comment.
We have adopted our own "test" for UHV compatibility. Simply put, it is a measure of the deterioration of vacuum when a test specimen is inserted into a UHV system. We found that the SPI double sided conductive carbon sheet material, when a 10 mm square was used, was inserted into the UHV system, there was no visible "meter deflection" on the LED display for the vacuum. Since the adhesive is the same as what is used in the SPI carbon tape and double sided discs, we would expect the same result would be found for them as well. We know that some similar appearing materials offered by others will perform as well on such a test. But not in all instances. We know this might not be the perfect test, but over the years it has been quite effective in terms of validating those materials that are better for this kind of application than others. Obviously, the worst performing material, if just a tiny bit is used, could end up "passing" the test, just as an inordinately large piece of the SPI double sided conductive sheets could flunk on such a test. So it is important to standardize on the area to be exposed to the UHV conditions if one is doing such a comparative test.
Having said all of that, we believe that the "dry adhesives" are far better than wet paints. But we also believe that the best laboratory practice would involve the use of the smallest amount of adhesive possible, and for that reason, we usually recommend the use of the sheets, which are cut out to the size desired for each sample, rather than the discs for which the smallest is still probably bigger than what is needed for many samples. Of course my other factors, such as personal preference and experience come into play so I am sure there could be selection based on factors other than purely that of off-gassing.
Disclaimer: SPI Supplies offers all four types of mountants for use for the mounting of SEM samples.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656
e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Listers, Can anyone refer me to a company or lab that can do sub-micron particle size and distribution studies, especially but not necessarily, in the southeast US? It involves a study injecting small wear debris particles into a skeletal joint. Thanks for any help. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor June 4, 2001 2:44 PM Cannon Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 (Ship to: 1000 Blythe Blvd ) Fax: 704-355-0589 Charlotte, NC 28232-2861 (Ship to: 28203 ) Winston.Wiggins-at-CarolinasHealthCare.org {mailto:WWiggins-at-Carolinas.org} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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The general question of outgassing and its effect on the performance of vacuum systems is discussed in some detail in Section 2.9 of my book, 'Vacuum Methods in Electron Microscopy' (see: http://www.2spi.com/catalog/books/book48.html and http://pup.princeton.edu/titles/6484.html for details).
There is one interesting way in which unexpectedly large amounts of solvents and gases can get introduced into an SEM when carbon or silver paint is used to mount a specimen. This can occur if the paint is smeared on a mounting stub in such a way that the entire area underneath the specimen is covered, and then only a relatively short time is allowed for the paint to dry. Then only the paint around the edges of the specimen will dry, while that underneath the specimen can remain wet. When inserted into the SEM the solvent from the wet paint underneath the specimen can slowly diffuse outward through the 'dry' paint around the edges, thereby providing a strong source of outgassing for a long time.
If you really must use a paing to mount specimens, I think it is best to only put a few small dabs of it at several spots around the periphery of the specimen. Then there will be a better opportunity for the solvent to evaporate out from each small dab, and if sufficient time is allowed for the drying process, no wet paint will be left trapped underneath the specimen.
Better yet, make some simple clips or clamps that will hold the specimen in place mechanically, without the use of solvents or adhesives. With a little ingenuity and some pliers you can fashion a clip to hold almost any specimen from a piece of heavy copper wire or a paperclip. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
My Oxford EDS x-ray detector needs to be returned to Oxford for repair, and unfortunately, I cannot locate the original cover for the EDS port on the microscope. Neither Oxford not JEOL can supply a cover. I do not want to lose the use of the SEM for imaging while the detector is repaired. The microscope is a JEOL JXA-840. As far as I know, the column is identical to the JSM-840. Is there anyone out there able and willing to help me with a sale, rent, or loan of a suitable cover until my x-ray detector is repaired.
Thank you.
Dave Wilbur
-- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University voice: 617-627-2163 Fax: 617-627-3443 email: dwilbu01-at-tufts.edu __________________________________
I can agree with Ron about the waxes. I've used a JEOL 6600F and Hitachi S-4700s and neither of these instruments likes wax. The wax will outgas so badly that the loadlocks won't pump down, so you never get it into the main chamber. Superglue-type glues work good (fully cured) and I've had no trouble with black carbon double-sided tape. I also use carbon (graphite) paint after a 15-minute bake at about 40-50 degrees C. A vacuum dryer could also be used if your specimen was heat-sensitive. I don't much care for silver paint; it takes too long to dry and is hard to remove if necessary. For the ultimate in cleanliness, remember to handle your mounts, stages, etc. (anything that goes into the main chamber) with gloves. This will prevent finger oils from being dispersed in the system.
Ron Doole wrote: ----------- {snip} --------------
} To save you a lot of time in experimenting I suggest that } you use the minimum amount of mounting material, you want } the smallest possible surface area to see the vacuum and } you want the lowest desorption (outgassing) rate and lowest } vapour pressure. } } Of your 4 options I would not use sticky pads or waxes. I } would only use epoxy or conductive inks and pastes fully } underneath the specimen and ensure that the small amount } used was fully cured and dried by pumping it out in a } vacuum rig before loading it into the microscope. I would } also store it under vacuum before loading. } } Good luck, } Ron } } } } } We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies: } } } } 1. sticky dots } } 2. conductive inks and pastes } } 3. epoxies } } 4. waxes } } } } Is there an accepted method for rating these materials? } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Position available in an independent testing lab in a suburb of Detroit, MI. Knowledge of ISI SS 40 required. Materials testing experience a plus. For information contact 734-668-3309 and leave message.
Several questions about safety.. What do you use for gloves when handling both resins and fixatives? What procedures are out there for precipitating lead? Is anyone aware of any suspected instances of EM chemical exposure (resins, osmium), documented cases and/or references about such? Our institution would appreciate any information to help them answer questions about this unique part of the lab.
Rons advice is all very sound, but you have overlooked the effect of temperature. Even a modest rise in temperature increases vapour pressure and therefore outgassing markedly. Unless a cold-trap is used the objective area is rather warmer than room temperature. To cure samples so that later outgassing is lessened, store specimens at a suitably increased temperature and under a modest vacuum. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, June 05, 2001 6:13 PM, Ron Doole [SMTP:ron.doole-at-materials.oxford.ac.uk] wrote: } } } HI Karen, } } I have an old, but brilliant, book 'Leybold Vacuum } Handbook' by K. Diels and R. Jaeckel (Translated by H. Adam } and J. Edwards) Pergamon Press, 1966. It contains } explainations of these problems, descriptions of } experiments to measure outgassing and desorption, and rates } for most elements and compounds that may be used in vacuum } systems. } } If you want a simple way to test the various materials find } a pumping rig and pump it down to it's ultimate pressure at } least overnight, ensure that the ultimate pressure is of } the same order as your microscope. Then vent it to dry } nitrogen and pump down again noting the pressure change } with time to it's ultimate pressure (say for 1 hour). Then } vent it and introduce the unknown component and pump down } again noting the time and pressure. The change in pumping } speed (rate of decrease in pressure) will be due to the } outgassing. No fancy units but a direct comparison for } your problem. Remember that to make a real comparison of } outgassing rates you need to have similar amounts of } material in similar forms, ie. same surface areas and } volumes. However, I guess you are more concerned with the } comparison between different mounting techniques so similar } specimens mounted by the various techniques would be OK. } } To save you a lot of time in experimenting I suggest that } you use the minimum amount of mounting material, you want } the smallest possible surface area to see the vacuum and } you want the lowest desorption (outgassing) rate and lowest } vapour pressure. } } Of your 4 options I would not use sticky pads or waxes. I } would only use epoxy or conductive inks and pastes fully } underneath the specimen and ensure that the small amount } used was fully cured and dried by pumping it out in a } vacuum rig before loading it into the microscope. I would } also store it under vacuum before loading. } } Good luck, } Ron } } } } } We just acquired a new high resolution SEM (FEG) and want to keep it } } "clean". How can I find out about out-gassing rates and the relative } } "cleanliness" of various sample preparation supplies: } } } } 1. sticky dots } } 2. conductive inks and pastes } } 3. epoxies } } 4. waxes } } } } Is there an accepted method for rating these materials? } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk
Dave, Unless you have been running an exceptionally clean and good vacuum, a rubber stopper of the correct size will do nicely as a temporary substitute. In particular, make sure that it's not too small so that it doesn't get sucked through the port at 15 psi. Otherwise, after a couple of hours of pumping, the outgassing should be minimal (better after overnight).
Clean the sides of the stopper with IPA or some similar solvent and use a very small amount of your favorite vacuum grease around the sides.
Ken Converse owner Quality Images third party SEM service Delta, PA
David Wilbur wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } and unfortunately, I cannot locate the original cover for the EDS port } on the microscope. Neither Oxford not JEOL can supply a cover. I do } not want to lose the use of the SEM for imaging while the detector is } repaired. The microscope is a JEOL JXA-840. As far as I know, the } column is identical to the JSM-840. Is there anyone out there able and } willing to help me with a sale, rent, or loan of a suitable cover until } my x-ray detector is repaired. } } Thank you. } } Dave Wilbur } } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } voice: 617-627-2163 } Fax: 617-627-3443 } email: dwilbu01-at-tufts.edu {mailto:dwilbu01-at-tufts.edu} } __________________________________ } } } } }
We currently use either Denka or Fei filaments in our FESEM which have generally given similar results over similar lifetimes . However for various reasons we are mow experiencing occasional stray magnetic field interference with corresponding deterioration in image quality . I have never really thought it through but what's actually happening at the tip ?..
1. Apart from occasional irritating beam 'sway' is physical damage being done to the tungsten tip with this interference and
2. Assuming a fluid zirconia film/ball exists at the tip of the tungsten filament is this being consumed more quickly influencing lifetime ?
Best regards
Martyn
From M Harris Device Engineering ESM Ltd , Cardiff Rd . Newport , South Wales NP10 8YJ .
A comprehensive microscopy salary survey was published in the old EMSA Bulletin in the early 80's. It is too old to be useful. More recently, there was a smaller survey in "Microscopy Today." Check with Don Grimes at: microtoday-at-mindspring.com
Perhaps it's time for a new comprehensive survey.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
"Rajesh Patel" {rpatel-at-umdnj.edu} on 06/05/2001 01:27:32 PM
To: {microscopy-at-sparc5.microscopy.com} cc:
I beleive that some time a go a salray survey was done for electron microscopy field. How and where can I access that info.
Rajesh Patel Robert Wood Johnson Medical School Dept. of Pathology Electron Microscopy Lab 675 Hoes Lane Piscataway, NJ 08854
Dear Dave, While the exact fit would be nice, the dimensions of the requirement would be nicer. Also, if you haven't called the JEOL service center near you, you might try that. I have parts galore, but I have to know more about the item.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: David Wilbur } Sent: Tuesday, June 5, 2001 3:37 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM/EDS Need port cover for JEOL 840 microscope } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } and unfortunately, I cannot locate the original cover for the EDS port } on the microscope. Neither Oxford not JEOL can supply a cover. I do } not want to lose the use of the SEM for imaging while the detector is } repaired. The microscope is a JEOL JXA-840. As far as I know, the } column is identical to the JSM-840. Is there anyone out there able and } willing to help me with a sale, rent, or loan of a suitable cover until } my x-ray detector is repaired. } } Thank you. } } Dave Wilbur } } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } voice: 617-627-2163 } Fax: 617-627-3443 } email: dwilbu01-at-tufts.edu } __________________________________ } } } }
I would suggest glycol methacrylate embedment (as a start) and thin sectioning with a carbide knife (expensive, but well worth the cost when materials are 'hard')
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Maria Ericsson } Sent: Tuesday, June 5, 2001 11:53 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: sectioning tantalum matrix } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am posting this for a college, Lauri Wyner, in the Pathology Core. } I have no idea how to section this stuff...! } } Thanks! } Maria Ericsson } } My question is: } I have a carbon coated porous tantalum matrix approximately 1 cm in } circumference and 2 mm thick. This matrix has fixed cells adhered to the } surface and throughout. I would like to embed this, make slides and stain } for H&E to confirm the presences of cells as well as immunohistochemistry } to characterize them. I am looking for suggestions on how I can section } this matrix while maintaining its overall structure. Any information would } } be greatly appreciated. } } } } Lauri Wyner } DF/HCC Central Pathology Cores Coordinator } Harvard Medical School } G1-126, Goldenson Building } 220 Longwood Avenue } Boston, MA 02115 } Tele: (617) 432-4947 } Fas: (617) 432-6474 } Lauri_wyner-at-hms.harvard.edu } } }
I keep running into users who believe that High magnification for an SEM, say 20,000X and greater is a benefit to EDS analysis. I have always been under the impression that high magnification is not a benefit to EDS due to the large interaction volume of X-rays. Comments please.
The keyword here is analysis. You are absolutely correct for this. The interaction volume will definitely be much larger than the particular feature that the beam can fully resolve. I preached this for many years and still do to a certain extent. However, it was shown to me by a colleague here after we had been arguing about it that in fact you can get meaningful qualitative information from areas much smaller than the interaction volume. He showed me that putting the beam on a surface feature on a thin film on glass that had a lateral spatial dimension less than 0.1 um and comparing it to an area adjacent to this area gave a significant difference in the spectra. Enough of a difference that it enabled him to pinpoint the problem. We do quite a lot of qualitative analysis and very little quantitative analysis. From that standpoint, higher magnifications can be quite useful, but not for quantitative analysis.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ingram, Mike [mailto:MIngram-at-rodel.com] Sent: Wednesday, June 06, 2001 11:06 AM To: 'Microscopy-at-MSA.Microscopy.Com'
I keep running into users who believe that High magnification for an SEM, say 20,000X and greater is a benefit to EDS analysis. I have always been under the impression that high magnification is not a benefit to EDS due to the large interaction volume of X-rays. Comments please.
I agree that good vacuum practice is the first line of defense in keeping your new FEG SEM clean. The use of most of these materials may dirty your microscope.
XEI Scientific does offer a system to remove outgassed hydrocarbons from the walls and atmosphere of your microscope. The details about our EVACTRON SEM-CLEAN system may be found at www.SEMCLEAN.com This system was specifically developed for keeping FEG SEMs clean.
Ron Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 (650)-369-0133
-----Original Message----- } From: Karen Dye {karen.dye-at-medtronic.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
} My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } and unfortunately, I cannot locate the original cover for the EDS port } on the microscope. Neither Oxford not JEOL can supply a cover. I do } not want to lose the use of the SEM for imaging while the detector is } repaired. The microscope is a JEOL JXA-840. As far as I know, the } column is identical to the JSM-840. Is there anyone out there able and } willing to help me with a sale, rent, or loan of a suitable cover until } my x-ray detector is repaired. } David,
There are actually at least two different ports that could be used for mounting of the xray detector or, in this case, the blanking plate for the removed detector.
Is it the high take off angle port or the side mount, circular port?
P.S. I would advise against using a rubber stopper. This technique could leave you needing more than a blanking plate.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 (480) 967-3946 "www.emlabservices.com"
I would like to hear from users out there about experience with the Sutter Lambda DG4 fluorescence source/filter changer. It looks very nice on paper. How is brightness/field uniformity/flexibility for general use. We want ratio capability which it appears good at, but we also want versatility for normal single fluor imaging at various wavelengths. Any comments about using this as opposed to a standard mercury, standard xenon, or monochromator for a general fluorescence microscopy facility are appreciated. Thanks- Dave --
************************************************************ Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Generally, nothing is happening to the tip by the magnetic fields as the only components that affect filament life are: pressure, overdriving, & poor filament construction.
Earl
----- Original Message ----- } From: {"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 06, 2001 7:19 AM
A rubber stopper, well-greased with appropriate grease, worked fine on my 840 in a similar situation, no degradation of vacuum. Just make absolutely sure that it's big enough that there's no chance of its being sucked right in, and that nobody knocks into it!
cheers
rtch
} } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } } and unfortunately, I cannot locate the original cover for the EDS port } } on the microscope. Neither Oxford not JEOL can supply a cover. I do } } not want to lose the use of the SEM for imaging while the detector is } } repaired. The microscope is a JEOL JXA-840. As far as I know, the } } column is identical to the JSM-840. Is there anyone out there able and } } willing to help me with a sale, rent, or loan of a suitable cover until } } my x-ray detector is repaired. } } } David, } } There are actually at least two different ports that could } be used for mounting of the xray detector or, in this case, } the blanking plate for the removed detector. } } Is it the high take off angle port or the side mount, } circular port? } } P.S. I would advise against using a rubber stopper. This } technique could leave you needing more than a blanking plate. Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I recently performed an x-ray map and line scan on a cross sectioned Ag on Cu sample at 20,000X (15kV) and was able to show little to no migration of the Ag into the Cu. I was very surprised at how sharp the demarcations between the layers were at 20,000 in the x-ray maps and line scans. I would be happy to share the data w/ any interested parties. It is my opinion that high magnification EDS analysis can be useful. Of' course one can lower the kV or work with very thin samples to push the limits.
Ric
-----Original Message----- } From: Ingram, Mike [mailto:MIngram-at-rodel.com] Sent: Wednesday, June 06, 2001 11:06 AM To: 'Microscopy-at-MSA.Microscopy.Com'
I keep running into users who believe that High magnification for an SEM, say 20,000X and greater is a benefit to EDS analysis. I have always been under the impression that high magnification is not a benefit to EDS due to the large interaction volume of X-rays. Comments please.
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EMF does not generally affect SEMs in the electron column, sufficient shielding being provided by the normal design of these areas. Once the beam leaves the final pole piece and enters the sample chamber, though, it is a different matter. You can tell if the major influence is in the chamber by doing a simple qualitative measure of the interference at different working distances. If the problem is greater at the longer working distance, then the EMF is influencing the beam during its traverse through the sample chamber.
The same is basically true for vibrational effects. Within the column, physical movement of the instrument laterally is somewhat cancelled by the changing proximity to the applied fields of the electron optic system. Once free of those fields, though, movement of the instrument (and thus the sample) change the apparent position of the beam. Once again, the effect is reduced with a reduction in the working distance.
The first step has to be to determine whether the problem is EMF or vibration - quite possibly both. Since many vibrational sources are linked to the electrical line frequency, this can sometimes be tough. However, most modern SEMs should be quite capable of damping 60Hz. vibrations, but if EMF of any low frequency can come in then all low frequencies will be allowed in - EMF shielding is far less specific than vibrational.
This leads to the usual trick of trying to determine the offensive frequency by counting the number of 'saw-tooth' edges in a portion of a slow image sweep. If the frequency is close to 60Hz then it is probably EMF.
If the appearance of a problem like this is rather sudden, then the natural tendency is to find a temporal link to some change in the system. Taking this approach, though, you have to be sure to include all possible changes that may have taken place. For both problems, you have to consider if any changes have been made in the environment around the SEM. Has any equipment been added or moved in the area that may affect the instrument? Can the effect be due to a seasonal change in the HVAC equipment in use? Have there been any increases in the current load on any power distribution lines near the instrument?
More intimate causes also have to be accessed. Have any electrical cables, air, water or vacuum lines on the SEM been repositioned? EMF isolation generally involves putting a distance between any sources and any receptors. Vibrational isolation generally involves that as well as several levels of damping that require isolation between them. A simple cable or tube that is moved and bridges the isolation of one or more stages can destroy their effectiveness. Moving a computer monitor or external vacuum gauge controller a few inches can make profound changes in received EMF.
Finally, one has to consider time. Time, along with other influences, can produce changes such as corrosion of electrical connections that can cause high resistance in ground connections (in this case a 'high' resistance of only a few ohms can cause ground loop currents in instruments that can become a real problem). Another problem with aging instruments is in the filter capacitors of their power supplies. Capacitors age, and in doing so, their values often change. If a circuit is very tightly designed, this change can result in an increase in a 60 Hz ripple seen in power supplies. If circuit designs rely heavily on bypass capacitors to reduce ripple to individual circuits from power supply circuits, a failure of one or more of those capacitors can also result in increased supply ripple to circuits. Such failures will have the appearance of an induced EMF effect, but without the dependence on working distance.
Seems like I've said a lot, but I haven't even scratched the surface. SEMs, and particularly FESEMs are extremely sensitive instruments. Their design, implementation and continued operation are a continual balance of many factors. However, the problems you describe are probably not related to the use of third-party emitters.
You didn't mention the make and model of the instrument you are using, or the particular orientation of the emitters you are using. Could you elaborate on these?
On Wednesday, June 06, 2001 9:19 AM, "HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com [SMTP:"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Dear Colleagues , } } We currently use either Denka or Fei filaments in our } FESEM which have generally given similar results over similar } lifetimes . } However for various reasons we are mow experiencing occasional stray } magnetic field interference with corresponding deterioration in image } quality . } I have never really thought it through but what's actually happening } at the tip ?.. } } 1. Apart from occasional irritating beam 'sway' is physical damage } being done to the tungsten tip with this interference and } } 2. Assuming a fluid zirconia film/ball exists at the tip of the } tungsten filament is this being consumed more quickly influencing } lifetime ? } } Best regards } } Martyn } } } From M Harris } Device Engineering } ESM Ltd , Cardiff Rd . } Newport , South Wales } NP10 8YJ . } } Email harrism-at-esm-semi.co.uk } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Dear all, The TNT (Trends in Nanotechnology) 2001 registration deadline at the special price of 425 Euros is approaching on June 15th. After this date, the registration fee will be 500 Euros. The conference is taking place in the first week of September this year in Segovia, Spain, bringing together the top names in Nanotechnology for a week's intensive brainstorming. The line up of keynote speakers includes representatives from NASA, IMEC, Texas Instruments, Sony & Samsung, as well as universities and research institutes in Europe, the US, & Japan.
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Full details can be found at http://www.cmp-cientifica.com/tnt2001 Regards Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
Interesting question. I generally let users know that the interactions of electrons and x-ray generation and flourescence limit the analytical volume to a sphere of 50 - 100 microns in diameter. However, that assumes that you want to quantify that specific volume. There are many times when you want different criteria. For example, if one were to do a line scan across the interface of two differing materials. In this case, we can artificially determine the boundary as a function of the constantly varying intensities of their different components even though the compositional change takes place in an area much smaller than we can reasonable measure.
Such measurement, however, has to be made with the caviat of the experimental conditions and the 'thresholds' used for determination. Ideally, these are determined using similar matrix standards - i.e., similar materials whose dimensional composition can be verified by other means.
Thin film or small particle determinations are made using assumptions that are probably quite accurate, but have to be taken with consideration of the experimental conditions and their effects on known compositions. One has to be careful to ensure that the assumptions made by a particular routine are applicable to their particular circumstances.
Case in point - you currently have the beam located on the exact center of the interface of two widely different materials. In this case, you would expect an equal contribution from both materials. However, you have to consider many factors of x-ray generation by incident electron radiation, the flourescence and cross-flourescence of the individual materials as well as the absorption of each for varying wavelengths. Now add into the mix the morpholgy of your specific samples. We don't currently have all the answers to these complex problems and rely, instead, on empirical data. The closer the empirical data to the materials you're looking at, the better the analysis.
In brief, very minor differences between extremely close materials is possible, but what you can make of those differences depends on the effort you put into it.
On Wednesday, June 06, 2001 10:06 AM, Ingram, Mike [SMTP:MIngram-at-rodel.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I keep running into users who believe that High magnification for an SEM, } say 20,000X and greater is a benefit to EDS analysis. I have always been } under the impression that high magnification is not a benefit to EDS due } to the large interaction volume of X-rays. Comments please. } } } Mike Ingram } Rodel Inc. }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Tantalum is very hard and tough, so I doubt that it will section with either diamond or tungsten carbide knives. Preparing the material like a geological section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. Cheers Jim Darley Disclaimer: ProSciTech supplies both diamond and TC knives. I advised against the use of either in this particular case. ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, June 06, 2001 1:54 AM, Maria Ericsson [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core. } I have no idea how to section this stuff...! } } Thanks! } Maria Ericsson } } My question is: } I have a carbon coated porous tantalum matrix approximately 1 cm in } circumference and 2 mm thick. This matrix has fixed cells adhered to the } surface and throughout. I would like to embed this, make slides and stain } for H&E to confirm the presences of cells as well as immunohistochemistry } to characterize them. I am looking for suggestions on how I can section } this matrix while maintaining its overall structure. Any information would } be greatly appreciated. } } } } Lauri Wyner } DF/HCC Central Pathology Cores Coordinator } Harvard Medical School } G1-126, Goldenson Building } 220 Longwood Avenue } Boston, MA 02115 } Tele: (617) 432-4947 } Fas: (617) 432-6474 } Lauri_wyner-at-hms.harvard.edu } }
we are in the process of cost our EM service, that is a cost breakdown for doing EM. i know the actual costs are very small. i was wondering if anyone has done it recently. we are a diagnostic lab. john hoffpauir cooper hospital
Just out of curiousity. . . how much X-radiation penetrates the rubber stopper?
Marie
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Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
I vote with Ritchie. Rubber stoppers are great for temporary problems, particularly for finding vacuum leaks. One stopper can eliminate a more complex assembly. I somehow prefer the silicone rubber stoppers. However, do consider X-ray penetration. Particularly on instruments above 20kV. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, June 07, 2001 10:44 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote:
} } } A rubber stopper, well-greased with appropriate grease, worked fine } on my 840 in a similar situation, no degradation of vacuum. } Just make absolutely sure that it's big enough that there's no chance } of its being sucked right in, and that nobody knocks into it! } } cheers } } rtch } } } } } } } } } } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } } } and unfortunately, I cannot locate the original cover for the EDS port } } } on the microscope. Neither Oxford not JEOL can supply a cover. I do } } } not want to lose the use of the SEM for imaging while the detector is } } } repaired. The microscope is a JEOL JXA-840. As far as I know, the } } } column is identical to the JSM-840. Is there anyone out there able and } } } willing to help me with a sale, rent, or loan of a suitable cover until } } } my x-ray detector is repaired. } } } } } David, } } } } There are actually at least two different ports that could } } be used for mounting of the xray detector or, in this case, } } the blanking plate for the removed detector. } } } } Is it the high take off angle port or the side mount, } } circular port? } } } } P.S. I would advise against using a rubber stopper. This } } technique could leave you needing more than a blanking plate. } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
In order to re-send this email, I had to change the subject line to stop a rejection because of the r word.
-----Original Message----- } From: Beauregard, Paul A. Sent: Thursday, June 07, 2001 11:39 AM To: Microscopy-at-sparc5.microscopy.com
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Jim Darley wrote:
==================================================== Tantalum is very hard and tough, so I doubt that it will section with either
diamond or tungsten carbide knives. Preparing the material like a geological
section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and
view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. ===================================================== Actually Ta **can** be thin sectioned if it is done by someone with the proper experience and who is using the right kind of diamond knife. We have offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for some number of years; see URL http://www.2spi.com/catalog/standards/aem.html
The Ta system that prompted the original posting was thicker, however, it is also porous, and if properly infiltrated with resin, and assuming the porosity is above some point, in principle, at least, there is no reason why it could not be thin sectioned for TEM. No method is really artifact free, but some artifacts are more easy to recognize than others. Knife induced artifacts are anisotripic (e.g. directional) in nature and can be more easily recognized (as artifacts) than artifacts caused by say, ion milling, which are isotropic in nature.
When I say "right kind of diamond knife", I am not suggesting that the SPI diamond knife could do something above and beyond what other diamond knives could do. What I am saying is that there is a process of selection of the optimum knife angle, because one might have to be prepared to use a knife with a fairly low (for materials science work) knife angle, rather than a more blunt angle, but with the downside is that it will wear out more quickly. That might make for great business for a diamond knife supplier, but it does tend to get expensive for the user who is not so experienced with this kind of sectioning.
Disclaimer: SPI Supplies offers diamond knives, both materials and life science, and we have done this kind of sectioning, as a service, for commercial clients for over thirty years.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
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Following up on Jim's comments about using mechanical cutting and grinding techniques, I have some insight that may help. Jim mentioned that "There is no simple solution to the preparation of very hard and very soft materials in one specimen". This is true to some degree, but there are solutions. For example, the use of a wire saw is actually ideal for cutting through materials containing both hard and soft phases. The wire saw can be used either with an abrasive slurry or with a diamond impregnated wire. Use with an abrasive slurry is ideal for cutting materials of various hardness without damage or the smearing effect that you would see with a diamond wheel saw.
To be honest, I have no idea if this saw would do what you want it to do in this application, however if the difficulty is the hard/soft combination, the wire saw is one potentially viable solution. Also, I'm not sure if you need to embed the sample, but I would suggest forgoing tat step, if possible, if oyu are going to try the wire saw technique.
DISCLAIMER: South Bay Technology pioneered the development of the wire saw in the early 60s and continues to produce wire saws for many applications today.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } Tantalum is very hard and tough, so I doubt that it will section with either } diamond or tungsten carbide knives. Preparing the material like a geological } section by grinding is a possibility, but not a good one. Chances are that the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } Cheers } Jim Darley } Disclaimer: ProSciTech supplies both diamond and TC knives. I advised against } the use of either in this particular case. } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, June 06, 2001 1:54 AM, Maria Ericsson } [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core.
} } I have no idea how to section this stuff...! } } } } Thanks! } } Maria Ericsson } } } } My question is: } } I have a carbon coated porous tantalum matrix approximately 1 cm in } } circumference and 2 mm thick. This matrix has fixed cells adhered to the } } surface and throughout. I would like to embed this, make slides and stain } } for H&E to confirm the presences of cells as well as immunohistochemistry } } to characterize them. I am looking for suggestions on how I can section } } this matrix while maintaining its overall structure. Any information would } } be greatly appreciated. } } } } } } } } Lauri Wyner } } DF/HCC Central Pathology Cores Coordinator } } Harvard Medical School } } G1-126, Goldenson Building } } 220 Longwood Avenue } } Boston, MA 02115 } } Tele: (617) 432-4947 } } Fas: (617) 432-6474 } } Lauri_wyner-at-hms.harvard.edu } } } }
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
While infiltrating a set of specimens (insect abdomens and thoraxes) in LR White, something occurred that has so far defied my attempts to figure out. I was doing side by side microwave and conventional fixations, dehydrations, and embeddings. The microwave specimens were dehydrated in acetone, behaved normally, and we have polymerized blocks. However I dehydrated the conventional set in an ethanol series, as we have done many times before in LR White. The first infiltration step was 2 parts ethanol to one part LR White Medium Grade at room temp, and upon going later to change into 1:1, I found that the resin/ETOH mix had partially polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along with a few other choice words.
After the initial panic, I ended up putting the chunks into pure LR White from a newly opened bottle and the partially polymerized resin seemed to go back into solution. I ran them through a couple more changes of pure resin, then left them on a rocker overnight at room temperature. Checking them this morning, I found two of the samples were fine, and the third had polymerized into a rubbery mass. Same bottle of resin, same identically processed samples, same everything.
An additional tube with a sample of the "bad" resin was also happily unpolymerized, as was the remainder of the "bad" resin in the bottle, which I had left in the fume hood overnight at room temperature. The "bad" resin was slightly more than a year old and has been refrigerated at 4 C since we got it. The second resin is about 10 months old and has also been constantly refrigerated. Neither had any accelerator in them (at least none added by us).
I have my share of problems with LR White, but this one really has us puzzled. Could there have been something in the samples that triggered a polymerization? Can LR White react with some plastics in this way? (We were using a relatively new batch of Eppendorf tubes that seem more hydrophobic than our previous ones.) Could it have been the full moon?
Regards, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-8304 (573) 884-5414 (fax) email: tindallr-at-missouri.edu http://biotech.missouri.edu/emc
Does anyone have a reference for or some transmission electron micrographs that illustrate the effects of osmolarity of buffers on animal cells? I would like to show students on tour of our facility shrinkage and swelling of cells and organelles. I have other wonderful artifacts to show, including one of my first sections with chatter so bad it looks like mini-blinds...
Mahalo! Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Biomedical Electron Microscopy by Maunsbach A B & Afzelius B A (1999) has several pages of illustrations of shrinkage etc.
Dave
On Thu, 7 Jun 2001 16:29:12 -1000 (HST) Tina Carvalho {tina-at-pbrc.hawaii.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, All- } } Does anyone have a reference for or some transmission electron micrographs } that illustrate the effects of osmolarity of buffers on animal cells? I } would like to show students on tour of our facility shrinkage and swelling } of cells and organelles. I have other wonderful artifacts to show, } including one of my first sections with chatter so bad it looks like } mini-blinds... } } Mahalo! } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Chuck - the point of the original request was not to section Ta - which is hard enough, but do show the cells attached to the Ta. If you think that you can section Ta without the cells being ripped away, then do it. I will praise your skills when I can see useful results. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, June 08, 2001 8:26 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } } ==================================================== } Tantalum is very hard and tough, so I doubt that it will section with either } } diamond or tungsten carbide knives. Preparing the material like a geological } } section by grinding is a possibility, but not a good one. Chances are that } the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in } plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a } diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } ===================================================== } Actually Ta **can** be thin sectioned if it is done by someone with the } proper experience and who is using the right kind of diamond knife. We have } offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for } some number of years; see URL } http://www.2spi.com/catalog/standards/aem.html } } The Ta system that prompted the original posting was thicker, however, it is } also porous, and if properly infiltrated with resin, and assuming the } porosity is above some point, in principle, at least, there is no reason why } it could not be thin sectioned for TEM. No method is really artifact free, } but some artifacts are more easy to recognize than others. Knife induced } artifacts are anisotripic (e.g. directional) in nature and can be more } easily recognized (as artifacts) than artifacts caused by say, ion milling, } which are isotropic in nature. } } When I say "right kind of diamond knife", I am not suggesting that the SPI } diamond knife could do something above and beyond what other diamond knives } could do. What I am saying is that there is a process of selection of the } optimum knife angle, because one might have to be prepared to use a knife } with a fairly low (for materials science work) knife angle, rather than a } more blunt angle, but with the downside is that it will wear out more } quickly. That might make for great business for a diamond knife supplier, } but it does tend to get expensive for the user who is not so experienced } with this kind of sectioning. } } Disclaimer: SPI Supplies offers diamond knives, both materials and life } science, and we have done this kind of sectioning, as a service, for } commercial clients for over thirty years. } } Chuck } } PS: Please remember that we are nearly 100% paperless and we would ask that } any reply to this message be by way of the "reply" feature on your software, } so that the entire string of correspondence can come back to us and all be } in one place. }
} Hi, All- } } Does anyone have a reference for or some transmission electron micrographs } that illustrate the effects of osmolarity of buffers on animal cells? I } would like to show students on tour of our facility shrinkage and swelling } of cells and organelles. I have other wonderful artifacts to show, } including one of my first sections with chatter so bad it looks like } mini-blinds... } } Mahalo! } Tina
Arborgh et al. J. Ultrastruc. Res. 56:339-350, 1976. A classic study.
} } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The utility of rubber stoppers to seal a SEM has been doubted. 1 Somebody may chose too smaller stopper size and the stopper may be sucked into the column. Good point, just like noting that the stopper must be inserted from the outside of the SEM. Yes!
2 The X-ray objection is one which needs to be addressed. I had looked up that data some years ago, when in need of temporary blanks. Because of the questions I again looked for the data, now on the marvelous Internet . The common Monte Carlo programs would give similar results, but the show the bulk of the X-ray interaction, whereas we are interested in maximum penetration. I recommend http://www-cxro.lbl.gov/optical_constants/atten2.html This site allows some data entry and then calculates the attenuation length at which the X-ray intensity falls of to 1eV at the surface. Assuming that Teflon is close to rubber in absorption, then 20keV X-ray photons are attenuated in 5mm of rubber. Whereas 30keV photons are only attenuated in about 12mm of rubber. Clearly, it would be unwise to run the kV of a TEM, when blanked with a rubber stopper. It is also clear that a SEM/ Probe when operated at 20 kV would not produce X-rays capable of penetrating a normal rubber stopper. Incidentally, a 20kV beam would produce somewhat lower KeV X-ray photons. X-ray generation depends on the interaction with electron shells. Only the heavier metals can produce powerful X-rays and to reach full fluorescence (maximum production) the kV needs to be about 2x that of the X-ray KeV. Cheers Jim Darley PS Red stoppers contain Iron oxide pigment, whereas black stoppers (I presume) have negligible metal content. Question: Considering that iron has greater X-ray stopping power than carbon, should we assume that it is smarter to use red stoppers??? The opposing argument would be that the iron could generate more high energy X-rays.
I don't know the answer (would be amused to learn though), but I prefer silicone rubber stoppers because they form a better vacuum seal.
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, June 07, 2001 10:14 PM, Marie E. Cantino [SMTP:cantino-at-uconnvm.uconn.edu] wrote: } } } Just out of curiousity. . . how much X-radiation penetrates the rubber } stopper? } } Marie }
} I vote with Ritchie. Rubber stoppers are great for temporary problems, } particularly for finding vacuum leaks. One stopper can eliminate a more complex } assembly. I somehow prefer the silicone rubber stoppers. } However, do consider X-ray penetration. Particularly on instruments above } 20kV. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com
} } } } } A rubber stopper, well-greased with appropriate grease, worked fine } } on my 840 in a similar situation, no degradation of vacuum. } } Just make absolutely sure that it's big enough that there's no chance } } of its being sucked right in, and that nobody knocks into it! } } } } cheers } } } } rtch } } } } Dr. Marie E. Cantino } Dept. of Physiology and Neurobiology, U-2131 } University of Connecticut } Storrs, CT 06269-2131 } Phone: 860-486-3588 } Fax: 860-486-6369 }
I usually wrap several layers of aluminum foil over the stopper if I need to use it. This will stop most of the X-rays. Fortunately, the EDS port is to the rear of the column and pointing upward.
Earl
----- Original Message ----- } From: "Arthur Day" {ard-at-ansto.gov.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, June 07, 2001 9:53 PM
-------- Original Message --------
Pehaps someone could provide a better analysis, but I have a couple of anecdotes:
I had a Canadian customer (20+ years ago) who specialized in very long EDS x-ray sweeps in either dot map form or a combined YZ modulated form. As I recall, some of these scans were up to 16 hours long. He had a Plexiglass port cover so that he could easily see his chamber geometry. This had a metal cover that was velvet lined for light leakage. Once he put his radiation badge inside the metal cover for a month, then turned it in. There apparently was nothing out of the ordinary, as his rad peo;le never said "boo". Perhaps it's a commentary on the rad people, but a full month exposure?
Second, I had another customer get me some rad figures on an SEM gun operating at 30kV and 150uA (1.5x10E-4). The bottom line was that the x-ray output from the gun was very serious, but all the guns are enclosed in steel. No problem. At the specimen for EDS work we're usually talking about 200-600pA (2-6x10E-10), or roughly 6 orders of magnitude less current and the stopper is going to absorb some of the x-rays generated. Is this really a problem?
I'm not talking about TEMs, as they can be very dangerous. The current at the screen is much higher, the excitation voltage is much higher and there is this large expanse of glass that MUST be leaded. Perhaps the currents used for WDS would also present a problem (10-100 nA or 10E-8 to 10E-7) at 2 to 3 orders of magnitude more than EDS.
Don't forget, those nice color displays on the four computers surrounding you in your lab also generate x-rays and their beam currents are in the mA range (10E-3) and an accelerating voltage of about 25kV. There may be a decelerating grid and leaded glass on the front, but I think you will find the the shielding on the rear of the monitor is not so rigorous, Your dentist's x-rays are about 70kV and only one order of magnitude greater current.
I'd love some feed-back from someone who knows radiation because I've felt that applying TEM rules to SEMs is gross over-kill, especially with so many color monitors around. How often do you have your decelerating grid checked for proper operation? If it doesn't operate correctly, your exposure could be very dangerous, given the time that one sits in front of these things and their distance from your face.
If I'm way off base, I'd like to know and know why.
Ken Converse owner Quality Images third party SEM service Delta, PA
Arthur Day wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Just out of curiousity. . . how much X-radiation penetrates the rubber } } stopper? } } } } Marie } } } } Doesn't anybody supply X-ray proof rubber bungs for microscope ports? } } No worries. Just wrap a few sheets of lead foil around the column and } make sure pregnant operators stay below 5kV ;-( } } Surely something somewhat less desperate could be cobbled together out } of metal without too much extra work? } } } }
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Jim Darley wrote:
==================================================== Tantalum is very hard and tough, so I doubt that it will section with either
diamond or tungsten carbide knives. Preparing the material like a geological
section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and
view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. ===================================================== Actually Ta **can** be thin sectioned if it is done by someone with the proper experience and who is using the right kind of diamond knife. We have offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for some number of years; see URL http://www.2spi.com/catalog/standards/aem.html
The Ta system that prompted the original posting was thicker, however, it is also porous, and if properly infiltrated with resin, and assuming the porosity is above some point, in principle, at least, there is no reason why it could not be thin sectioned for TEM. No method is really artifact free, but some artifacts are more easy to recognize than others. Knife induced artifacts are anisotripic (e.g. directional) in nature and can be more easily recognized (as artifacts) than artifacts caused by say, ion milling, which are isotropic in nature.
When I say "right kind of diamond knife", I am not suggesting that the SPI diamond knife could do something above and beyond what other diamond knives could do. What I am saying is that there is a process of selection of the optimum knife angle, because one might have to be prepared to use a knife with a fairly low (for materials science work) knife angle, rather than a more blunt angle, but with the downside is that it will wear out more quickly. That might make for great business for a diamond knife supplier, but it does tend to get expensive for the user who is not so experienced with this kind of sectioning.
Disclaimer: SPI Supplies offers diamond knives, both materials and life science, and we have done this kind of sectioning, as a service, for commercial clients for over thirty years.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
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Randy: This has happened a bunch of times to me. It only happens with osmicated tissues. I have run up the same tisse +/- osmication and premature polymerization only occurs in the osmicated ones - not every time or at the same rate. I guess there is something that doesn't get rinsed out that can trigger it - perhaps more so when the LRW is aging. I am starting a LRW infiltration with osmicated tissue using a brand new bottle of LRW and I will let you know what happens. my tissues were in glass vials so it isn't the plastic tubes. it has happened at both 4 C and room temp. i have a vague recollection posting this on the microscopy listserver and not getting much of a response. maddening problem, isn't it?
} ------------------------------------------------------------. } } } Dear Listers, } } While infiltrating a set of specimens (insect abdomens and thoraxes) in LR } White, something occurred that has so far defied my attempts to figure out. } I was doing } side by side microwave and conventional fixations, dehydrations, and } embeddings. The microwave specimens were dehydrated in acetone, behaved } normally, and we } have polymerized blocks. However I dehydrated the conventional set in an } ethanol series, as we have done many times before in LR White. The first } infiltration step } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } upon going later to change into 1:1, I found that the resin/ETOH mix had } partially } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along } with a few other choice words. } } After the initial panic, I ended up putting the chunks into pure LR White } from a newly opened bottle and the partially polymerized resin seemed to go } back into solution. I } ran them through a couple more changes of pure resin, then left them on a } rocker overnight at room temperature. Checking them this morning, I found } two of the } samples were fine, and the third had polymerized into a rubbery mass. } Same bottle of resin, same identically processed samples, same everything. } } An additional tube with a sample of the "bad" resin was also happily } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } I had left in the fume } hood overnight at room temperature. The "bad" resin was slightly more } than a year old and has been refrigerated at 4 C since we got it. The } second resin is about 10 } months old and has also been constantly refrigerated. Neither had any } accelerator in them (at least none added by us). } } I have my share of problems with LR White, but this one really has us } puzzled. Could there have been something in the samples that triggered a } polymerization? Can } LR White react with some plastics in this way? (We were using a } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } previous ones.) Could } it have been the full moon? } } Regards, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } (573) 882-8304 } (573) 884-5414 (fax) } email: tindallr-at-missouri.edu } http://biotech.missouri.edu/emc }
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
You might have nailed it. These were osmicated tissues, since they weren't intended for immuno. Our client prefers LR White for making thick sections for light microscopy, so these specimens were prepared with osmium and UA post-fixations. Given the large size and difficult nature of insect parts, it's very possible that something remained in the tissue despite several lengthy washes. At least we have back-up specimens!
Thanks for the response. At least it's something to work with.
Randy
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, June 08, 2001 9:24 AM To: Tindall, Randy D. Cc: Microscopy-at-msa.microscopy.com
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Colleagues....
I hate to say this, but alot of mail got rejected today by accident.
I was updating the junk mail filters last night and unintentionally created a new filter which rejected the majority of the mail posted today.
If you received a rejection message indicating that you had a subject line of \b$ please repost your message. It was not your fault, but mine.
Listers, Thanks to all who responded to my particle size analysis posting recently. Reasons, including a heavy workload, would not allow us to accept the project at this time. We now have several resources from which to choose. I appreciate the time and effort from those who offered constructive answers. From the one who offered questionable musings... contact me off-line and I'll be more
specific about what can be done with those. Winston ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor June 8, 2001 10:07 AM Cannon Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 (Ship to: 1000 Blythe Blvd ) Fax: 704-355-0589 Charlotte, NC 28232-2861 (Ship to: 28203 ) WWiggins-at-Carolinas.org {mailto:WWiggins-at-Carolinas.org} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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I also have a number of old Leitz "Technical Information Bulletins" that will be listed later.
Hope there is a use.
FCM
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
I concluded from the original information given that the tantalum matrix the young lady was speaking about was Cytomatrix (http://www.cytomatrix.com). The matrix is reportedly a tantalum coated graphite core material that was first use as a support for bone marrow/bone cell regeneration. That's why I suggested the glycol methacrylate, though you may still be correct about it's 'hardness'.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Jim at Proscitech } Reply To: jim-at-proscitech.com } Sent: Thursday, June 7, 2001 6:25 AM } To: 'Maria Ericsson'; Microscopy-at-sparc5.microscopy.com } Subject: RE: sectioning tantalum matrix } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Tantalum is very hard and tough, so I doubt that it will section with } either } diamond or tungsten carbide knives. Preparing the material like a } geological } section by grinding is a possibility, but not a good one. Chances are that } the } grinding material would fill the voids and the tissue would be ground away } } first (although ProSciTech and others supply diamond grit embedded in } plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a } diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic } and } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } Cheers } Jim Darley } Disclaimer: ProSciTech supplies both diamond and TC knives. I advised } against } the use of either in this particular case. } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, June 06, 2001 1:54 AM, Maria Ericsson } [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core. } } I have no idea how to section this stuff...! } } } } Thanks! } } Maria Ericsson } } } } My question is: } } I have a carbon coated porous tantalum matrix approximately 1 cm in } } circumference and 2 mm thick. This matrix has fixed cells adhered to the } } surface and throughout. I would like to embed this, make slides and } stain } } for H&E to confirm the presences of cells as well as } immunohistochemistry } } to characterize them. I am looking for suggestions on how I can section } } this matrix while maintaining its overall structure. Any information } would } } be greatly appreciated. } } } } } } } } Lauri Wyner } } DF/HCC Central Pathology Cores Coordinator } } Harvard Medical School } } G1-126, Goldenson Building } } 220 Longwood Avenue } } Boston, MA 02115 } } Tele: (617) 432-4947 } } Fas: (617) 432-6474 } } Lauri_wyner-at-hms.harvard.edu } } } } } } }
I agree with David, there are several ways to cut and grind samples with multiple materials of varying hardnesses. The EXAKT Cutting/Grinding Systems were developed just for this purpose. One of the major issues in cutting or grinding these types of materials is the force required to cut the hard material vs. the force required to cut the softer material. Like the diamond wire saw a diamond embedded band system designed to section with a minimum of force can also achieve excellent results. The reduced force technology (reduction of the active cutting or grinding surface) is not new but has new applications in material research. And the technology can be applied to the grinding process as well to achieve flat surfaces at material interfaces.
Many of these techniques have been developed for LM in the biomaterials industry where the materials can vary from cobalt chrome implanted in bone or bone cement to memory metal stents implanted in soft arteries. The technology works equally well for rubber bonded to metal or solders layered on ceramics or carbon fibers embedded in unpolymerized resin. One of the key features of reducing the force and changing the cutting or grinding direction is also to prevent smearing of one material over an interface. Biomaterials researchers have been the driving force in developing this technology for the past 15 years and for LM it is a viable alternative to standard metallographic preparations.
Disclaimer: EXAKT Technologies, Inc. is the North American Distributor for EXAKT Apparatebau the manufacturer of various cutting devices and grinders for multiple material applications.
Linda Durbin
-----Original Message----- } From: David Henriks [mailto:henriks-at-southbaytech.com] Sent: Thursday, June 07, 2001 7:48 PM To: Microscopy Listerver
Following up on Jim's comments about using mechanical cutting and grinding techniques, I have some insight that may help. Jim mentioned that "There is no simple solution to the preparation of very hard and very soft materials in one specimen". This is true to some degree, but there are solutions. For example, the use of a wire saw is actually ideal for cutting through materials containing both hard and soft phases. The wire saw can be used either with an abrasive slurry or with a diamond impregnated wire. Use with an abrasive slurry is ideal for cutting materials of various hardness without damage or the smearing effect that you would see with a diamond wheel saw.
To be honest, I have no idea if this saw would do what you want it to do in this application, however if the difficulty is the hard/soft combination, the wire saw is one potentially viable solution. Also, I'm not sure if you need to embed the sample, but I would suggest forgoing tat step, if possible, if oyu are going to try the wire saw technique.
DISCLAIMER: South Bay Technology pioneered the development of the wire saw in the early 60s and continues to produce wire saws for many applications today.
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} } Tantalum is very hard and tough, so I doubt that it will section with either } diamond or tungsten carbide knives. Preparing the material like a geological } section by grinding is a possibility, but not a good one. Chances are that the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } Cheers } Jim Darley } Disclaimer: ProSciTech supplies both diamond and TC knives. I advised against } the use of either in this particular case. } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, June 06, 2001 1:54 AM, Maria Ericsson } [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core.
} } I have no idea how to section this stuff...! } } } } Thanks! } } Maria Ericsson } } } } My question is: } } I have a carbon coated porous tantalum matrix approximately 1 cm in } } circumference and 2 mm thick. This matrix has fixed cells adhered to the } } surface and throughout. I would like to embed this, make slides and stain } } for H&E to confirm the presences of cells as well as immunohistochemistry } } to characterize them. I am looking for suggestions on how I can section } } this matrix while maintaining its overall structure. Any information would } } be greatly appreciated. } } } } } } } } Lauri Wyner } } DF/HCC Central Pathology Cores Coordinator } } Harvard Medical School } } G1-126, Goldenson Building } } 220 Longwood Avenue } } Boston, MA 02115 } } Tele: (617) 432-4947 } } Fas: (617) 432-6474 } } Lauri_wyner-at-hms.harvard.edu } } } }
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
I have have a task of quantifying the actin distribution in cultured endothelial cells and subsequently comparing different culture conditions. So far I have managed to isolate the filaments and threshold them. Now...I wonder what is the best way to put a number on the distribution within each cell. One possibility I was tinking of, is using a sobel operator to give the direction of the lines representing the filaments and then display the data as histograms of filament orientations.
Has anyone done this or know of a good wayto go about this type of analysis?
We have been using an older Balzer's sputter coater to coat SEM samples with Pt. Lately the grain structure of the Pt coating has become much more obvious (at around 100,000x.) Any experts have a thought on why this is occurring and how to improve the process? Thanks for any guidelines and help. Regards, Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
I did some looking into costs/hr and finally came to the conclusion that if I made an assumption that well funded institutions with shared facilities had set prices for instrument use, it probably reflected what the market - i.e. the granting agencies - would bear. What I discovered was prices that varied but stayed in the range of $25 to $35. $29 seemed to be popular, though I wondered why not the infinitely more popular prince of $29.95.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com } Sent: Thursday, June 7, 2001 8:03 AM } To: microscopy-at-sparc5.microscopy.com } Subject: EM COSTING } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } we are in the process of cost our EM service, that is a cost breakdown } for } doing EM. i know the actual costs are very small. i was wondering if } anyone } has done it recently. we are a diagnostic lab. } john hoffpauir } cooper hospital } }
A little more insight on Xray absorption of materials: Twenty years ago when I was doing EDS XRF, I did a lot of work on Xray filters on the primary Xray beam from Xray tubes. I designed both edge filters (the absorption edges of each element can be used to strongly remove X-rays above certain energies} and "white" filters to remove all Xrays below certain energies. My white filters were usually layers of filter paper (no Ti pigment). My experience was that at below 30KV on the Xray tube almost nothing would get through 1/2 inch of organic materials. Above 30 KeV we had worry about X-ray leakage. The Xrays tubes (and Electron microscopes) give off both primary lines and bremsstrahlung xrays. The Bremsstrahlung peak intensity energy in KeV by rule of thumb is about 2/3 the energy of the electron beam. You may observe in your SEM by EDS the X-ray energy spectrum. It is that background spectrum. Since I had a spectrometer I could ready see what energies passed or started leaking through my filters. You too can do the Xray absorption spectrum test inside your chamber by placing the stopper or other material in front of your EDS detector window and see what comes through. The EDS detector is much more sensitive than a Geiger counter or film.
Therefore, if the SEM is used below 30 kV, then a thick rubber stopper will absorb the X-rays. Still worried? Stick a radiation badge on the outside of the stopper to check.
Ronald Vane XEI Scientific
-----Original Message----- } From: Ken Converse {qualityimages-at-netrax.net} To: Arthur Day {ard-at-ansto.gov.au} ; MSA, listserver {Microscopy-at-sparc5.microscopy.com}
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Tina Carvalho wrote:
} Hi, All- } } Does anyone have a reference for or some transmission electron micrographs } that illustrate the effects of osmolarity of buffers on animal cells? I } would like to show students on tour of our facility shrinkage and swelling } of cells and organelles. I have other wonderful artifacts to show, } including one of my first sections with chatter so bad it looks like } mini-blinds... } } Mahalo! } Tina
Arborgh et al. J. Ultrastruc. Res. 56:339-350, 1976. A classic study.
} } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
You can try JB-4 embedding media instead of LR White. It works pretty well for thick sectioning and maitains excellent structure preservation.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Fri, 8 Jun 2001, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Tom, } } You might have nailed it. These were osmicated tissues, since they weren't } intended for immuno. Our client prefers LR White for making thick sections } for light microscopy, so these specimens were prepared with osmium and UA } post-fixations. Given the large size and difficult nature of insect parts, } it's very possible that something remained in the tissue despite several } lengthy washes. At least we have back-up specimens! } } Thanks for the response. At least it's something to work with. } } Randy } } -----Original Message----- } } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] } Sent: Friday, June 08, 2001 9:24 AM } To: Tindall, Randy D. } Cc: Microscopy-at-msa.microscopy.com } Subject: Re: TEM: LR White weirdness } } } Randy: This has happened a bunch of times to me. It only happens } with osmicated tissues. I have run up the same tisse +/- osmication } and premature polymerization only occurs in the osmicated ones - not } every time or at the same rate. I guess there is something that } doesn't get rinsed out that can trigger it - perhaps more so when the } LRW is aging. I am starting a LRW infiltration with osmicated tissue } using a brand new bottle of LRW and I will let you know what happens. } my tissues were in glass vials so it isn't the plastic tubes. it has } happened at both 4 C and room temp. i have a vague recollection } posting this on the microscopy listserver and not getting much of a } response. maddening problem, isn't it? } } } ------------------------------------------------------------. } } } } } } Dear Listers, } } } } While infiltrating a set of specimens (insect abdomens and thoraxes) in } LR } } White, something occurred that has so far defied my attempts to figure out. } } I was doing } } side by side microwave and conventional fixations, dehydrations, and } } embeddings. The microwave specimens were dehydrated in acetone, behaved } } normally, and we } } have polymerized blocks. However I dehydrated the conventional set in } an } } ethanol series, as we have done many times before in LR White. The first } } infiltration step } } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } } upon going later to change into 1:1, I found that the resin/ETOH mix had } } partially } } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, } along } } with a few other choice words. } } } } After the initial panic, I ended up putting the chunks into pure LR } White } } from a newly opened bottle and the partially polymerized resin seemed to go } } back into solution. I } } ran them through a couple more changes of pure resin, then left them on } a } } rocker overnight at room temperature. Checking them this morning, I found } } two of the } } samples were fine, and the third had polymerized into a rubbery mass. } } Same bottle of resin, same identically processed samples, same everything. } } } } An additional tube with a sample of the "bad" resin was also happily } } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } } I had left in the fume } } hood overnight at room temperature. The "bad" resin was slightly more } } than a year old and has been refrigerated at 4 C since we got it. The } } second resin is about 10 } } months old and has also been constantly refrigerated. Neither had any } } accelerator in them (at least none added by us). } } } } I have my share of problems with LR White, but this one really has us } } puzzled. Could there have been something in the samples that triggered a } } polymerization? Can } } LR White react with some plastics in this way? (We were using a } } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } } previous ones.) Could } } it have been the full moon? } } } } Regards, } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } (573) 882-8304 } } (573) 884-5414 (fax) } } email: tindallr-at-missouri.edu } } http://biotech.missouri.edu/emc } } } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } }
At 12:55 PM 6/8/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I am considering acquiring a cryostat for the laboratory. Does anyone have an experience, positive or negative, with the Bright OTF cryostat? Please respond to me offline.
I am in need of a reference EELS spectum of ZnO (or probably any other Zn[+2] compound ) to help me interpret some data. I've already checked the CEMES EELS database to no avail, and I've done some web-searching with lots of hits but mostly to conference abstracts but no hard data. I'm attempting to distinguish between Zn metal, for which I have a reference EELS spectrum, and Zn[+2}.
Tia, Mike Nesson-- -- _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu
You are right in the comparison of SEMs and TEMs. The accelerating voltages typically used in TEMs creates a much greater concern as do the glass viewing ports directly open to the electron beam - sample interactions. In fact, the majority of problems in TEMs have been inadequate lead content of the glass in those ports. SEMs are inherently safe from external emissions in that their design results in a generous metallic shielding surrounding all portions that can generate x-rays.
However, even in SEMs, there can be exceptions. Remember the small aluminum plug in the columns of ETEC SEMs, the one opposite where the scan coil cables enter? If that plug is not in place, then there is a potential for external radiation. While the designs may be inherently safe, there is still room for error.
Many states require a regular scan for external radiation on any x-ray generating equipment. While I have heard of quite a few measurable responses on TEMs (including one just two days ago at a DOE site), I have never heard of any detectable radiation from an SEM.
Regarding computer monitors, don't worry. While early TV sets did produce considerable x-rays externally, modern TVs and computer monitors really don't. Unless you plan on spending 24 hours a day hugging the back side of a monitor, I wouldn't worry.
Finally, regarding the temporary use of a rubber stopper - It shouldn't pose a problem. Most of the ports on an SEM, and particularly those for EDS systems, are on the rear of the chamber. SEMs, given their usual operating conditions and limitations, are not prodigious x-ray producers. Most would be absorbed by the thickness of a stopper capable of withstanding the vacuum pressures, and what little could escape would be directed away from anyone using the instrument. Since any resulting radiation would be attenuated by the square of the distance from the source (in this case the x-rays penetrating the stopper) there would normally be no detectable exposure except, perhaps, at the surface of the stopper.
I do have a problem with permanent plastic viewports, however. These are generally placed on the sides of the sample chamber where a more direct path to the operator is possible. Whenever I find a customer with a plastic viewport, I always strongly suggest that they keep a suitable cover over it when they aren't being used to view the sample presentation. Surprisingly thin ports can be made of materials like polycarbonate that are capable of withstanding atmospheric pressures.
The real questions should be, if this is a situation that may occur from time to time, should a proper port cover be fabricated and kept on hand so that there are no such questions, now or in the future?
On Friday, June 08, 2001 9:16 AM, Ken Converse [SMTP:qualityimages-at-netrax.net] wrote: } } Pehaps someone could provide a better analysis, but I have a couple of } anecdotes: } } I had a Canadian customer (20+ years ago) who specialized in very long } EDS x-ray sweeps in either dot map form or a combined YZ modulated } form. As I recall, some of these scans were up to 16 hours long. He } had a Plexiglass port cover so that he could easily see his chamber } geometry. This had a metal cover that was velvet lined for light } leakage. Once he put his radiation badge inside the metal cover for a } month, then turned it in. There apparently was nothing out of the } ordinary, as his rad peo;le never said "boo". Perhaps it's a commentary } on the rad people, but a full month exposure? } } Second, I had another customer get me some rad figures on an SEM gun } operating at 30kV and 150uA (1.5x10E-4). The bottom line was that the } x-ray output from the gun was very serious, but all the guns are } enclosed in steel. No problem. At the specimen for EDS work we're } usually talking about 200-600pA (2-6x10E-10), or roughly 6 orders of } magnitude less current and the stopper is going to absorb some of the } x-rays generated. Is this really a problem? } } I'm not talking about TEMs, as they can be very dangerous. The current } at the screen is much higher, the excitation voltage is much higher and } there is this large expanse of glass that MUST be leaded. Perhaps the } currents used for WDS would also present a problem (10-100 nA or 10E-8 } to 10E-7) at 2 to 3 orders of magnitude more than EDS. } } Don't forget, those nice color displays on the four computers } surrounding you in your lab also generate x-rays and their beam currents } are in the mA range (10E-3) and an accelerating voltage of about 25kV. } There may be a decelerating grid and leaded glass on the front, but I } think you will find the the shielding on the rear of the monitor is not } so rigorous, Your dentist's x-rays are about 70kV and only one order of } magnitude greater current. } } I'd love some feed-back from someone who knows radiation because I've } felt that applying TEM rules to SEMs is gross over-kill, especially with } so many color monitors around. How often do you have your decelerating } grid checked for proper operation? If it doesn't operate correctly, } your exposure could be very dangerous, given the time that one sits in } front of these things and their distance from your face. } } If I'm way off base, I'd like to know and know why. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Arthur Day wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Just out of curiousity. . . how much X-radiation penetrates the rubber } } } stopper? } } } } } } Marie } } } } } } } Doesn't anybody supply X-ray proof rubber bungs for microscope ports? } } } } No worries. Just wrap a few sheets of lead foil around the column and } } make sure pregnant operators stay below 5kV ;-( } } } } Surely something somewhat less desperate could be cobbled together out } } of metal without too much extra work? } } } } } } } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} I have in my possession the following [Old Microscope Catalogs]: } I also have a number of old Leitz "Technical Information Bulletins" that } will be listed later. } Hope there is a use. } Fred -
If you don't get takers for all of these, there's a microscopy book dealer in England who can probably find them a home: http://www.savonabooks.free-online.co.uk It would be a pity to just trash them...
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I know someone makeing a CDROM of Leitz information that is not copyrighted. It is going to be a very reasonably priced commerical project.
I would post his name but I am in the middle of changeing computers and I don't have his email address at hand.
Gordon Couger
----- Original Message ----- } From: "Caroline Schooley" {schooley-at-mcn.org} To: "Monson, Frederick C." {fmonson-at-wcupa.edu} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, June 09, 2001 10:32 AM
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Engines can at any time delay their indexing for maintenance and many other reasons.
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Unwanted polymerisation of LR White resin during infiltration is an occasional problem with the resin. I have never experienced it with fresh batches, indeed the only times I have had this problem was with batches over 1 year old (the recommended use by time). An possible explanation was suggested to me by Roy Gillett of London Resin a number of years ago:
Quote "The reason this pre-polymerisation occurs only with tissue must be something to do with a tissue constituent catalysing polymerisation. Older resin is much more susceptibe to this that fresh monomer becaue of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenousd catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
We have used LR White resin extensively for a long time - perhaps the reason we don't have a problem is that our resin never gets anything like a year old before we use it/
Best Wishes
Ian
} } Dear Listers, } } While infiltrating a set of specimens (insect abdomens and thoraxes) in LR } White, something occurred that has so far defied my attempts to figure out. } I was doing } side by side microwave and conventional fixations, dehydrations, and } embeddings. The microwave specimens were dehydrated in acetone, behaved } normally, and we } have polymerized blocks. However I dehydrated the conventional set in an } ethanol series, as we have done many times before in LR White. The first } infiltration step } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } upon going later to change into 1:1, I found that the resin/ETOH mix had } partially } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along } with a few other choice words. } } After the initial panic, I ended up putting the chunks into pure LR White } from a newly opened bottle and the partially polymerized resin seemed to go } back into solution. I } ran them through a couple more changes of pure resin, then left them on a } rocker overnight at room temperature. Checking them this morning, I found } two of the } samples were fine, and the third had polymerized into a rubbery mass. } Same bottle of resin, same identically processed samples, same everything. } } An additional tube with a sample of the "bad" resin was also happily } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } I had left in the fume } hood overnight at room temperature. The "bad" resin was slightly more } than a year old and has been refrigerated at 4 C since we got it. The } second resin is about 10 } months old and has also been constantly refrigerated. Neither had any } accelerator in them (at least none added by us). } } I have my share of problems with LR White, but this one really has us } puzzled. Could there have been something in the samples that triggered a } polymerization? Can } LR White react with some plastics in this way? (We were using a } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } previous ones.) Could } it have been the full moon? } } Regards, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } (573) 882-8304 } (573) 884-5414 (fax) } email: tindallr-at-missouri.edu } http://biotech.missouri.edu/emc } }
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
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SEARCH ENGINES DELIVER TARGETED UNIQUE TRAFFIC TO WEB SITE
If you don't find what you're looking for in the first 10 to 40 matches ON A SEARCH ENGINE, what do you do? If you are like most people, you simply go to the next search engine and try again.
If your site is not coming up in the TOP 40 matches On a search engine, you're losing MONEY!
IT 'S THAT SIMPLE!
Improving search engine TRAFFIC means:
MORE HITS MORE BUSINESS MORE SUCCESS
The most important advertising dollar being spent should be for search engine TRAFFIC.
1. Approximately 95% of all Internet users start with a search engine query.
2. Anyone who comes to your site from a major search engine is 100 times more likely to become a customer because they were specifically looking for your product, goods or services.
3. Search engine traffic gives you substantially more for your advertising dollar than banners. Moreover Banners are done on impressions. An impression is just someone that went to a web page with your Banner ad on the page. It does not mean your Ad was seen.
You Can't Get Your Companies Web Site Indexed by the Search Engines?
Unfortunately, this is all too common of a Problem. You're not the only one frustrated about the length of time it takes to be listed, or all the pitfalls involved. It takes anywhere from 2 days to as much as 6 months to be listed on all the search engines. Waiting several weeks to months can be extremely frustrating.
WHEN TO DO YOUR SUBMISSION?
Engines can at any time delay their indexing for maintenance and many other reasons.
So do you feel lucky? Do YOU?
Lucky if you submit A PAGE just days before an engine does a complete refresh of their many indexes/databases.
WE Know exactly how each search engine works, and we know when to submit and what to submit. Search engines are changing Daily and we study them each day. Your competitors ARE -at- the mercy of OUR Marketing Departments. 6 Plus Years Now in the search engine wars, We are Masters of words like: MP3 - BOOKS - WEB SITE HOSTING - MARKETING - - FREE WEB SITES - CASINO - - CASINO REVIEWS - BALLS - - LOGOS - ART - ATTORNEY'S - NEW CAR PARTS - OLD CAR PART - - NETWORK MARKETING - WATER FILTERS - - SCALES - AND THE LIST GOES ON - -
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THE CYBER TRAFFIC TECHNICIANS.
You need is us in your corner, we will take control of the submission cycle of your domain/s. We Provide you with: a real report that shows you what's been done and when completed, we offer free rank reporting.
The cost is 79.00 US Dollars month to month - 69.00 US Dollars a month when paying for 3 months at a time - 59.00 US Dollars a month when paying for 12 months. This Price is good for the next 5 orders only. DON'T delay in ordering.
If You fax a check, there is no need for you to mail the original. We will draft a new check, with the exact information from your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "CYBERCONTROL"
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I have a question about measuring small cellular structures at the light microscope level using Argus 10 enhancement and measurement software.
I am measuring vesicle diameters in living cells of the fungus Neurospora. They measure about 350-400 nm in diameter using the Argus 10 software. With cryofixed TEMs of the same cell type, the only vesicles with the same distribution as at the LM level measure 150 nm diameter and there are no structures at 350-400 nm.
All my calibrations of both the LM and TEM have been checked and double-checked.
It is my understanding that video-enhanced LM can amplify signal (or sizes) so that structures below limit of LM resolution can be detected (eg, imaging of individual microtubules with DIC/VELM). I am wondering if this is the source of this discrepancy that it see and if there are references that describe this.
Thanks so much,,,,,, Robby Roberson
*************************************** Robert W. Roberson, PhD Department of Plant Biology Molecular and Cellular Biology Program Arizona State University Tempe, AZ 85282 Office/Lab 480-965-8618
At 7:53 AM -0700 6/8/01, Eric wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
We have notice a strange effect of colored Microcentrifuge tubes. The colored tubes will cause hardening of the LR White at room temperature. Something in the coloring dyein the tubes must be acting as a catalyst.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: IAN HALLETT [mailto:ihallett-at-hortresearch.co.nz] Sent: Sunday, June 10, 2001 3:35 PM To: microscopy-at-sparc5.microscopy.com
Randy and others:
Unwanted polymerisation of LR White resin during infiltration is an occasional problem with the resin. I have never experienced it with fresh batches, indeed the only times I have had this problem was with batches over 1 year old (the recommended use by time). An possible explanation was suggested to me by Roy Gillett of London Resin a number of years ago:
Quote "The reason this pre-polymerisation occurs only with tissue must be something to do with a tissue constituent catalysing polymerisation. Older resin is much more susceptibe to this that fresh monomer becaue of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenousd catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
We have used LR White resin extensively for a long time - perhaps the reason we don't have a problem is that our resin never gets anything like a year old before we use it/
Best Wishes
Ian
} } Dear Listers, } } While infiltrating a set of specimens (insect abdomens and thoraxes) in LR } White, something occurred that has so far defied my attempts to figure out. } I was doing } side by side microwave and conventional fixations, dehydrations, and } embeddings. The microwave specimens were dehydrated in acetone, behaved } normally, and we } have polymerized blocks. However I dehydrated the conventional set in an } ethanol series, as we have done many times before in LR White. The first } infiltration step } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } upon going later to change into 1:1, I found that the resin/ETOH mix had } partially } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along } with a few other choice words. } } After the initial panic, I ended up putting the chunks into pure LR White } from a newly opened bottle and the partially polymerized resin seemed to go } back into solution. I } ran them through a couple more changes of pure resin, then left them on a } rocker overnight at room temperature. Checking them this morning, I found } two of the } samples were fine, and the third had polymerized into a rubbery mass. } Same bottle of resin, same identically processed samples, same everything. } } An additional tube with a sample of the "bad" resin was also happily } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } I had left in the fume } hood overnight at room temperature. The "bad" resin was slightly more } than a year old and has been refrigerated at 4 C since we got it. The } second resin is about 10 } months old and has also been constantly refrigerated. Neither had any } accelerator in them (at least none added by us). } } I have my share of problems with LR White, but this one really has us } puzzled. Could there have been something in the samples that triggered a } polymerization? Can } LR White react with some plastics in this way? (We were using a } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } previous ones.) Could } it have been the full moon? } } Regards, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } (573) 882-8304 } (573) 884-5414 (fax) } email: tindallr-at-missouri.edu } http://biotech.missouri.edu/emc } }
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
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I would like to be of some assistance on the subject of a solution to:-'There is no simple solution to the preparation of very hard and very soft materials in one specimen.'
As our Bright/Hacker OTF/AS/D/LT cryostat is able to section a hard and soft specimen incorporated into one specimen. If this is of interest I would be pleased to receive sample and return our results to those who have a problem.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England
Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: AlanBright-at-brightinstruments.com Web Site: www.brightinstruments.com
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: 07 June 2001 23:26 To: MICROSCOPY BB
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Jim Darley wrote:
==================================================== Tantalum is very hard and tough, so I doubt that it will section with either
diamond or tungsten carbide knives. Preparing the material like a geological
section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and
view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. ===================================================== Actually Ta **can** be thin sectioned if it is done by someone with the proper experience and who is using the right kind of diamond knife. We have offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for some number of years; see URL http://www.2spi.com/catalog/standards/aem.html
The Ta system that prompted the original posting was thicker, however, it is also porous, and if properly infiltrated with resin, and assuming the porosity is above some point, in principle, at least, there is no reason why it could not be thin sectioned for TEM. No method is really artifact free, but some artifacts are more easy to recognize than others. Knife induced artifacts are anisotripic (e.g. directional) in nature and can be more easily recognized (as artifacts) than artifacts caused by say, ion milling, which are isotropic in nature.
When I say "right kind of diamond knife", I am not suggesting that the SPI diamond knife could do something above and beyond what other diamond knives could do. What I am saying is that there is a process of selection of the optimum knife angle, because one might have to be prepared to use a knife with a fairly low (for materials science work) knife angle, rather than a more blunt angle, but with the downside is that it will wear out more quickly. That might make for great business for a diamond knife supplier, but it does tend to get expensive for the user who is not so experienced with this kind of sectioning.
Disclaimer: SPI Supplies offers diamond knives, both materials and life science, and we have done this kind of sectioning, as a service, for commercial clients for over thirty years.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Hi, The optics of forming an image is constrained by diffraction so that when the light comes together in a point it actually has a finite breadth. This is given by the numerical aperture of your objective and condenser (and the wavelength of light) and has a theoretical min of around 250 nm for green light. If you are using a high NA lens (say 1.3) and an equally good condenser, which must be oiled to the slide to get its full resolution, then you should be able to reach this limit. To the extent that your lenses are not that high NA, the limit will be larger. Vesicles smaller than the limit may be imaged (indeed, single microtubules can be imaged) but the diameter will read out on the image as being whatever your diffraction limit is. You can find a good discussion of this in Inoué's book on Video Microscopy, to name just one source.
As ever, Tobias
} } Hello all, } } I have a question about measuring small cellular structures at the light } microscope level using Argus 10 enhancement and measurement software. } } I am measuring vesicle diameters in living cells of the fungus Neurospora. } They measure about 350-400 nm in diameter using the Argus 10 software. } With cryofixed TEMs of the same cell type, the only vesicles with the same } distribution as at the LM level measure 150 nm diameter and there are no } structures at 350-400 nm. } } All my calibrations of both the LM and TEM have been checked and } double-checked. } } It is my understanding that video-enhanced LM can amplify signal (or sizes) } so that structures below limit of LM resolution can be detected (eg, } imaging of individual microtubules with DIC/VELM). I am wondering if this } is the source of this discrepancy that it see and if there are references } that describe this. } } Thanks so much,,,,,, } Robby Roberson } } } *************************************** } Robert W. Roberson, PhD } Department of Plant Biology } Molecular and Cellular Biology Program } Arizona State University } Tempe, AZ 85282 } Office/Lab 480-965-8618
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Alan J. Kruger } Sent: Friday, June 8, 2001 1:05 PM } To: Microscopy } Subject: LM: Formula for calculating dept of focus } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone lead me to a reference or the formula to calculate dept of } focus on light microscopes? } } Al Kruger } USDA Meat Animal Research Center } } }
A few days ago someone asked about sources for used and renovated microscopy equipment. Just today I received a leaflet from Microscopy Laboratories, (P.O. Box 338, Red Bank, NJ 07701, email:micro-at-mail.superlink.net; Tel: 732-747-6228) that lists a wide variety of items of reconditioned and guaranteed equipment they are offering for sale. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Dear All, I have been contacted by someone wishing urgently to obtain an image of skin with psoriasis for a science exhibition aimed at children. They specified an SEM image but possibly TEM or LM would do. We have nothing along these lines in our lab - is anyone out there in the wide world of microscopy able and willing to provide such an image?
Best regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
} Can anyone lead me to a reference or the formula to calculate dept of } focus on light microscopes?
Al,
"Photomicrography. A comprehensive treatise" by Roger P. Loveland John Wiley & Sons, 1970, contains an appendix on calculating depth of field. For compound microscopes Loveland gives several formulae, which produce similar but not identical results in his worked examples.
I can fax it to you if you can't find a copy locally.
Regards
Stephen Edgar
Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
This thread may never finish. Fred Monson wrote to me making the point that he thought that the original request (which I have appended last) related to a carbon sponge coated with Ta. Fred also had and so he appended an article in pdf format, which deals with such material. There also is what appears to be an SEM image of a thick section of this very open material. It seems that it had been sectioned, presumably with a diamond or TC knife. It does however, not show any cell inclusions, though the article says that purpose of the matrix is the growths of T-cells. Plastic infiltrated Ta coated C sponge with cell inclusions "may" section. The original correspondence referred to a Ta sponge coated with carbon with cell inclusions and that would be a whole lot more difficult. The publication is: Contact: Mark J. Pykett, Cytomatrix, (781) 939-0995, mpykett-at-cytomatrix.com I think that we could have a delightful time with a hundred protagonist writing to the unsuspecting "Contact" Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, June 12, 2001 12:38 AM, Alan Bright [SMTP:bright-at-dial.pipex.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would like to be of some assistance on the subject of a solution } to:-'There is no simple solution to the preparation of very hard and very } soft materials in one specimen.' } } As our Bright/Hacker OTF/AS/D/LT cryostat is able to section a hard and soft } specimen incorporated into one specimen. If this is of interest I would be } pleased to receive sample and return our results to those who have a } problem. } } Best Regards } } Alan Bright } } Bright Instrument Co.Ltd. } St Margaret's Way } Huntingdon } Cambridgeshire } PE29 6EU } England } } Tel No:+44 (0)1480 454528 } Fax No:+44 (0)1480 456031 } Email: AlanBright-at-brightinstruments.com } Web Site: www.brightinstruments.com } } } -----Original Message----- } } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] } Sent: 07 June 2001 23:26 } To: MICROSCOPY BB } Subject: Sectioning Ta } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } } ==================================================== } Tantalum is very hard and tough, so I doubt that it will section with either } } diamond or tungsten carbide knives. Preparing the material like a geological } } section by grinding is a possibility, but not a good one. Chances are that } the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in } plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a } diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } ===================================================== } Actually Ta **can** be thin sectioned if it is done by someone with the } proper experience and who is using the right kind of diamond knife. We have } offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for } some number of years; see URL } http://www.2spi.com/catalog/standards/aem.html } } The Ta system that prompted the original posting was thicker, however, it is } also porous, and if properly infiltrated with resin, and assuming the } porosity is above some point, in principle, at least, there is no reason why } it could not be thin sectioned for TEM. No method is really artifact free, } but some artifacts are more easy to recognize than others. Knife induced } artifacts are anisotripic (e.g. directional) in nature and can be more } easily recognized (as artifacts) than artifacts caused by say, ion milling, } which are isotropic in nature. } } When I say "right kind of diamond knife", I am not suggesting that the SPI } diamond knife could do something above and beyond what other diamond knives } could do. What I am saying is that there is a process of selection of the } optimum knife angle, because one might have to be prepared to use a knife } with a fairly low (for materials science work) knife angle, rather than a } more blunt angle, but with the downside is that it will wear out more } quickly. That might make for great business for a diamond knife supplier, } but it does tend to get expensive for the user who is not so experienced } with this kind of sectioning. } } Disclaimer: SPI Supplies offers diamond knives, both materials and life } science, and we have done this kind of sectioning, as a service, for } commercial clients for over thirty years. } } Chuck } original request was posted on 6 June 2001
I am posting this for a college, Lauri Wyner, in the Pathology Core. I have no idea how to section this stuff...!
Thanks! Maria Ericsson
My question is: I have a carbon coated porous tantalum matrix approximately 1 cm in circumference and 2 mm thick. This matrix has fixed cells adhered to the surface and throughout. I would like to embed this, make slides and stain for H&E to confirm the presences of cells as well as immunohistochemistry to characterize them. I am looking for suggestions on how I can section this matrix while maintaining its overall structure. Any information would be greatly appreciated.
POSTDOCTORAL POSITIONS IN NANOTECHNOLOGY AND BIOPHYSICS AT THE DEPARTMENT OF PHYSICS, UCLA
We are seeking outstanding candidates for postdoctoral research. Both spectroscopy/microscopy techniques and a strategic synthetic approach are used for materials discovery and studying the physics these novel materials. DNA/gold particles are used to probe the melting and duplex formation of DNA in confined geometries. Cryoelectron microscopy is used to study the structure and function of macromolecular assemblies at the interface between biological machines and nanotechnology.
Current Research Areas: Carbon Nanotubes Molecular biophysics of DNA and proteins Structure and function of macromolecular assemblies
The compensation package is competitive, and medical benefits are included. Interested person should send cv and three letters of recommendation to: Professor Ching-Hwa Kiang Department of Physics & Astronomy 6-130 Knudsen Hall University of California Los Angeles, CA 90095-1547 chkiang-at-physics.ucla.edu
----------------------------------------------------------------- Dr. Ching-Hwa Kiang 6-130 Knudsen Department of Physics Phone: (310) 206-0563 University of California Fax: (310) 825-5734 Los Angeles, CA 90095-1547 chkiang-at-physics.ucla.edu http://www.physics.ucla.edu/people/faculty_members/kiang -----------------------------------------------------------------
A fellow here in the department has the interest in knowing by which mean an FEG W-crystal needle is cut. I only know that in the case of Schottky emitter, the submicron needle is cut in {100} direction and coated with ZrO2 to reduce the work function. But I have no knowledge in the manufacturing process of the tip (FIB?). Anybody who knows please shed some lights. Thanks in advance.
********************************* Chaoying Ni Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
I have a question as to technique or method regarding the quantification of Au/Sn in a "solder bump" application for an InP microcircuit. Essentially what I've been asked to do is provide the weight % of each element after this "solder" has been melted and re-flowed. The samples are cylindrical in shape with an x-section of 65 uM and a thickness of 34 uM. The Au/Sn portion sits atop this structure and is a thickness of ~ 2 uM.
My question is, how can I quantify, in Wt. %, the ratios of the two metals? I have EDX at my disposal but I am uncertain that I will be able to accurately quantify this alloy without a standard. One other issue is that the underlying metallization that primarily constructs this "bump" is also Au for the next 32 uM.
Any insight into this problem would be greatly appreciated by myself and my colleagues.
Peter Tomic Analytical Services Group Anadigics, Inc. 35 Technology Drive Warren, New Jersey U.S.A. 07090
An excellent Philips CM-10 transmission electron microscope is being offered for sale by Michigan State University. It is 1987 vintage and has been on service contract since new. We can demo this instrument at any time. Please contact Dr. Xudong Fan with questions, fanx-at-msu.edu, 517-353-4525.
Stanley L. Flegler, Acting Director Center for Advanced Microscopy
Chaoying Ni , I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Ken Converse owner Quality Images Delta, PA
Chaoying Ni wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello lister, } } A fellow here in the department has the interest in knowing by which mean } an FEG W-crystal needle is cut. I only know that in the case of Schottky } emitter, the submicron needle is cut in {100} direction and coated with } ZrO2 to reduce the work function. But I have no knowledge in the } manufacturing process of the tip (FIB?). Anybody who knows please shed } some lights. Thanks in advance. } } ********************************* } Chaoying Ni } Electron Microscopy Laboratory } Materials Science and Engineering } College of Engineering } University of Delaware } Newark, DE 19716 } ********************************* } } } }
I was cleaning today and I have a complete ready-to-plug wehnelt assembly for a JEOL JSM-5200 SEM to give away. We inherited this piece and it is just a bit different from the unit in our 5400, so please only ask if you really have a 5200 as it probably won't fit other models. Claim goes to the first school or non-profit that requests it, otherwise others.
Contact me if interested. Be willing to pay postage. ($10???)
Dale Callaham +++++++++++++++++ Dale A. Callaham Senior Microscopist, Stuck Jar Lid Opener, Odd Job Bear Central Microscopy Facility Morrill 4 North, Room 1 639 North Pleasant Street The University of Massachusetts Amherst, MA 01003-9278 ------------------------- Phone 413-545-3751 FAX 413-545-3243 email dac-at-bio.umass.edu http://www.bio.umass.edu/microscopy
After more than 20 years of faithful service at IBM Research we will be retiring our Cameca SX50 electron microprobe. The machine has three wavelength dispersive detectors as well as one energy dispersive detector. The machine is operational except for the computer system, which is a PDP-11. The machine has been dormant, but under vacuum for roughly 3 years. If anyone is interested in purchasing the machine, please send your offers to :
Dr Andrew J. Kellock Ion Beams Lab Almaden Research Center (408) 927 2353 kellock-at-almaden.ibm.com
We have an old Zeiss Photomicroscope II at our lab, mainly in wery good condition, but there are electronic problems with the automatic camera system. There are some faults in the large electronic box placed in the microscope table. We have possibilities to repair the microscope here at the university, but unfortunately we have only been able to get a copy of the "Gesamtschaltplan zum Photomicroscope III" (the electronic diagram), but that is entirely different. In order to repair it we need the original electronic diagram (or a copy of it) for the Photomicroscope II. The Carl Zeiss Factory could not help us. If someone could send us a copy, we would highly appreciate it.
Sincerely:
Morten Motzfeldt Laane Professor of Biology (electron-microscopy) Biological Institute P.O.Box 1066, 0316 Blindern, Oslo, Norway
Dear Sir or Madame, I have been asked by my employer to investigate setting my laboratory up to perform what was called "hot-stage microscopy". I understand this to be the ability to observe the melting of (in this case) a polymer. I was hoping that your organization could point me in the right direction. Any information you could give me would be greatly appreciated.
I am looking for a colouring agent for oil (emollients) which needs to applied to human skin. If we apply the oil as such we do not have enough contrast for measurements. This colouring agent needs to be skin friendly, leave no stain on the skin and it must not change the viscosity of the oil.
Any information would be very helpful and appreciated.
Thanks,
Nancy Meijer
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Below is the result of your feedback form. It was submitted by (lilaeloise-at-cs.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, June 12, 2001 at 16:14:22 ---------------------------------------------------------------------------
Email: lilaeloise-at-cs.com Name: Lila Allen
Organization: Sunrise Hospital
Education: Graduate College
Location: Nevada
Question: I've been looking for information regarding the differences between laser confocal microscope and digital confocal microscopes, including the advantages/disadvantages and uses of each. Thank you, Lila
Thanks to all who replied to my request for references to images of osmolarity problems. The book below is a wonderful resource for images of all kinds of treatments ranging from the effects of various fixation protocols to analysis of digital images. I don't know how this book escaped my notice when it came out, but I'm buying it, and heartily recommend it!
} Biomedical Electron Microscopy by Maunsbach A B & Afzelius } B A (1999) has several pages of illustrations of shrinkage } etc. }
} } Hi, All- } } } } Does anyone have a reference for or some transmission electron micrographs } } that illustrate the effects of osmolarity of buffers on animal cells? I } } would like to show students on tour of our facility shrinkage and swelling } } of cells and organelles. I have other wonderful artifacts to show, } } including one of my first sections with chatter so bad it looks like } } mini-blinds... } } } } Mahalo! } } Tina } } } } http://www.pbrc.hawaii.edu/bemf/microangela } } **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hello, } From time to time I need to scan some negatives with flat bed scanner (Umax PowerLook II). Sometimes there is a problem with Newton rings in the scanned image. Does anybody know some tricks how to solve this problem? Best regards Oldrich +-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of Electron Microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-2-4752399 Fax: +420-2-4715743 WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm
Emispec Systems, Inc., a rapidly growing company that creates software for electron microscopy, is looking for a domestic technical sales engineer. We are looking for someone with strong interpersonal skills and experience with customer interactions.
Some of the job requirements will include:
-Strong technical knowledge of products. -Ability to give product demonstrations at tradeshows and at customer sites. -Providing product information and quotations. -Defining potential customers. -Qualifying leads and determining customer needs. -Adding/updating contact information in the sales database. -Strong customer support throughout the purchasing cycle and after. -Handling initial customer service requests, and coordinating with the service department to see that they are handled appropriately. -Extensive travel.
The ideal candidate should have a college degree. Sales experience and/or experience in electron microscopy is a plus. Salary is commensurate with experience.
Emispec Systems, Inc. is an equal opportunity employer.
Resumes and salary requirements should be sent to: Human Resources Emispec Systems, Inc. 2050 S. Cottonwood Drive Tempe, AZ 85282 Attn: Technical Sales
Resumes will also be accepted via email to: hr-at-emispec.com.
Below is the result of your feedback form. It was submitted by (ggauss-at-dynacocorp.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, June 12, 2001 at 18:12:49 ---------------------------------------------------------------------------
Email: ggauss-at-dynacocorp.com Name: Gordon Gauss
Organization: Dynaco Corporation
Education: Undergraduate College
Location: Tempe, Arizona
Question: We recently received a microscope with no manual. It is an aus JENA Jenavert. It is in excellent condition. We believe it was East German made. Is anyone familiar with this microscope and how can we obtain a manual in either german or english. Thank you...
If Newton rings are the same as Moire patterns (constructive/destructive interference lines)....
Many scanner drivers have a "de-screen" option or the like. My Microtek system seems to do a decent job of removing the lines. Also, you might try scanning at a much higher dpi (dots per inch ...or cm), then lightly average pixels and reduce in array size to meet needs.
Woody White McDermott Technology, Inc
} -------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hello, } } From time to time I need to scan some negatives with flat } bed scanner } (Umax PowerLook II). Sometimes there is a problem with Newton } rings in } the scanned image. Does anybody know some tricks how to solve this } problem? } Best regards Oldrich } +-----------------------------------+ } Oldrich Benada } Acad. Sci. CR } Institute of Microbiology } Laboratory of Electron Microscopy } Videnska 1083 } CZ - 142 20 Prague 4 - Krc } Czech Republic } +------------------------------------+ } Phone: +420-2-4752399 } Fax: +420-2-4715743 } WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm } } }
I run a research based service lab at Ohio State University and I've been asked to find out what other universities charge their internal customers for the use of TEM, SEM, confocal microscopy and technical assistance. I'm particularly interested in biological labs at large state universities, big 10 universities, and Ohio universities. Please respond directly to me. Thanks. Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
The Newton rings are interference fringes due to the small air gap when the film not quite touches the glass. The way to get around the problem is to raise the film off of the glass just slightly. Most of the better quality scanners provide holders, although not TEM sized one. Make your own from two pieces of a file folder cut to the appropriate size and held together at one edge with tape. There is no problem with a defocus of the image.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Oldrich Benada [mailto:benada-at-biomed.cas.cz] Sent: Wednesday, June 13, 2001 2:45 AM To: microscopy-at-sparc5.microscopy.com
Hello, } From time to time I need to scan some negatives with flat bed scanner (Umax PowerLook II). Sometimes there is a problem with Newton rings in the scanned image. Does anybody know some tricks how to solve this problem? Best regards Oldrich +-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of Electron Microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-2-4752399 Fax: +420-2-4715743 WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm
Try speaking to Hitachi in Gaithersburg, MD. Their Applications Lab. should have that info. at their fingertips.
Peter Tomic Anadigics
-----Original Message----- } From: Ken Converse [mailto:qualityimages-at-netrax.net] Sent: Tuesday, June 12, 2001 4:39 PM To: Chaoying Ni; MSA, listserver
Chaoying Ni , I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Ken Converse owner Quality Images Delta, PA
Chaoying Ni wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello lister, } } A fellow here in the department has the interest in knowing by which mean } an FEG W-crystal needle is cut. I only know that in the case of Schottky } emitter, the submicron needle is cut in {100} direction and coated with } ZrO2 to reduce the work function. But I have no knowledge in the } manufacturing process of the tip (FIB?). Anybody who knows please shed } some lights. Thanks in advance. } } ********************************* } Chaoying Ni } Electron Microscopy Laboratory } Materials Science and Engineering } College of Engineering } University of Delaware } Newark, DE 19716 } ********************************* } } } }
I am looking for a LKB Nova ultramicrotome. I presently have a Reichert Ultracut E which works absolutely fine, its just that I'm really a fan of LKB ultramicrotomes (the last one was a LKB III which is not usable anymore). I have used LKB's for over 20 yrs and I would prefer to go back to that brand. Does anyone out there want to swap or sell one?
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642 716-275-1954
We have been looking at melting and crystallization of polymers under hot stages for twenty years. We have always found Mettler-Toledo Hot Stages very good, and have bought three of them.
For information, go to:
http://www.mt.com
then click on "Products" in the bar at top;
in the window 2.Product Search type "stage";
in the next window, under the text "Thermal Data - FP82HT/FP84HT"
click on the little button "English"
which will take you to a screen where you can request more information;
(that's all one line, in case the e-mail server has chopped it in two.)
** Disclaimer ** we have no commercial interest in M-T, simply we've found them very good and helpful.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
On Tue, 12 Jun 2001 Dalton.Pierson-at-sealedair.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Sir or Madame, } I have been asked by my employer to investigate setting my laboratory up to } perform what was called "hot-stage microscopy". I understand this to be the } ability to observe the melting of (in this case) a polymer. I was hoping } that your organization could point me in the right direction. Any } information you could give me would be greatly appreciated. } } Thank you, } Dalton Pierson } }
You do not indicate whether the objective is to measure the total amount of the two metals in the assembly as a whole, or whether you want to measure their proportions in each of the different areas: the tin solder, the intermetallic layers and so on. You also don't indicate the purpose to which the results are to be put. To some extent the answer depends on knowing this, as a biased but highly repeatable measure of something is often useful for internal purposes.
Someone with specific experience in this industry might have a more specific reply but in general, the problem is that the same elements are shared by very different phases in close proximity to one another. For the Au/Sn intermetallic compounds which have quite specific compositions, the absorption /enhancement effects will be different than for a low-concentration solid solution of either of the metals in the other. Without a standard comprised of a known intermetallic phase, the results could be consistent (that is, you might be able to use the measure as a means of process control) but not very accurate (as compared with the absolute amounts present). Another frequently overlooked problem with quantifying the results of e-probe analysis of a multiphase alloy is that the size distribution of imiscible but intimately mixed phases may change due to process variables (e.g. thermal history) even though the total proportion of the metals remains the same. The micro-environment (matrix effect) experienced by an emitted x-ray will depend on its probability of passing through the same material or a different phase on its way to reaching the detector. Obviously this can be affected by the grainsize of a eutectic mixture.
You haven't indicated whether the sample is to be sectioned into a planar surface for analysis or examined as is. Another frequently overlooked problem is that of x-rays emitted through secondary interactions with structures that are not illuminated by the beam but which are "visible" within the detector collimator's view. A planar cross section presents the least chance for interference of this kind. On the other hand, a gold-metallized surface projection that resides next to your solder structure could be a source of enhanced Au-m x-rays produced through secondary fluorescence by the Sn-l x-rays under the beam. (even though secondary emmission is not a very efficient process, the number of Sn x-rays fluorescing the adjacent gold is much higher than the number that happen to be emitted in the direction of the X-ray detector. Meanwhile the X-ray detector is looking at the whole scene indiscriminately.
John Twilley
Peter Tomic wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Folks; } } I have a question as to technique or method regarding the quantification of } Au/Sn in a "solder bump" application for an InP microcircuit. Essentially } what I've been asked to do is provide the weight % of each element after } this "solder" has been melted and re-flowed. The samples are cylindrical in } shape with an x-section of 65 uM and a thickness of 34 uM. The Au/Sn } portion sits atop this structure and is a thickness of ~ 2 uM. } } My question is, how can I quantify, in Wt. %, the ratios of the two metals? } I have EDX at my disposal but I am uncertain that I will be able to } accurately quantify this alloy without a standard. One other issue is that } the underlying metallization that primarily constructs this "bump" is also } Au for the next 32 uM. } } Any insight into this problem would be greatly appreciated by myself and my } colleagues. } } Peter Tomic } Analytical Services Group } Anadigics, Inc. } 35 Technology Drive } Warren, New Jersey } U.S.A. 07090 } } } }
Hi, I've found two things that helped eliminate this problem. If you place the negative directly onto the scanner bed, use an anti-static brush to wipe both the bed and the negative first. The second solution was to modify a negative holder. I work mostly with TEM sized negatives and we just cut out the back (solid) part of a TEM negative holder, then we place the negative into the holder which keeps it from contacting the scanner bed directly. If your working with a different size negative you could fashion a holder from anything, even a piece of poster board should work. I think the fringes are caused by a build up of static between the scanner bed and the negative. I have also found that if you go to adjust contrast in print shop and adjust the contrast through the entire range from too light to too dark you can pick up a fringe before you take the negative off the scanner. That eliminates having to go back and re-do a picture because the fringe wasn't real apparent when you scanned it. Hope this works for you...it has made my life a lot easier. Dorrance
} ---------- } From: Oldrich Benada } Sent: Tuesday, June 12, 2001 11:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Unwanted Newton rings } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } From time to time I need to scan some negatives with flat bed scanner } (Umax PowerLook II). Sometimes there is a problem with Newton rings in } the scanned image. Does anybody know some tricks how to solve this } problem? } Best regards Oldrich } +-----------------------------------+ } Oldrich Benada } Acad. Sci. CR } Institute of Microbiology } Laboratory of Electron Microscopy } Videnska 1083 } CZ - 142 20 Prague 4 - Krc } Czech Republic } +------------------------------------+ } Phone: +420-2-4752399 } Fax: +420-2-4715743 } WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm } } } } }
Job Master, a complete, user friendly Windows based software package, can manage and control your operation from sales quote to shipment.
For one week only, Job Master, normally $2,495.00, is on sale for a total price of $1,495.00. In order for you to receive this $1,000.00 savings we must have your order by June 22th.
Job Master is designed specifically for small to medium sized manufacturers, and costs many thousands of dollars less than any other even remotely comparable software package.
Following is a list of features. If you have any questions, would like to discuss the package further, or if you would like to obtain our Web site address for a total walk through of the program, please call me directly at (661)254-9926(please do not E Mail me back. I may not get your message if you simply hit "Reply" and respond to this message via return E Mail).
By way of background, we are a software company, which for some years has specialized in the development of custom software, primarily for small to medium sized manufacturers. Job Master is a distillation of over a million and a half dollars of software we have developed to control and manage the production of our manufacturing clients.
Job Master contains the following features:
1. QUOTATION MODULE. In this module, quotes are developed, modified, and produced for sending to your client. A history is kept of all quotes for future reference, or modification for other clients. All quotations and revisions are "auto numbered," including versions. The quotes section allows for the entry of parts/processes, and costing of each, including materials, labor, markup, and taxes. Inventory status can be accessed from this section for reference.
2. SALES ORDER. Once a quotation is accepted, the final quotation information can be transformed into a Sales Order for your client's signature on a "point and click" basis. The Sales Order can be modified and re issued if necessary. A history if kept of all Sales Orders for future reference, or modification for other clients. All sales orders and revisions are "auto numbered," including versions. Inventory status can be accessed from this section for reference.
3. CUSTOMER LETTERS can be created from the Quotation and Sales Order sections.
4. SHOP TRAVELER/WORK ORDER. Once a Sales Order is accepted, the sales order information can be transformed into a shop traveler/work order on a "point and click" basis. Each item on the Sales Order becomes a shop traveler/work order, with each step of production of the item then listed on the traveler/work order. Each such traveler/work order is tied back into the Sales Order. The shop traveler/work order allows for the entry of line items, and notes on each line item. The shop traveler/work order contains a "notes" section. The Shop traveler/work order allows for the storing or attachment of drawings to the traveler/work order. The shop traveler/work order also contains a "drop down," from which standard processes can be selected for inclusion on the shop traveler/work order. The shop traveler/work order numbers progress in order of production sequence, and re numbers them if new steps are added. The shop traveler/work order allows for change orders or revisions, and numbers changes in sequence of he original shop traveler/work order number; i.e., 100, 100-1, 100-2, etc. All shop traveler/work orders and related revisions are retained in memory for future reference. The shop traveler/work order is bar coded for tracking of production step by step, and production of ongoing client status reports. Bar coding includes the ability for an employee to "swipe" their own ID bar code for recording in the system as to who upgraded what step. The shop traveler/work order function also allows for manual update of production status. The shop traveler/work order allows for quality control sign off, and the final production of certifications, either from a "canned" list, or hand typed in on a case by case basis.
5. INVENTORY. The application includes an inventory section, which allows operations to check materials inventory in and out. The inventory section allows for the comparison of inventory received against a P.O., and produces an "overage/underage" report of inventory received as compared against the P.O. The inventory section allows for the setting of minimum (re-order now!) and maximum inventory amounts, and produces reports showing what inventory needs to be ordered, as well as inventory that is at or above the maximum set to have in house. The inventory section also tracks "partially shipped" orders, which are tied in to the shipping function. This section shows how much completed product under a particular order has been actually shipped to a client, and how much remains to be shipped. The balance is adjusted as shipments are made.
6. REQUEST FOR PURCHASE. The application allows operators to produce a Request For Purchase for accounting for any inventory items, which need to be ordered. Inventory items have a drop down of approved vendors for each item.
7. REQUEST FOR BID. The application allows operators to produce a Request For Bid for accounting to send to Vendors for any inventory items, which need to be ordered. Inventory items have a drop down of approved vendors for each item to which Requests For Bid can be sent.
8. INVOICE. The application produces an invoice/invoice detail for all completed items ready to be billed/shipped to clients.
9. PRODUCTION OUTPUT STATUS. The application produces a date range selectable report on how much product, and the value of the product, which was completed during a selected date range. The application also produces a report on how many orders, and the value of those orders, which remain to be completed during a selected date range.
10. The application produces SHIPPING DOCUMENTS as per selected shippers, and produces a PACKING SLIP.
11. The application has a "FIND" FUNCTION in selected sections, allowing for searches by customer name, work order number, etc.
12. The application has "AUTO FILL;" i.e., when an operator starts to type in a name, number, etc. all related information auto fills after the first few letters or numbers are typed in.
Job Master is currently being sold in the marketplace for $2,495.00 per package. However, if we receive your order by June 22th, your total price will be $1,495.00.
Again, if you have any questions at all, or would like to place your order, please call me on my direct line, (661) 254-9926. Thank you!
You have received this newsletter because you signed up for updates on our tracking software. If you want to unsubscribe from this newsletter, please send a reply email with "REMOVE" in the subject line.
Listers: Here's info about another source of used instruments.
- - - - - - - - - - - -
} From: "A. Greene" {ablue-at-io.com} } To: {bigelow-at-umich.edu} } Subject: Source } Date: Mon, 11 Jun 2001 20:05:13 -0500 } X-Priority: 3 } Status: } } Hello Professor Bigelow, } } Here is another source, as well. I have been in the instrument } business for over 30 years, used to be with Philips and then at the } University of Illinois for 10 years. Now, for the past eight years } have had my own little business in Texas. I probably have more } parts for older Philips microscopes than Philips. Also, I sell } reconditioned, used electron microscopes of most brands. } } Regards, } } Alex Greene } SCIENTIFIC INSTRUMENTATION SERVICES, INC. } PMB-499, 1807 West Slaughter Lane, Number 200 } Austin, Texas 78748-6200 } Phone 512/282-5507 FAX 512/280-0702 } } Sustaining Member - MICROSCOPY SOCIETY OF AMERICA
-- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
We have a graduate student who is using a fluorescent antibody labelling technique on a neuronal cell culture plate. Briefly, she fixes the cells in 4% paraformaldehyde, permeabilizes them with Triton-X and continues with the immunostaining. The cells stay on the plate (they can only grow on plates, not slides or coverslips). The question we have is how to coverslip them and then observe them with a fluorescent microscope. Will glycerol work? Can we simply put a drop of glycerol on the plate and add a coverslip or do we need a fluorescent mounting medium? I have limited experience with fluorescent microscopy, so I would appreciate any help that you can provide.
Thank you very much, Sandy Hancock
Laboratory for Neurotoxicity Studies VMRCVM VA Tech
I am using transparency for Xerox machine with "windows" cut in it in such manner, that it's slightly smaller than actual negative's size. In my particular case I find that rings happens between negative and scanners bed. So, I put transparency on the bed and than negatives (4 per transparency sheet) in the way, when negatives situated across the "windows". In my particular case, transparency presence does not affect scanned image quality. You may put second sheet over the negatives if necessary.
Good luck!
Sergey.
At 10:00 AM 6/13/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Recently, there was a question about TEM and osmolarity. I have now a question about SEM and osmolarity. Has anyone have some references about this topic ? We are studying the canine uterus. We have some problems concerning (we think !) the osmolarity & the epithelial cells of the uterus. Any comment would be appreciated !
Photobleaching, or fading, causes loss of fluorescent signal in part due to photodynamic events which involve the interaction of the fluorophore with light energy and oxygen. Fluorescent labels have a characteristic life span, for instance, an average fluorescein molecule will emit 30k to 40k photons during its photo-chemical life span. Higher intensity light excites the fluorophor to emit a greater yield of photons and the service life of the fluorescent label is proportionally reduced. For this reason, it is generally a good idea to add some form of bleaching protection (anti-fading reagent) to a glycerol-based mounting media for fluorescent microscopy. P-phenylenediamine (Sigma Chemical Co.) used in a concentration of 0.1% in [10% phosphate buffered saline/90% glycerol] is good. Diazabicyclo-octane or n-propyl-gallate (~0.1-0.2% in 10% PBS/Glycerol are decent alternatives. The disadvantage to this approach is that the coverslip must be sealed (nail varnish) in order to fix the coverslip permanently. Vectrashield (Vector Co.) is a propriety product for fluorescent microscopy which seems to work quite well with the added advantages of convenience and greater permanence. One other consideration is the degree to which your fixation protocol affects the antigenicity of your target-a short (10 min/10% buffered formalin) is a good place to start-some antigens seem to be very sensitive to the cross-linking effects of aldehyde fixation, acetone or ethanol/methanol fixation may prove helpful if this is the case. If the target is a membrane protein one wants to be cautious with the Triton-X. Residual Triton-X could also interfere with the antigen-antibody interaction in a sensitive system. Acetone also permeablizes cell membranes, and if I recall correctly is a favorable fixative for immunofluoresent studies of the cytoskeleton. Culturing cells on slides or coverslips may not be an issue as long as you can get the culure plates into the scope. However, I believe theromonox (sp?) coverslips are designed to allow cell adhesion. Some workers coat slides and/or cover slips with poly-l-lysine for cell adhesion. Coverslipping is important to allow the optical system of the microscope to function at full potential. This is less of an issue with inverted microscopes designed to accomodate tissue culture vessels. Regards, Karl Garsha
You can de-stain both thick and thin section w/ 0.2 M EDTA. Only the time differs. One caveat, if the section has been irradiated, i.e. in the TEM then this will much less effective.
Good luck
Richard Cole Research Scientist III Director: Advanced Light Microscopy Core Unit Wadsworth Center P.O. Box 509 Albany N.Y. 12201-0509 518-474-7048 Phone 518-486-4901 Fax
-----Original Message----- } From: "tbargar-at-unmc.edu"-at-sparc5.microscopy.com [mailto:"tbargar-at-unmc.edu"-at-sparc5.microscopy.com] Sent: Wednesday, June 13, 2001 4:10 PM To: Microscopy-at-sparc5.microscopy.com
Hello;
Is it possible to destain a thin section? After having stained with uranyl acetate and lead citrate. All help appreciated, thanks.
At the University of Rochester Electron Microscope Research Core, we charge $75.00/hour with an Electron Microscopist running & photographing using our Hitachi 7100 TEM, without the EM person, a trained researcher or grad student must be trained on its use and the rate becomes $50.00/hour.
Our processing and preparation of tissue is at $75.00/hour and that only counts the hands-on times. For instance, usually it could take 30 minutes to do 1 micron sections and thin section the same block. We bill for .5hr=$37.50 to cut one block, .25 hr=$18.75 if no screening 1 micron sections are needed.
Most investigators have us embedd 2-6 blocks of tissue, etc. We cut a representative block, review the 1 micron with them if necessary and then proceed with the thin-sectioning. Staining is usually .25hr...etc, If the investigator wanted more than one block cut and thin sectioned(3-5 grids), they would all be stained together using a Hiroka staining kit(holds up to 40 grids) and therefore would only be charged the .25hr rate. However, the methods and materials you use and speed in which they are performed can vary from one lab to another.
We only billed $35,000 last year and that doesn't begin to cover salaries. So the University of Rochester has to subsidize the EM Core. However, the Core(s) is a big marketing tool for those researchers which are actively being recruited. Many Molecular Science researchers utilize the EM Core for ImmunoEM procedures. In fact, the majority of my work is performing pre- and postembedding immunolabeling for EM. The average bill for this varies from $00-1,000.
Hope this helps you.
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Kathy Wolken [mailto:wolken.1-at-osu.edu] Sent: Wednesday, June 13, 2001 9:15 AM To: microscopy-at-sparc5.microscopy.com
I run a research based service lab at Ohio State University and I've been asked to find out what other universities charge their internal customers for the use of TEM, SEM, confocal microscopy and technical assistance. I'm particularly interested in biological labs at large state universities, big 10 universities, and Ohio universities. Please respond directly to me. Thanks. Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
As a general rule, I am less than thrilled about using anti-fade agents for fixed cells unless absolutely necessary for the reasons explained below. Live cell work may require anti-oxidants but I am less familiar with this application. I think it is better to use more stable fluorochromes (e.g., Alexa 488 instead of FITC)
PPD is a skin sensitizer, photosensitive, and Giloh & Sedat (1982) Science 217:1252 says it has no effect on rhodamine fading. I believe they actually showed storage of rhodamine specimens in 5% n-propyl gallate (NPG) in glycerol for 2-3 days decreased fluorescence (suggest rinsing slides for storage (what a pain!). Krienk et al., 1989 J. Immuno. Methods 117:91 compared DABCO, PPD and NPG and concluded DABCO the least effect. Valnes & Brandtzaeg (1985) J Histochem Cytochem 33:755 also said DABCO not as effective as PPD or NPG but also say NPG and PPD only good if slides made recently and examined promptly - recommended re-mounting in PVA without quencher if you were going to store. I have tried Vectashield and other commercial anti-fade compounds with ok results but it is hard to say exactly how much they helped. I don't think there are any systematic published studies examining if they work equally well with all fluorochromes or if they hurt the sample with storage.
Be cautious about using FITC in any PBS buffered mounting medium since it is much more fluorescent at alkaline (pH 8.6) than neutral (pH 7.0) pH's.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I'm trying to embed and section mouse cochleas that have been immunoreacted en bloc for BrdU with DAB as the chromagen. The soft tissues of the cochlea were separated from the bony structures, leaving a soft tissue spiral containing the organ of Corti.
During processing for Epon-Araldite for 5 micron sections for light microscopy (without osmium and using increasing concentrations of aqueous acetone as the dehydrant), it appears as if the chromagen was lost (no labeled cells were seen) and a purple/grey background was left behind. Whole mounts immunoreacted at the same time retained an amber coloring with dark brown-labeled nuclei.
Does anyone have any hints as to what happened here or what we can do in the future to retain the labeling throughout plastic processing. I've done this embedding in the past with the same type of tissue (but still in the bony labyrinth) and had no problems.
Thank you,
Jaclynn M. Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
Just an FYI- we have found thermanox to be autoflourescent.
Lara Muffley Dermatology Dept University of Washington Seattle, WA muffley-at-u.washington.edu
On Thu, 14 Jun 2001, Karl Garsha wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings Sandy, } } Photobleaching, or fading, causes loss of fluorescent signal in part due to } photodynamic events which involve the interaction of the fluorophore with } light energy and oxygen. } Fluorescent labels have a characteristic life span, for instance, an average } fluorescein molecule will emit 30k to 40k photons during its photo-chemical } life span. Higher intensity light excites the fluorophor to emit a greater } yield of photons and the service life of the fluorescent label is } proportionally reduced. } For this reason, it is generally a good idea to add some form of } bleaching protection (anti-fading reagent) to a glycerol-based mounting } media for fluorescent microscopy. P-phenylenediamine (Sigma Chemical Co.) } used in a concentration of 0.1% in [10% phosphate buffered saline/90% } glycerol] is good. Diazabicyclo-octane or n-propyl-gallate (~0.1-0.2% in } 10% PBS/Glycerol are decent alternatives. The disadvantage to this approach } is that the coverslip must be sealed (nail varnish) in order to fix the } coverslip permanently. } Vectrashield (Vector Co.) is a propriety product for fluorescent } microscopy which seems to work quite well with the added advantages of } convenience and greater permanence. } One other consideration is the degree to which your fixation protocol } affects the antigenicity of your target-a short (10 min/10% buffered } formalin) is a good place to start-some antigens seem to be very sensitive } to the cross-linking effects of aldehyde fixation, acetone or } ethanol/methanol fixation may prove helpful if this is the case. If the } target is a membrane protein one wants to be cautious with the Triton-X. } Residual Triton-X could also interfere with the antigen-antibody interaction } in a sensitive system. Acetone also permeablizes cell membranes, and if I } recall correctly is a favorable fixative for immunofluoresent studies of the } cytoskeleton. } Culturing cells on slides or coverslips may not be an issue as long as } you can get the culure plates into the scope. However, I believe theromonox } (sp?) coverslips are designed to allow cell adhesion. Some workers coat } slides and/or cover slips with poly-l-lysine for cell adhesion. } Coverslipping is important to allow the optical system of the microscope to } function at full potential. This is less of an issue with inverted } microscopes designed to accomodate tissue culture vessels. } Regards, } Karl Garsha } } keg-at-uwm.edu } www.uwm.edu/~keg } } } } } } } } } } } }
I am in the market to purchase a new ultramicrotome. I am aware of the Leica and RMC ultramicrotomes. Are there other companies manufacturing ultramicrotomes today?
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help.
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, Maryland
Do you have filters somewhere in the recirculation lines? A lot of places have 2 barrel-type filters (I don't know the correct term - they look like tubing, not flat membranes) in-line to clean up anything that might grow. Change them every 6 months or when they start to look like they might become sentient.
I've been told that you should not put algicides in the water; the filters are probably the best way to go.
Tamara
On Thu, 14 Jun 2001, Haixin Xu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi lister, } } I have some problems with our EM chillers. The water in EM cooling } circulation is bad contaminated by algae, bacteria, maybe more } microorganisms. Does somebody here know some water cleaner for chillers of } EM? Appreciate for the help. } } Haixin Xu } } Biological Sciences } University of Maryland, Baltimore County } Baltimore, Maryland } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Below is the result of your feedback form. It was submitted by (Whunter-at-ushrl.ars.usda.gov) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, June 14, 2001 at 07:58:06 ---------------------------------------------------------------------------
Email: Whunter-at-ushrl.ars.usda.gov Name: Wayne Hunter
Organization: USDA { ARS
Education: Graduate College
Location: Ft. Pierce, FL, USA
Question: I am looking for a protocol to embed insects in Paraplast, and doing thick sections, Thank you, Sincerely, Wayne Hunter REsearch Entomologist
Growth of algae, bacteria etc may also be prevented/reduced by using black, intransparant tubes instead of white transparant ones. They were installed on my chiller for an ESEM 2020, after considerable problems. With best regards, Emond de Roever, Ondeo Nalco Europe Technical Centre, Oegstgeest, the Netherlands
"I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help."
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, Maryland
Haixin, I've found that using distilled or de-ionised water in the chiller in addition to the measures already suggested will also greatly reduce the growth of microorganisms by denying them nutrients.
Regards, Eric lachowski
------------ Dr Eric Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland Tel. +44 (0)1224 272934 Fax +44(0)1224 272921 e.lachowski-at-abdn.ac.uk
I am using an Epson 980. I find it satisfactory for both B&W and color prints and it is reasonably fast. It is also relatively inexpensive. Choice of paper grade/type is very important in determining available print quality.
Woody White McDermott technology, Inc
} -----Original Message----- } From: treese [mailto:treese-at-marinebio.mbl.edu] } Sent: Friday, June 15, 2001 1:00 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: printers } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } ------------------------------------------------------------- } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------- } ----------. } } } } } We are going to replace our old Epson InkJet printer which produces } nice B+W prints of micrographs, when it works. I haven't kept up } with Epson vs HP for electron microscopists and wondered if a clear } preference for one of these, or maybe another mfg or technology has } emerged. We want B+W quality and reliability, and are willing to pay } a bit more,and sacrifice speed and color quality...Thanks...Tom Reese }
I have seen a retrofit for Epson printers that looks quite interesting at
http://www.piezography.com/
I haven't actually seen it in action but it looks like it should be good for B/W prints. The advertising samples on the web site look very nice. Basically they replace the color cartridges with different densities of black ink. By allowing gray inks, they reduce the amount of dithering required. The product consists of the replacement cartridges and the revised printer driver software.
Has anyone actually tried this approach?
Cheers, Henk Colijn
At 12:00 AM 6/15/01 -0500, treese wrote:
} We are going to replace our old Epson InkJet printer which produces nice } B+W prints of micrographs, when it works. I haven't kept up with Epson vs } HP for electron microscopists and wondered if a clear preference for one } of these, or maybe another mfg or technology has emerged. We want B+W } quality and reliability, and are willing to pay a bit more,and sacrifice } speed and color quality...Thanks...Tom Reese
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Does anyone happen to know in what issue of microscopy Today the following information on sharpening W needles was found? (or how to make them) If so please send me a note at, MRoberson-at-dow,com
I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Michael Roberson (517)636-8656 SMX Analytical Sciences Midland, MI 48667
The question of avoiding algal growth in water chillers is discussed at length on p. 216 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html and http://www.pup.princeton.edu/titles/6484.html)
One important way of reducing algal growth is to exclude light from the system by keeping the reservior tank covered and by using opaque tubing. If this is not sufficient then we have had great success over many years by adding the algacide known as 'Chloramine-T' to the water in the amount of about 0.25 grams per liter. This stuff is available from most speciality chemical companies (Aldrich, Sigma, Polysciences) under the chemical name of N-chloro-p-toluensulphonamide. It is not highly soluble, and so is not likely to produce deposits in the system. We have not had any trouble with damage to any of our systems in which this material has been used. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
-----Original Message----- } From: Jensen, Karen Sent: Thursday, June 14, 2001 10:23 AM To: 'Kathy Wolken' Cc: 'microscopy-at-msa.microscopy.com'
Dear Kathy:
At the University of Rochester Electron Microscope Research Core, we charge $75.00/hour with an Electron Microscopist running & photographing using our Hitachi 7100 TEM, without the EM person, a trained researcher or grad student must be trained on its use and the rate becomes $50.00/hour.
Our processing and preparation of tissue is at $75.00/hour and that only counts the hands-on times. For instance, usually it could take 30 minutes to do 1 micron sections and thin section the same block. We bill for .5hr=$37.50 to cut one block, .25 hr=$18.75 if no screening 1 micron sections are needed.
Most investigators have us embedd 2-6 blocks of tissue, etc. We cut a representative block, review the 1 micron with them if necessary and then proceed with the thin-sectioning. Staining is usually .25hr...etc, If the investigator wanted more than one block cut and thin sectioned(3-5 grids), they would all be stained together using a Hiroka staining kit(holds up to 40 grids) and therefore would only be charged the .25hr rate. However, the methods and materials you use and speed in which they are performed can vary from one lab to another.
We only billed $35,000 last year and that doesn't begin to cover salaries. So the University of Rochester has to subsidize the EM Core. However, the Core(s) is a big marketing tool for those researchers which are actively being recruited. Many Molecular Science researchers utilize the EM Core for ImmunoEM procedures. In fact, the majority of my work is performing pre- and postembedding immunolabeling for EM. The average bill for this varies from $300-1,000.
Hope this helps you.
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Kathy Wolken [mailto:wolken.1-at-osu.edu] Sent: Wednesday, June 13, 2001 9:15 AM To: microscopy-at-sparc5.microscopy.com
I run a research based service lab at Ohio State University and I've been asked to find out what other universities charge their internal customers for the use of TEM, SEM, confocal microscopy and technical assistance. I'm particularly interested in biological labs at large state universities, big 10 universities, and Ohio universities. Please respond directly to me. Thanks. Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
you can purchase algicide from Fisher, VWR or other lab suppliers.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Haixin Xu {xu-at-gl.umbc.edu} 06/14 5:22 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi lister,
I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help.
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, Maryland
I don't remember which issue of MT, but the following two paragraphs are three items that I sent to the Listserver recently describing sample preparation of emitters.
1)(Ni) I'm trying to remember back to grad school, but I'm pretty sure that it was a 10%HCl solution in water. I used it for Ni FIM samples. I'll look it up in my dissertation and see if I have it. Hren's book or Bowkett and Smith's book on FIM had the reference for Ni. I do remember that the solution worked better after it was used some. It took on a greenish tinge. So what I started doing with a fresh batch, was taking a little crystalline NiCl (green powder) a putting just a bit into solution to get the color just right. I can't remember if I used ac of dc.
2)(W) I used to prepare W FIM emitter tips with a 5% NaOH solution and I think the voltage was around 5 to 10 volts ac. When I used to electropolish, the FIM solutions and conditions were frequently similar to the jet polishing solutions.
3)(Fe & floating layer method) You can use Field Ion Microscopy sample prep techniques to prepare very sharp needles of the wires. Basically, you can dip them in a beaker and electropolish them. The liquid/air interface will preferentially polish them. You need to move the interface up and down the sample by moving either the sample or liquid. If the wires are long enough, you can float electrolyte on top of CCl4 layer and you can get two samples when the bottom part drops off. Look up some of the standard electropolishing solutions and conditions. One is based on percholoric/butylcelsolve and the other on chromic acid/acetic acid -I think.
FEG note: Someone asked recently about FEG tips. One problem with these tips that anyone off the streets would have is getting material that is oriented. Normal W wires have a [011] texture. The desired orientation for emitters in order for them to have a low work function and high brightness is either [112] or [113] (I can't remember). Other than that, I think anyone with experience making FIM samples could probably duplicate their construction within a short time, even Schottky design.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Roberson, Michael (M) [mailto:MRoberson-at-dow.com] Sent: Friday, June 15, 2001 9:07 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Does anyone happen to know in what issue of microscopy Today the following information on sharpening W needles was found? (or how to make them) If so please send me a note at, MRoberson-at-dow,com
I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Michael Roberson (517)636-8656 SMX Analytical Sciences Midland, MI 48667
I installed the Lyson grayscale inks in our Epson 850N. The results are sometimes amazing, but always at least good.
Optimal results is greatly affected by paper type and quality, it seems more so than printing color. You will also need to fiddle with the colorspace gamma. Sorry I can't give more details, but I haven't had time to work with it too much since most people here are printing color. I think it has a great deal of promise and is worth checking out. The Lyson website has a number of tips and print comparisons for optimizing results.
As far as choosing printers, find people who have used any particular model. Epsons are slow to start printing. Out of the 4 epsons I work with (3 850N, 1 3000) and another 3 in a colleague's lab. at least 1, often 2, are misfeeding paper, engaging in randoms acts of stubborness, dropping off the network, etc. The 3000 is extremely touchy about how you load the paper, backing sheets, etc., and regularly soils itself with ink. Epson drivers have improved - sometimes, they even allow use to automatically scale to fit the page at print time. However, 2 of the 850N printers hasve never failed, always ready to go, so these problems probably reflect Epson QC issues..
Regards, Glen
"Hendrik O. Colijn" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Tom, } } I have seen a retrofit for Epson printers that looks quite interesting at } } http://www.piezography.com/ } } I haven't actually seen it in action but it looks like it should be good } for B/W prints. The advertising samples on the web site look very } nice. Basically they replace the color cartridges with different densities } of black ink. By allowing gray inks, they reduce the amount of dithering } required. The product consists of the replacement cartridges and the } revised printer driver software. } } Has anyone actually tried this approach? } } Cheers, } Henk Colijn } } At 12:00 AM 6/15/01 -0500, treese wrote: } } } We are going to replace our old Epson InkJet printer which produces nice } } B+W prints of micrographs, when it works. I haven't kept up with Epson vs } } HP for electron microscopists and wondered if a clear preference for one } } of these, or maybe another mfg or technology has emerged. We want B+W } } quality and reliability, and are willing to pay a bit more,and sacrifice } } speed and color quality...Thanks...Tom Reese } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://web.ceof.ohio-state.edu } Fools are pleased when they discover error. } The wise are pleased when they discover truth.
-- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax ************************************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
I have always used Epson and have never been disappointed with its best performance, but I do have an occasional problem with banding. I am on my third model, an Epson stylus 900, over the last 6 years. It is of the 1440 DPI Varity. Now they have 2880, although I am usually happy with 720DPI. I get the impression that HP's asset is reliability. HP has not focused on photo image quality as much as Epson, but both printers have surpassed the photo-realistic hurdle a few years ago, not to say that they are as good as film of' course. It would be interesting to buy both, do a side by side comparison and send the one that fails back. Let me know what you find.
On a side note I have been playing around with wide format prints. There is some good in printing larger hard copies and scanning at lower magnification to get a better representation of the surface, but where the limits are I don't know.
Don't write off speed too much. Speed usually only comes at the cost of dollars, not quality with these inkjets. I like USB as well, may be faster.
Ric
-----Original Message----- } From: treese [mailto:treese-at-marinebio.mbl.edu] Sent: Friday, June 15, 2001 1:00 AM To: Microscopy-at-sparc5.microscopy.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tom -
I've gotten quite passionate about marine invertebrate macrophotography as a retirement project (see the URL below if you like such things). I have a lot of prints on display in various local educational venues, so I've been very concerned with print longevity - at a reasonable cost, since I buy the equipment with my own $$$. I suggest that you look at http://www.luminous-landscape.com/1280.htm for a critical review of the Epson 1280. Epson's 6-color process is supposed to give 25-year color prints & 100-year B&W (see http://www.wilhelm-research.com/). I've just bought a closeout 1270, for $250; 1440 dpi rather than 2880, but what a deal!
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Thank you for all your replies to my question regarding oil red O. Here is my attempt to summarize those replies (and resulting discussions) starting with the earliest reply. Please forgive the haphazard manner of organization.
Jaclynn Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314.977.0257 fax: 314.977-0030 Jaclynn M. Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314.977.0257 fax: 314.977.0030
Ms. Allan-Wojtas,
In response to your request regarding oil red O lipid staining, here are the replies I received (along with some discussions as well). I have not had time to thoroughly digest everything myself, so please forgive the delay with which I replied and the lenghthy and unorganized manner of compiling the replies (listed in the order in which they were received).
Jaclynn M. Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314.977.0257 fax: 314.977.0030
---------------------------------------------------------------------------- --------------------------------------- } Has anyone used Oil Red O to stain lipids in tissues embedded in plastics } (Epon or Epon-Araldite)? If so, has this been done by staining en bloc or } by staining the sections. Sections would range from 1-4 microns in } thickness.
After the tissue is embedded, the solvents used (alcohol, propylene oxide) will have dissolved most or all of the lipids so there will not be much to stain. If you stain before embedding, dehydration will remove the lipid and the stain.
} We would also consider tissues embedded in glycol methacrylate.
Forget it if you are using alcohol (or acetone) for dehydration. I don't know of any dehydrating agents that will not remove lipids. Perhaps freeze drying would work, but you still need an embedding media that won't dissolve lipids. Even after osmium, some lipid is lost in dehydration.
} We'd like } to avoid frozen sections because we'd prefer the higher level of detail } possible with plastic.
So would a lot of us! This is why frozen sections are used for lipid staining.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu ********************************************** ---------------------------------------------------------------------------- -------------------------------- The lipids will be removed by the solvents used in processing. In tissues post fixed with osmium tetroxide BEFORE embedding in these plastics, the osmium will stain lipids black.
Gayle Callis MT,HT,HTL(ASCP) Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610
406 994-6367 404 994-4303 (FAX) ---------------------------------------------------------------------------- ----------------------------------- The problem with trying to do lipid staining in epon or plastic embedded tissue is that inorder to embed in epon or gma you usually have to process in solvents which will dissolve the lipids not to mention that the plastic itself is a solvent, at least GMA is somewhat.
Patsy Ruegg ---------------------------------------------------------------------------- ------------------------------------ I don't know anyone who has been able to "successfully" stain enbloc or sections from resins with oil red O, Sudan black or any of the other common lipid stains because the solvents used in the processing are all designed to remove fats and lipids. The only way I could get true accurate staining was with cryo sections. I have tried so many methods/ resin schedules/ types of resin and the same problems always occur, if there is enough lipid remaining to stain, it doesn't give a true representation of location and quantity. I'm sorry if this doesn't help, other than to perhaps save you some time and effort. As always, if you do happen to find someone who swears by their method, I would love you to email me a copy of it. Cheers, Kerrie
Kerrie Holmes Histology, Microscopy Research Research Division, Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett St. East Melbourne 8006 Phone: 9656 1244 / 1242 Fax: 9656 1411 Email: k.holmes-at-pmci.unimelb.edu.au ---------------------------------------------------------------------------- ----------------------------------- I do alot of samples in GMA and have tried oil red O on a couple of occasions with no luck. I would be very interested to know if anyone has done this and using what method. Good luck. Regards Liz
Elizabeth Cox Fisheries Biologist Queensland Department of Primary Industries Northern Fisheries Centre PO Box 5396 Cairns, Queensland, Australia 4870 Fax: 07 4035 1401 Ph: 07 4035 0158 ---------------------------------------------------------------------------- --------------------------------------- Has anyone used Oil Red O to stain lipids in tissues embedded in plastics (Epon or Epon-Araldite)? If so, has this been done by staining en bloc or by staining the sections. Sections would range from 1-4 microns in thickness.
A couple of people so far have commented on how hard this would be, and our experience agrees with theirs. HOWEVER Jaclynn also said:
We would also consider tissues embedded in glycol methacrylate. We'd like to avoid frozen sections because we'd prefer the higher level of detail possible with plastic.
In THAT case things look better! Would you settle for Sudan black, rather than Oil red staining of the lipid? If 'yes' then there is a method - which does indeed show even tiny droplets of lipid very clearly. This was worked out by the one-time king of GMA staining Peter Gerrits, and can be found in J Neurosci Methods, as follows:
Gerrits PO, Brekelmans-Bartels M, Mast L, 's-GrAavenmade EJ, Horobin RW and Holstege G. (1992).. Staining myelin and myelin-like degradation products in the spinal cords of chronic experimental allergic encephalomyelitis (Cr-EAE) rats using Sudan Black B staining of glycol methacrylate-embedded material. J. Neuroscience Methods. 45, 99-105
Bye - Richard Horobin
Institute of Biomedical & Life Sciences, University of Glasgow T direct 01796-474 480 --- E RichardWHorobin-at-aol.com "What should we expect? Everything." ---------------------------------------------------------------------------- ------------------------------------ Richard-
Whenever I've worked with those plastics, there has always been a clearing stage through acetone. Since acetone would remove all non-bound fat, Oil Red O would have nothing to go into.
When I've worked with these plastics, I also did a post-fixation in osmium tetroxide, which does a very good job of staining fats and lipid (it turns them black). Perhaps this would work for your purposes.
Joe
Joseph A. Saby, BA, HT(ASCP) Drug Safety Evaluation Pfizer Global Research and Development 2800 Plymouth Road Ann Arbor, MI 48105 Phone: (734)-622-3631 FAX: (734)-622-3866 E-mail: joseph.saby-at-pfizer.com ---------------------------------------------------------------------------- -------------------------------------- Certain stains for light microscopy are not usable if you are trying to stain for lipid and increase resolution of the sectioned tissue. One has to adjust one's thinking and look only at how lipid can be preserved AND processed into a plastic, which involves solvents such as alcohol during the dehydration and infiltration steps. If you are an experienced EM person, you have probably noticed that lipid has been seen as a green color in your semithin secions. If you look at Toluidine Blue stained semithin sections, lipid remains green, and the nuclei and cytoplasm are blue. The resolution of epoxy and the green color allows for subcellular identification of lipid.
HOWEVER, the preservation of lipid by osmium can be greatly enhanced by p-phenylenediamine (use carefully, it can cause contact dermatitis and asthma). Osmicate normal time, then start you dehyration procedure 25%, 50%,then put tissue in a 1.0% p-phenylenediamine in 70% ethanol for 15-25 minutes, then finish dehydratation procedure as normal, up to 100% ethanol, into ethanol/propylene oxide, etc....Semithin (1-4u)sections without any additional staining will show lipid by light microscopy. If needed, counterstain nuclei with methylene blue stain or Toluidine Blue.
Glycol methacry. would not work, because none of the lipid will remain, due to the alcohols used in processing. Only osmication, and especially post- osmication treatment of tissue with p-phenylenediamine perserves and stablizes lipid so it doesn't wash out during dehyration steps.
Good Luck!
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642 ---------------------------------------------------------------------------- ---------------------------------------------- Years ago I prepared a demonstration slide of testes for the Leydig cells that came from a conventional EM block, Glut/Cacod and Osmium. 1 micron section stained with Sudan black then Toluidine blue. Beautiful result but it was only a one-off, although something I'll try again if needed. Ian.
Dr. Ian Montgomery, West Medical Building, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855. Extn:6602. Fax: 0141 330 2923 e-mail: ian.montgomery-at-bio.gla.ac.uk ---------------------------------------------------------------------------- ---------------------------------- A question: is this really the case? Osmium binds across double-bonds of lipids, which is why it shows and preserves membranes, but it binds poorly or not all at to saturated lipids. So I wouldn't expect OsO4 to show fatty deposits if the fats are saturated. Oil Red O and Sudan Black, which more mix into the lipids and don't react with them, would show fat deposits that OsO4 doesn't, and are better fat stains.
Phil
} Agreed, there would be no need to do special fat stains if the tissue is } processed with osmium. In fact, this can be done for paraffin embedding as } well to show fat distribtion in some diseases with vastly better morphology } and localization than frozen sections. } } Tim Morken } CDC, Atlanta ---------------------------------------------------------------------------- --------------------------------------- Do you have any fixed, unprocessed tissues to work with?? If so, do frozens on them instead of your blocks. This is what all the ORO protocols that I have say to do, which makes sense when you consider that all processing will subject the tissues to lipid solvents, thus all or most of the lipids will have been removed and plastics are especially good lipid solvents.
Connie McManus ---------------------------------------------------------------------------- ---------------------------------------- The block was post fixed with osmium tetroxide/cacodylate then stained with Sudan black. It was only a wee trial just to see what happened. At the time I naively thought that the Sudan black must be binding to the osmium fixed fat. It was only a once of on a single slide. I stained another slide with Toluidine blue/Pyronine Y and it was just as lovely. Goodness knows what the answer was. At the time I used to try all sorts of staining combinations on semi-thin Glut/Osmium resin sections. Some were awful but others showed promise, unfortunately time didn't permit further investigation. The best staining for Glut/Osmium resin sections, in our hands was Toluidine Blue/Pyronine Y (Ito & Winchester 1963 J. Cell Biol) and a Polychrome technique (Pasyk, Bartok & Fabry 1989 Stain Technol 64 (3) 149. Ian.
X-Sender: uvsgc-at-trex2.oscs.montana.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32)
MicroListers,
This is a timely topic for me. We are getting a new chiller for our two EM's and I'm reconsidering what to use in it. For over 20 years I've run them with tap water and a dusting of dichlorophene fungicide on the surface of the water in the tank, which powder slowly dissolves into the water. Have had no problems at all with corrosion, plugged up lines (well, only ONCE, when I'd failed to add fungicide after water change), so I'm inclined to keep doing that, pending manufacturer's recommendations when I get the new chiller.
However, other voices have advised using 10% propylene glycol (pure stuff, NOT as in automobile anti-freeze) in de-ionized water.
Would like to revisit the pro's and con's of each method.
Thanks for your advice, in advance,
Gib Ahlstrand
} Haixin, } I've found that using distilled or de-ionised water in the chiller in } addition to the measures already suggested will also greatly reduce the } growth of microorganisms by denying them nutrients. } } Regards, } Eric lachowski
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
Just out of curiousity. . . how much X-radiation penetrates the rubber stopper?
Marie
Dear Marie, The short answer is, "Nearly all of it." The flux of x-rays depends on such parameters as the location of the stopper with respect to things in the column which can scatter the incident beam, produce brehmsstrahlung, etc. The easiest thing to do is to take a counter and make the measurement. For x-rays, the appropriate detector is an ionization chamber, such as a hand-held QT-Pi; however, a Geiger counter can also be used to give a relative measurement between the original set-up and that with the stopper. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Luminos http://www.lumijet.com/mono.htm also has quadtone inksets for many Epson printers. With any of the quadtone type ink systems, you will really need to dedicate the printer to printing only black and white. It is impractical to switch between color and monochrome ink sets as the printer needs to be flushed out before each switch.
I have seen some "fine art" photo samples printed with this ink system and the results are stunning.
George
George Laing National Graphic Supply v:(518) 438-8411 X3109 f:(518) 438-0940 email: scisales-at-ngscorp.com } } Tom, } } I have seen a retrofit for Epson printers that looks quite interesting at } } http://www.piezography.com/ } } I haven't actually seen it in action but it looks like it should be good } for B/W prints. The advertising samples on the web site look very } nice. Basically they replace the color cartridges with different } densities of black ink. By allowing gray inks, they reduce the amount of } dithering required. The product consists of the replacement cartridges and the } revised printer driver software. } } Has anyone actually tried this approach? } } Cheers, } Henk Colijn
I too have heard this tick from a company in long Island NY. I called, got sample prints, looked good, but I could tell better with my own images. It also might be cheaper since you can do continuous feed on some Epson products and buy ink in bulk.
-----Original Message----- } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] Sent: Friday, June 15, 2001 8:09 AM To: treese; Microscopy-at-sparc5.microscopy.com
Tom,
I have seen a retrofit for Epson printers that looks quite interesting at
http://www.piezography.com/
I haven't actually seen it in action but it looks like it should be good for B/W prints. The advertising samples on the web site look very nice. Basically they replace the color cartridges with different densities of black ink. By allowing gray inks, they reduce the amount of dithering required. The product consists of the replacement cartridges and the revised printer driver software.
Has anyone actually tried this approach?
Cheers, Henk Colijn
At 12:00 AM 6/15/01 -0500, treese wrote:
} We are going to replace our old Epson InkJet printer which produces nice } B+W prints of micrographs, when it works. I haven't kept up with Epson vs } HP for electron microscopists and wondered if a clear preference for one } of these, or maybe another mfg or technology has emerged. We want B+W } quality and reliability, and are willing to pay a bit more,and sacrifice } speed and color quality...Thanks...Tom Reese
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Fortunately, the EDS port is to the rear of the column and pointing upward.
Dear Earl, Fortunate for the user; however, an often-neglected aspect of radiation shielding is that sufficiently energetic x-rays will penetrate floors and ceilings as well as walls, so check with the folks upstairs if you have a TEM--especially an IVEM or HVEM. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I'd love some feed-back from someone who knows radiation because I've felt that applying TEM rules to SEMs is gross over-kill, especially with so many color monitors around. How often do you have your decelerating grid checked for proper operation? If it doesn't operate correctly, your exposure could be very dangerous, given the time that one sits in front of these things and their distance from your face.
If I'm way off base, I'd like to know and know why.
Dear Ken, I think I qualify as someone who knows radiation, so here goes. Yes, TEMs and SEMs are different, but the standards for stray radiation should be the same. It is a lot easier to meet the standards with a low-voltage, low-current machine, so the necessary shielding for a SEM would be less difficult than for a TEM, but there should still be less than the specified x-ray flux at the surface of the column. The Electron Microscopy Safety Handbook (2nd Ed., 1994), p 49 gives a standard set by the Radiation Control for Health and Safety Act of 1968 as 0.5 mR/hr at 5 cm from the surface of the column. I am not aware of any update of this standard, but if there is, the exposure allowed would surely be lower. A hand-held ionization chamber can be used to measure the radiation from both the microscope and the color monitors. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi Michael, I believe that the article in Microscopy Today that you are looking for is "Preparation And Use Of Needles and Micropipets For Handling Very Small Particles" by Anna Teetsov of McCrone Research Institute. It is a 4-pager that I am faxing to you. I would be pleased to fax copies to anyone else on the list that would like a copy. Best to all, Don Grimes, Microscopy Today
-----Original Message----- } From: Roberson, Michael (M) [mailto:MRoberson-at-dow.com] Sent: Friday, June 15, 2001 8:07 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Does anyone happen to know in what issue of microscopy Today the following information on sharpening W needles was found? (or how to make them) If so please send me a note at, MRoberson-at-dow,com
I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Michael Roberson (517)636-8656 SMX Analytical Sciences Midland, MI 48667
I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help.
Dear Haixin, We use dichlorophene to prevent the growth of micro-organisms, but you may also need to remove those which are already in your system. The appropriate cleaner will depend on what the components of the system are made of. If you can disassemble and clean out the hoses, lenses, and chiller separately, you would probably be able to get things cleaner, but it could be a big effort. Cu++ is toxic to algae, and will not usually damage metals, so you might want to circulate a solution of CuSO4, pH adjusted to ~8 through the lenses if the lines are constricted within the lenses. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Whew! I am the only one in our EM lab. About 90 - 100 Kidney/surgical/autopsy specimens a year. About 30 or so research specimens a year. Not overworked here, but sure wish I had some backup, as vacations or sick days are a nightmare. I come in all hours, too.
Marilyn Howton EM lab Pathology Department WVUniv. Hospitals
Does anyone have a current email address for Sycon Instruments (makers of film thickness monitors)? Their web site is still up, but with a 1998 date, and the group listed as maintaining the site is apparently defunct. No email addresses are listed on the Sycon web site.
Thanks.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
I recently got rid of 2 Zeiss EM 9s2 microscopes and am interested in finding if there is anyone out there who is interested in buying 4489 EM film for a Zeiss 9s2? I have aluminum cassettes, steel cassettes and ~10 boxes of film I wish to sell.
Please respond via my email address!
Cheers! Ken ------------ Ken Tiekotter, Adjunct Professor The University of Portland Dept. of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Fortunately, in my lab, the administration was upstairs so not held in high regard & the original question was for an SEM.
In addition, the rubber stopper solution was meant to be a temporary situation. Given that the maximum 25 - 30 KV X-rays need to penetrate several layers of aluminum foil as well as concrete floor I doubt that it poses a health risk. We had this situation at the last lab I worked & the Safety Department measured the X-ray output and gave it a clean bill of health. Not a good permanent situation but OK for the month or two.
Best Regards,
Earl
----- Original Message ----- } From: {"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, June 15, 2001 11:33 AM
Hello Microscopists,
We all work very hard at producing beautiful and relevant microscope images. Now is your chance to submit your finest images to the "Small World 2001" contest. This is the 26th year for the famous contest that results in a beautiful calendar showcasing the winners.
The closing date for submitting your images is June 29th, 2001. This can now be accomplished easily and quickly via the http://www.microscopyu.com/smallworld/ website.
There's still time to enter this famous optical photomicrography competition which is open to both professional and amateur microscopists; 35mm slide and/or digital entries of images taken with any kind of light microscope are accepted. Follow this link -
http://www.microscopyu.com/smallworld/
to learn more about the contest, see the fabulous prizes available, obtain your "Small World Screen Saver" of last years winners and view some of the stunning images that have won in the past.
Also take time to browse the MicroscopyU host site which has a wealth of exciting microscopy tutorials and resources. Follow the host link by typing this address into your browser -
http://www.microscopyu.com/
Good Luck and Best Regards,
Stan Schwartz Manager, BioScience Dept. Nikon Instruments Inc. 1300 Walt Whitman Rd. Melville, NY 11747 Phone: 631-547-8529 Fax: 631-547-4033 email: sschwartz-at-nikon.net www.nikonusa.com Check out Nikon's Educational Website www.microscopyU.com
A nice plug for Tina's web pages from the NYTimes.
NYTimes 06-14-01
What You Can't See Microscopically
By SHELLY FREIERMAN
If you like to ponder questions on the scale of how many Web sites can fit on the head of a pin, there is a fine collection of electron microscope images to examine at MicroAngela, an online collection (www.pbrc.hawaii.edu/bemf/microangela). The Web site is maintained by Tina M. Carvalho, the supervisor at the Biological Electron Microscope Facility at the University of Hawaii in Manoa.
She has taken the images of tiny bugs, bacteria, mold, parasites and grains of sand, and colored them.
There is a description for each of the objects, which were collected from captured insects; from samples of ocean life, like tubeworms and coral; and even from mold growing on some Romano cheese in Ms. Carvalho's refrigerator.
As indicated earlier (5-13-2001), I am soliciting suggestions for symposia for the year 2002. So far we had very good response and I am confident that the program for 2002 will be excellent. However, before the program will be finalized, I would like to once more ask for your idea for a great topic in any of the areas of
"Physical Sciences" "Biological Sciences" "Advances in Instrumentation and Techniques"
The deadline for suggestions is June 30th 2001. Please contact me with your suggestions before June 30th under the following address:
mm2002-at-ornl.gov
Please forward this request also to your colleagues. Thank you for your support.
With best regards,
Edgar Voelkl Program Chair M&M 2002 --
___________________________
Dr. Edgar Voelkl Program Chair M&M 2002
Oak Ridge National Laboratory P.O. Box 2008 Bldg 4515 Oak Ridge, TN 37830-6064
I've had Epson and Lexmark printers in the past, but can't say enough about the HP Photosmart 1215 I've recently purchased. I've always had paper feed problems with the Epson and Lexmark printers I've used, and noticed that the HP I used on my home computer didn't have those problems. I decided to try an HP printer for my business computer and chose this one. For both color and grayscale images, I find it to be the closest to photographic quality that I've tried (I used to do professional photography in the arts field and did all of my own developing and printing).
The only caveat is that grayscale images should be printed in color mode - black only prints in 600 x 600 resolution while color prints in 2400 x 1200.
This printer includes a separate 4 x 6 inch paper feed tray that is excellent for small prints.
On Friday, June 15, 2001 12:00 AM, treese [SMTP:treese-at-marinebio.mbl.edu] wrote: } ------------------------------------------------------------------------ } } } We are going to replace our old Epson InkJet printer which produces } nice B+W prints of micrographs, when it works. I haven't kept up } with Epson vs HP for electron microscopists and wondered if a clear } preference for one of these, or maybe another mfg or technology has } emerged. We want B+W quality and reliability, and are willing to pay } a bit more,and sacrifice speed and color quality...Thanks...Tom Reese }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} } I've had Epson and Lexmark printers in the past, but can't say } enough about the HP Photosmart 1215 I've recently purchased. I've } always had paper feed problems with the Epson and Lexmark printers } I've used, and noticed that the HP I used on my home computer didn't } have those problems. I decided to try an HP printer for my business } computer and chose this one.
Yeah, maybe, but boy, don't extend this to all HP printers.
I used to have at home an HP 400 inkjet, the cheapie, which kept on giving paper-feed problems until I donated it to the repair shop rather than fix it a second time, now, at work, I have a Laserjet 6L that often feeds thru up to 10 sheets at a time!
I can't load up the input stack, have to insert each sheet to be printed, one at a time.
It seems to me that the HP problem is that, in order to keep the footprint small, the paper has to turn almost 180 degrees in a tight circle.
The 6L has at the moment ANOTHER paper jam that I feel too angry to fix, it's been jammed now for two days but I'd rather print to the network printer and walk down the hall to retreive the printout than pull it apart AGAIN.
If I had an outside window I'd be tempted to throw the *!-at-#$% printer through it....................................
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Thank you all very much for the reply of my question. I think I figoured out how to deal the problem.
Haixn Xu
Biological Sciences/UMBC Baltimore Maryland
On Fri, 15 Jun 2001, Beauregard wrote:
} Hi, } } Chloramine-T can be used to kill or prevent the bacteria but it will cause } corrosion problems with some instruments over time. This is especially } true of } stainless steel fittings where pit corrosion can become a very serious } problem and it was for us. The manufacturer recommended using pure } distilled water in our TEM. We didn't do what the service people said } because we never had the problem with our other instruments in our wing of } the building with C-T. We fixed our problem by using some manufacturer } supplied brass fittings. We also switched to de-ionized water. It worked } fine. } } Anyway, you could kill them with C-T, flush the system completely and then } switch to pure distilled or de-ionized water. Check with your serviceman } for the recommended material to use. } } After flushing, you can monitor the chloride level and the available } chlorine to be sure you have removed the chloramine-T. You can use a } swimming poll test kit for available chlorine and 0.1N silver nitrate } solution for testing for chloride ions. } } After flushing and switching to pure distilled water our corrosion problem } has not surfaced in over 8 years. We have about 10 instruments on our } recirculator system. } } One further note. You can get a blue-green to green color in the water and } inside the water lines. This can be copper ions or a green carbonate, } like copper carbonate. Check any transparent PE or PP lines to see if they } have a } coating inside them of green copper carbonate. I did this on our Edwards } 306 evaporator because it had transparent water lines on the DP. This } color was thought to be algae but it was not algae alone. You can scrape } off some of the coating and under a microscope apply hydrochloric acid. } Look for any bubble generation to confirm the possible presence of copper } carbonate. Of course, the bubbles could be another material like calcium } carbonate but it would not ordinarily be greenish. Remove one of the lines } if you can, add dilute HCl, wait for the line to 'clear' and run the acidic } liquid into a test tube. If it's blue, you probably have copper ions and } most likely copper carbonate deposits. We did. The copper can be } confirmed by atomic absorption if necessary. } } Hope this helps. } } Paul Beauregard } Senior Research Associate } PPG Industries } Monroeville, PA 15601 } } } } At 05:22 PM 6/14/01 -0400, Haixin Xu wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Hi lister, } } } } I have some problems with our EM chillers. The water in EM cooling } } circulation is bad contaminated by algae, bacteria, maybe more } } microorganisms. Does somebody here know some water cleaner for chillers of } } EM? Appreciate for the help. } } } } Haixin Xu } } } } Biological Sciences } } University of Maryland, Baltimore County } } Baltimore, Maryland } } } } } } } } } }
Hi Don, I would appreciate a copy, if you wouldn't mind. I have a set-up of my own, but it is quite the "homemade" variety.
Using this set-up, I have also produce fine blades by using flattened W wire rather than the rods. Carefully dipping them in a particular way produces a blade on one side and leaves the other side as a support for strength. Works well when slicing nematodes. John Shields EM Lab Univ. of GA, Athens
On 15 Jun 2001, at 15:23, Don Grimes wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of
} Hi Michael, } I believe that the article in Microscopy Today that you are looking } for is "Preparation And Use Of Needles and Micropipets For Handling } Very Small Particles" by Anna Teetsov of McCrone Research Institute. } It is a 4-pager that I am faxing to you. I would be pleased to fax } copies to anyone else on the list that would like a copy. Best to all, } Don Grimes, Microscopy Today
Many thanks. I now have the contact information for Sycon Instruments.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
} -----Original Message----- } From: "wft03-at-health.state.ny.us"-at-sparc5.microscopy.com } [mailto:"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com] } Sent: Friday, June 15, 2001 3:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: water cleaner for EM chiller } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } } } } I have some problems with our EM chillers. The water in EM cooling } circulation is bad contaminated by algae, bacteria, maybe more } microorganisms. Does somebody here know some water cleaner } for chillers of } EM? Appreciate for the help. } } Dear Haixin, } We use dichlorophene to prevent the growth of } micro-organisms, but you } may also need to remove those which are already in your system. The } appropriate cleaner will depend on what the components of the } system are } made of. If you can disassemble and clean out the hoses, lenses, and } chiller separately, you would probably be able to get things } cleaner, but } it could be a big effort. Cu++ is toxic to algae, and will } not usually } damage metals, so you might want to circulate a solution of CuSO4, pH } adjusted to ~8 through the lenses if the lines are } constricted within the } lenses. Good luck. } Yours, } } Bill Tivol } Wadsworth Center } Albany NY } (518) 473-7399 WFT02-at-health.state.ny.us } }
We recently acquired an older Link EDS spectrometer and would like to use it on an old ESEM. But we will need the pulse processor and whatever else to run the detector. I have heard of people mixing brands and we have some other EDS systems, but our old Link is presently installed on another scope. Does anyone have an old Link giveaway or have any thoughts on compatibility among the EDS systems? Thanks for any thoughts or assistance. Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
This is an update on a possible way of quantifying the orientation of actin filaments within endothelial cells in culture.
I would like any comments on whether this seems reasonable or if there are potential problems.
The fluorenscent labelled cells are captured and saved as grayscale images. A highpass filter is used to enhance the filaments. The filaments are thresholded and skeletonized. then the branch points are selected and deleted. This creates many short linear segments whos "moment angle" or orientation can be measured. There are about a 500 - 1000 measurements per cell. The histogram of the angles have peaks cooresponding to the dominant directions of filaments. I am hoping that control and experimental groups of cells will have appropriatly different histograms.
I have a DURST S-45 enlarger that has not been used for a long time. The mirror is heavily dusted. I want to clean it or want it cleaned. But I am afraid to damage it if clean it not properly and I do not know where and who can do the job for me. I am located in Baltimore. I would very much appreciate your help.
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, MD 21250
We are looking for a used Ultracut T that is in good/excellent condition. Please contact Kathleen Greer or Lauren Simmerman if you know of one, -at- Univ. of Nebraska Med. Cen. 402-559-7729 or KathleenGreer-at-nhsnet.org
Paper jams mostly related to the paper quality (if printer itself is OK). You may ask manufacturer which particular type of paper is better for printer or just try different brands. For laser printers it should be mentioned on the paper that this is for laser. Paper for laser and ink -jets are opposite: glossy and "full-body" for laser and more porous (to adsorb inks) for ink-jets. It also important to set correct paper-thickness on the printer. They usually has thick-medium-thin settings. It's mechanical switch, which presents on most models.
We do have some LaserJet B&W printer running for 5 years without any serious problem with paper (heavy loaded, it serves as a network printer for whole Department). We just bought a new Tektronix Color-laser last year and it had jam problems near every day. We did change the paper and it works very good now. Same - for ink-jets.
Sergey
At 05:49 PM 6/18/01 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I have a DURST S-45 enlarger that has not been used for a long time. The mirror is heavily dusted. I want to clean it or want it cleaned. But I am afraid to damage it if clean it not properly and I do not know where and who can do the job for me. I am located in Baltimore. I would very much appreciate your help.
Dear Haixin, I would first remove the mirror (if possible), then gently blow as much of the dust off it as possible. The concern, of course, is that some of the dust may consist of mineral particles capable of scratching the mirror. These should be somewhat denser and more easily removed in an air stream than some of the other dust components. I would then rinse the mirror with a stream of water and/or dunk it into a warm detergent solution, then rinse carefully in distilled water followed by alcohol and/or acetone. In none of these steps would I wipe the mirror even with a very soft cloth. After this, I'd check to see how much dust is left. If none, I'd clean the mirror with optic-wipe cloth, if little, I'd re-rinse and do so until as much of the dust was removed as possible. I'd then clean a small area with optic wipe cloth and inspect for scratches, if none, then I'd clean the rest. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Reply to Haixin Xu: Cleaning is done the same as it would be for a front surfaced telescope mirror as follows: 1) Gently let pure water (distilled or deionized) flow over the mirror. 2) dip several "Q-tip" swabs in a 1:100 diluted solution of Triton X 100. 3) allowing only the weight of the wet"Q-tips" to be the downward force gently pass them over the mirror and rinse with more pure water and allow to dry dust-free. If only traces of dust settle on the mirror you can blow them off gently with canned gas or a rubber bulb duster.
Below is the result of your feedback form. It was submitted by (johnsond-at-ns.neiu.k12.pa.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 18, 2001 at 13:44:52 ---------------------------------------------------------------------------
Email: johnsond-at-ns.neiu.k12.pa.us Name: Debbie Johnson
Organization: Valley View School District
Education: 6-8th Grade Middle School
Location: Archbald. PA USA
Question: What power of magnification would we need to be able to see bacteria cells? They are not very clear using the standard classroom scopes.(400x) Are there microscopes available for classroom use that have more magnification?
I was wondering if anyone out there had any information on a morphometry or image analysis user group that I could get in contact with. Also does anyone have an interest in a confocal microscope MRC 600 that we are going to decommission. I would appreciate any information anyone could offer on any of the above subjects.
Hi Listers, Is there anyone out there who can help with this request? Please reply to plant-tc-at-tc.umn.edu. Thanks, Priscilla *****************************************************************************
----- Original Message ----- } From: Abhay Shendye To: plant-tc-at-tc.umn.edu Sent: Sunday, June 17, 2001 12:05 PM
Hi Mike,
We have had this done the other way (Link have run another maker's detector). They had to change the preamp onwards and ensure that they got the pinout and bias voltage to the crystal correct (obviously). We have also had upgrades and they have always changed the preamp with the pulse processor. What about contacting your local Oxford Instruments rep. to see what they have from recent part exchanges or upgrades?
Good luck, Ron
} We recently acquired an older Link EDS spectrometer and would like to use it } on an old ESEM. But we will need the pulse processor and whatever else to } run the detector. I have heard of people mixing brands and we have some } other EDS systems, but our old Link is presently installed on another scope. } Does anyone have an old Link giveaway or have any thoughts on compatibility } among the EDS systems? } Thanks for any thoughts or assistance. } Mike } ******************************************************************** } Michael L. Boucher Sr. mboucher-at-tc.umn.edu } Lab Manager Rm 55 Office Ph 612-624-6590 } I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 } University of MN Fax 612-625-5368 } 12 Shepherd Labs } 100 Union Street S.E. } Minneapolis, MN 55455 http://resolution.umn.edu } ******************************************************************** } } } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
For EM guys it's not big deal to restore even scratched mirror. Only limitation is the size of your vacuum evaporator's vacuum chamber.
1. remove old coating (very likely, it was aluminum, if so, use 1N NaON); 2 rinse in d-H2O and dry in clean place; 3. evaporate a new layer of Al (or Cr) using your vacuum evaporator.
The trick: never touch working surface before and after.
Sergey
P.S. Organic solvents may leave the traces even if they are very pure (alcohol may have the trace of CuSO4 used for dehydratation or something else). Last step in cleaning should be dd-H2O and drying in very clean place. Who wear glasses half of life knows, that any organic solvents leave the traces except detergent and then d-H2O. At least, it's my experience.
At 05:24 PM 6/18/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers, I want to take the microstructure of 0.75mm wire, which is used by dentist. Can anyone can guide for the sample preparation of wire for tem.
Arti Harle Regional Sophisticated Instrumentation Center Indian Institute of Technolgy-Bombay Powai, Mumbai- 400076. India. Phone No. 91-22-5767691 Extn. 7694 email: aartih-at-usa.net
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If we assume your bacteria are 0.000001m long then if you magnify them x400 they will appear 0.0004m long ie 0.4mm.
So if you would like them to appear, say 400mm long, you would magnify by x40,000.
There are special microscopes called electron microscopes which can give you this magnification. We look at bacteria at x1,000 - x94,000. Your local university will have some - why not check their web site?
Dave
On Tue, 19 Jun 2001 01:55:56 -0500 johnsond-at-ns.neiu.k12.pa.us wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form. It was submitted by } (johnsond-at-ns.neiu.k12.pa.us) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June } 18, 2001 at 13:44:52 } --------------------------------------------------------------------------- } } Email: johnsond-at-ns.neiu.k12.pa.us } Name: Debbie Johnson } } Organization: Valley View School District } } Education: 6-8th Grade Middle School } } Location: Archbald. PA USA } } Question: What power of magnification would we need to be able to see } bacteria cells? They are not very clear using the standard classroom } scopes.(400x) Are there microscopes available for classroom use that } have more magnification? } } --------------------------------------------------------------------------- }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
To answer the question of compatability. The "older Link EDX" is probably an optical reset FET detector if it is older than 1988. If it is a transistor reset FET ( PentaFet) then it would have an attribute number on the Dewar label. Therefore when you are looking for a compatable analyzer you will need to know which FET is in the detector. The bigger problem is that Link used a software based Restore signal to reset the FET therefore only a Link analyzer system will operate the detector. The only present EDX company using a similar software based Restore signal is Oxford ( Which bought Link Analytical back in the early 1990's ) The pre-amp on the Link detector could be changed so that the Restore signal is generated by the pre-amp. This would allow almost any analyzer system to operate the detector but some compatability issues would still remain.( Inhibit and Gain ) I hope this answers your question. Kind Regards, Jim Nicolino X-Ray Optics / AAT 1816 St. Johns Bluff Road Suite 305 Jacksonville, FL 32246 PH 904 646-3069 FAX 904 646-3131 E-mail JNicolino-at-xraydetectors.com
"Michael L. Boucher" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We recently acquired an older Link EDS spectrometer and would like to use it } on an old ESEM. But we will need the pulse processor and whatever else to } run the detector. I have heard of people mixing brands and we have some } other EDS systems, but our old Link is presently installed on another scope. } Does anyone have an old Link giveaway or have any thoughts on compatibility } among the EDS systems? } Thanks for any thoughts or assistance. } Mike } ******************************************************************** } Michael L. Boucher Sr. mboucher-at-tc.umn.edu } Lab Manager Rm 55 Office Ph 612-624-6590 } I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 } University of MN Fax 612-625-5368 } 12 Shepherd Labs } 100 Union Street S.E. } Minneapolis, MN 55455 http://resolution.umn.edu } ********************************************************************
Did you try Hexane to clean-up EM parts? Hexane, HPLC grade is very nice for that. As a matter of fact, many organic solvents even very pure has some additives, which is not listed. It includes the traces of compounds, which were used during purification process. Very usual contamination - sort of mineral oil, which comes when you store your stuff in the plastic bottle (or it was stored somewhere before it comes to you in glass bottle) - manufacturers use it in press-forms when they make plastic bottles. Any plastic (even teflon) has some "plastifiers" to make plastic more flexible. Those compounds does not chemically bonded to the plastic matrix, so they erode slowly into stored solution. I had a deal with very precise mirrors from analytical UV centrifuge. Those mirrors has precision comparable with best telescope mirrors. I find, that many organic solvents leave traces even if it never happen before on other surface. The mirror surface, looks like so perfect and reflect easy very tiny imperfection. Of coarse, I am talking about mirrors with reflecting coating up. As for fingerprints - any organic solvent should work, I agree with you. Speaking honestly, in the practical life my precautions may be not necessary. I was talking in general, that scratches on the mirror is not a problem for serious electron microscopist.
Thanks for your reply and have a great day.
Sergey.
At 10:20 AM 6/19/01 -0400, you wrote:
} P.S. Organic solvents may leave the traces even if they are very pure } (alcohol may have the trace of CuSO4 used for dehydratation or something } else). Last step in cleaning should be dd-H2O and drying in very clean } place. Who wear glasses half of life knows, that any organic solvents } leave the traces except detergent and then d-H2O. At least, it's my } experience. } } Dear Sergey, } d-H2O should leave no residue, but I've seen problems using it as the } final rinse--probably because it takes a long time to dry, and our air may } not be that clean. We use acetone as the final rinse on all the metal } parts of the microscope that go in the vacuum, because it's easier to pump } down if there is no residual H2O. We also use 95% EtOH to clean the glass } plates in the Perkin Elmer microdensitometer--it is the best solvent for } fingerprints--and we do not rinse with H2O afterward. I haven't seen any } residue problems from either of these solvents. } Yours, } Bill
Hello fellow microscopists, What are the latest advances in confocal microscopy systems? Any with Dapi or Hoechst detection? What are the pros and cons of PMT vs Camera acquisitions? Aperture, Fixed slits or spinning discs? We would appreciate any input.
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Debbie -
Yes, there are microscopes that will provide an excellent 1000x image. BUT - they are quite a bit more sophisticated (and expensive) than the scope that you are using, and they require careful setup to produce a good image. It's possible that there is a microscopist who reads our ListServer who is near Archibald who would be willing to visit your class to do a demonstration. If you are interested, please write again.
You can find a lot of information about bacteria at the American Society for Microbiology's educational website, http://www.asmusa.org/edusrc/
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones).
The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
I need professional help................. (in many ways).
Thank you.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
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I am working on a bacterial protein which are very unstable polymerisation and depolymerization is within very short time. I want to do the Atomic Force microscopy for these proteins What is sample preparation for AFM And how can I stabilise the protein.
Regards Arti Harle
Ms. Arti Harle Regional Sophisticated Instrumentation Center Indian Institute of Technology-Bombay (IIT-B) Powai, Mumbai - 400076. India Phone : 91-22-5767691 Extn 7694 91-22-5720109/ 5721039 (Resi) E-mail : aartih-at-usa.net
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Hi, all,
This question has come up for us at a good time, with this ongoing discussion of printers.
We recently purchased an Epson 880 ink jet printer for our EM/LM lab, and are very happy with it. The black and white prints look excellent. We have tried a number of papers, and find that the Epson "Photo Quality Glossy Paper" and "Photo Paper" give the best results for our purposes.
We are wondering how to go about making our prints more permanent from these printers. If the prints have just been printed and aren't dry yet, or if they are dry and are in contact with any moisture, they smudge quite easily. Is there any coating - lamination or spray, or something else which we can use to protect the surface?
In connection with this, we have also found, especially when we have printed b/w prints, that if we leave them in a room with full spectrum fluorescent lights for a few days, that they begin to change colour, and start to take on a reddish tinge. The instructions that came with the paper say to not leave prints exposed to sunlight - but we're talking about room lights, here! Has anyone else found this happens to them, and is there any way of preventing/minimizing this effect without storing prints in a light tight bag????
Any help or suggestions would be much appreciated. As always, thanks for your help in advance.
Regards,
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Impact Analytical has an opening for a microscopist. Interested candidates are invited to reply.
Job Description for Microscopist IMPACT Analytical seeks an exceptional technical professional to become a member of our Microscopy group. The candidate will be expected to provide outstanding service to our customers on projects in the areas of microtomy, SEM, TEM, and AFM. The candidate will be responsible for the development of approaches to solve customer problems, as well as for projects from the time that samples are received until the final report is written. This will include communication with the customer with regards to clarifying project goals and methods, conducting the analysis, updating the status, generating the report, and following-up on customer requests. Experience in the microscopy of plastics is a plus. The ideal candidate will have at a minimum, a B.S. degree and 2 to 5 years of relevant experience. The candidate should have a sense of urgency, strong organizational skills, be detail oriented, and have very good laboratory technique. The candidate should also have excellent oral, verbal, and interpersonal skills, as well as familiarity with Microsoft based software. Please respond by e-mail to Ms. Cherie Hutter {mailto:hutter-at-mmi.org} or by mail to Ms. Cherie Hutter, Impact Analytical, Job Number 06-01, 1910 West Saint Andrews Road, Midland, MI 48640 to arrive no later than 8 July, 2001.
Kevin Battjes Impact Analytical Michigan Molecular Institute 1910 W. St Andrews Road Midland MI 48640
We've had the same problem on our large format printer. I don't know what the answer is either. I haven't gotten nearly as far as you have in terms of troubleshooting, but my next step would be to call the manufacturer (Epson, in our case). Did your manufacturer have any insight?
Angela V. Klaus
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Tue, 19 Jun 2001, John J. Bozzola wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying (rather unsuccessfully) to print grayscale images on a } large format color printer. The printer does a superb job with color } images but generates truly unpredictable, bizarre renderings with } grayscale (SEM, TEM) images (greenish/brownish/pinkish tones). } } The printer has the option to print using black ink only but, } obviously, no color can be used elsewhere. I have made just about } every color management adjustment imaginable but have come to the } conclusion that it is not possible to print accurate colors and } grayscale simultaneously. Is that so? } } I need professional help................. (in many ways). } } Thank you. } } JB } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ############################################################## } }
I too have noticed that the cyan ink that Epson uses is not very stable even under fluorescent lights. However, enclosing a print in a polyethylene sheet protector seems to improve the stability quite a bit.
Cheers, Henk
At 07:47 AM 6/20/01 -0400, Paula Allan-Wojtas wrote:
} Hi, all, } } This question has come up for us at a good time, with this ongoing } discussion of printers. } } We recently purchased an Epson 880 ink jet printer for our EM/LM lab, and } are very happy with it. The black and white prints look excellent. We have } tried a number of papers, and find that the Epson "Photo Quality Glossy } Paper" and "Photo Paper" give the best results for our purposes. } } We are wondering how to go about making our prints more permanent from } these printers. If the prints have just been printed and aren't dry yet, } or if they are dry and are in contact with any moisture, they smudge quite } easily. Is there any coating - lamination or spray, or something else } which we can use to protect the surface? } } In connection with this, we have also found, especially when we have } printed b/w prints, that if we leave them in a room with full spectrum } fluorescent lights for a few days, that they begin to change colour, and } start to take on a reddish tinge. The instructions that came with the } paper say to not leave prints exposed to sunlight - but we're talking } about room lights, here! Has anyone else found this happens to them, and } is there any way of preventing/minimizing this effect without storing } prints in a light tight bag???? } } Any help or suggestions would be much appreciated. As always, thanks for } your help in advance. } } Regards, } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 10 - October 19, 2001
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2150 (Includes room and board, text, handouts, supplies)
Application Deadline: July 25, 2001
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual
I am trying to repair our Sorvall critical point dryer. I would like to replace the needle valve assemblies as they are starting to leak. Does anyone know if parts are available for this system? If not, we will have to get it replaced.
At 07:37 PM 6/19/01 -0500, John J. Bozzola wrote: } I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones). } The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
Which printer, which operating system, and which program is doing the printing? When you say "color management" what exactly do you mean that you tried?
Offhand, this sounds more like a function of the printer driver and the program driving it. The program you're using to print may only be capable of sending down grey or color at a time. When it sends color, the grey gets treated (and color-dithered) as color. In pure grey mode, no color dithering is introduced.
A more sophisticated printing program, such as a desktop publishing program like Quark, may be capable of driving the printer in a smarter fashion.
Hey, I hear you, been there and done that! (HP - oi!). Finally, in desperation I took a sheet of fine emery or sanding cloth to the rubber rollers that move the paper through the printer - it worked! Try it, be patient and do every roller you can get to. I've had to do this more than once, but not often.I think the rollers pick up a "shine" from paper coatings. Let me know what happens.
Chin up, man!
Gib
Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} } } } I've had Epson and Lexmark printers in the past, but can't say } } enough about the HP Photosmart 1215 I've recently purchased. I've } } always had paper feed problems with the Epson and Lexmark printers } } I've used, and noticed that the HP I used on my home computer didn't } } have those problems. I decided to try an HP printer for my business } } computer and chose this one. } } } Yeah, maybe, but boy, don't extend this to all HP printers. } } I used to have at home an HP 400 inkjet, the cheapie, which kept on } giving paper-feed problems until I donated it to the repair shop } rather than fix it a second time, now, at work, I have a Laserjet 6L } that often feeds thru up to 10 sheets at a time! } } I can't load up the input stack, have to insert each sheet to be } printed, one at a time. } } It seems to me that the HP problem is that, in order to keep the } footprint small, the paper has to turn almost 180 degrees in a tight } circle. } } The 6L has at the moment ANOTHER paper jam that I feel too } angry to fix, it's been jammed now for two days but I'd rather print } to the network printer and walk down the hall to retreive the } printout than pull it apart AGAIN. } } If I had an outside window I'd be tempted to throw the *!-at-#$% printer } through it.................................... } } } cheers } } rtch } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
} I am trying (rather unsuccessfully) to print grayscale images on a } large format color printer. The printer does a superb job with color } images but generates truly unpredictable, bizarre renderings with } grayscale (SEM, TEM) images (greenish/brownish/pinkish tones). I think this is why color printing is CMYK and not just CMY. We typically try not to allow the printer to print greyscale images with the color setting on because of the off-tones. I think there really is no way to achieve perfect grey with the color cartridges. I wonder if it would be possible to trick the computer into printing only the greyscale images with the black cartridge and everything else in color. The other alternative would be to do a "two-pass" print with one print through on the grey images, and then reprint again with color. This would be VERY difficult on a poster printer, but a little easier with one of those smaller feedthrough inkjet printers. } } The printer has the option to print using black ink only but, } obviously, no color can be used elsewhere. I have made just about } every color management adjustment imaginable but have come to the } conclusion that it is not possible to print accurate colors and } grayscale simultaneously. Is that so? My only other suggestion is to continue with those color management solutions--do you have all the color profiles configured from monitor to printer? You've adjusted the file to CMYK gamut from RGB before printing? The only other suggestion I can make(short of having the full colorimeter and spectrophotometer calibration) is to make sure that the CMYK color settings on the individual greyscale images are all manually adjusted to absolute neutral.
I am a long time Tem person starting a new job doing both TEM and LSCM. Its very exciting learning LSCM but I need some suggestions for ordering probes. Our lab has a Kr/Ar laser and will be doing neuropathology research, mostly with fixed human brain samples, but also tissue culture cells both live and fixed. A vibratome is in our future. I was thinking of ordering alexa488nm-phalloiden, alexa568- streptavidin, and syto60 from molecular probes to start getting used to the scope and the lasersharp bio-rad software. The immunohistotech in the lab uses an ABC dtection system with Dab. The first probe should contrast the cytoskeleton well(stain F-actin), the second probe will label any primary Ab, and the third will bind to DNA and contrast the nucleus. Will using these three probes together be a good starting point to becoming adept at labeling and producing high quality images on the LSCM including optical sectioning and 3D (and 4D sometime in the future)? I welcome suggestions Thank you
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John,
we used to have the same problem - the B&W images would come out pinkish. We contacted HP Software Support about a year ago, and they pointed out that there isn't any way to isolate a specific image and make it grayscale, leaving the others in color. They suggested making the image a gray scale before importing into the document. So far, when staff have copied the image and pasted it right into the poster (a PowerPoint slide - enlarged to the size of the poster or half the size of the poster) - it seems to work. However, there were some special images that a few people had that didn't work. Good luck,
Alice.
Alice Dohnalkova Dep. of Environmental Microbiology Battelle, Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
-----Original Message----- } From: John J. Bozzola [SMTP:bozzola-at-siu.edu] Sent: Tuesday, June 19, 2001 5:37 PM To: Microscopy-at-sparc5.microscopy.com
I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones).
The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
I need professional help................. (in many ways).
Thank you.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
-----Original Message----- } From: John J. Bozzola [SMTP:bozzola-at-siu.edu] Sent: Tuesday, June 19, 2001 5:37 PM To: Microscopy-at-sparc5.microscopy.com
I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones).
The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
I need professional help................. (in many ways).
Thank you.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
Luminos http://www.lumijet.com has a spray called "Imagesheild" to protect prints. $18/ can. Quoting their website: "LUMIJET IMAGESHIELD will become your favorite print protector. It’s a convenient ozone-friendly low-odor aerosol spray that significantly improves wet-fastness. In addition to protecting delicate inkjet images from moisture, it also shields images from fingerprints and harmful UV rays. This unique product will not only help extend useful print life, it is totally transparent; preserving the original print surface characteristics. From dead matte to glossy, or from canvas to silver foil, you’ll never know it’s there."
} We are wondering how to go about making our prints more permanent } from these printers. If the prints have just been printed and } aren't dry yet, or if they are dry and are in contact with any } moisture, they smudge quite easily. Is there any coating - } lamination or spray, or something else which we can use to } protect the surface?
There were recently (last 6-8 months) some issues with color shift on some Epson papers. Epson determined the shift was from high concentrations of ozone, and not due to exposure to light. Epson recommends Heavyweight Matte or Epson Photo paper for maximum lightfastness.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com } } In connection with this, we have also found, especially when we } have printed b/w prints, that if we leave them in a room with } full spectrum fluorescent lights for a few days, that they begin } to change colour, and start to take on a reddish tinge. The } instructions that came with the paper say to not leave prints } exposed to sunlight - but we're talking about room lights, here! } Has anyone else found this happens to them, and is there any way } of preventing/minimizing this effect without storing prints in a } light tight bag???? }
Epson had a magenta ink fade problem. I have "heard" that it has been improved. Anecdotal reading suggests the paper chemistry vs. ink can have a bearing on colorfastness. Prints I have made in the last year or so on Eposn "photo quality inkjet paper" and Kodak "Picture paper" have held up reasonably well. I have also used Krylon clear acrylic spray to "fix" some prints. ...Not bad for moisture, insufficient data about help with fade. Use many light coats. If you really wet the surface, the paper can become translucent.
WoodyWhite McDermott Technology, Inc
{SNIP} } } In connection with this, we have also found, especially when we } } have printed b/w prints, that if we leave them in a room with } } full spectrum fluorescent lights for a few days, that they begin } } to change colour, and start to take on a reddish tinge. The } } instructions that came with the paper say to not leave prints } } exposed to sunlight - but we're talking about room lights, here! } } Has anyone else found this happens to them, and is there any way } } of preventing/minimizing this effect without storing prints in a } } light tight bag???? } } } }
Below is the result of your feedback form. It was submitted by (aiace_99-at-usa.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, June 20, 2001 at 10:45:27 ---------------------------------------------------------------------------
Email: aiace_99-at-usa.net Name: Massimo
Organization: None
Education: Graduate College
Location: Italy
Question: I am writing to enquire about a technical problem. I'd like to see with my microscope some cells coming from a dissected tissue, for instance a vegetable material. So, if I have such small tips into a drop of water and I want to fix, stain and mount in Balsam, how can I perform this operation without to loose that material from the slide? Could you give me some information or a reference where I could find the correct protocol? Thank you Massimo.
I am a long time Tem person starting a new job doing both TEM and LSCM. Its very exciting learning LSCM but I need some suggestions for ordering probes. Our lab has a Kr/Ar laser and will be doing neuropathology research, mostly with fixed human brain samples, but also tissue culture cells both live and fixed. A vibratome is in our future. I was thinking of ordering alexa488nm-phalloiden, alexa568- streptavidin, and syto60 from molecular probes to start getting used to the scope and the lasersharp bio-rad software. The immunohistotech in the lab uses an ABC dtection system with Dab. The first probe should contrast the cytoskeleton well(stain F-actin), the second probe will label any primary Ab, and the third will bind to DNA and contrast the nucleus. Will using these three probes together be a good starting point to becoming adept at labeling and producing high quality images on the LSCM including optical sectioning and 3D (and 4D sometime in the future)? I welcome suggestions Thank you
http://www.resolve3d.com/in_the_news_resview.html Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
Hi Listers, I have a BioRad Confocal MRC1000 on a Nikon Optiphot (upright scope) and would like to use it on an inverted microscope but the only thing I have is an older Diaphot with the 80/20 prism. Of course this is unacceptable as there is no shutter to prevent someone from inadvertently looking into the eyepieces while scanning. Does anyone know if and how the sliders on the Diaphot can be modified so as to serve as a shutter, i.e. in one position the scanner input is blocked to allow viewing and in another it is directed only to the objective/specimen? I assume it would mean a modification to the internal prisms. Has anyone done this? Source for parts? Procedure? Does anyone do this as a business? I called my local Nikon dealer and was told that the 80/20 can not be converted. Would I be better off to by a new inverted scope? Issues: cost, new infinity corrected optics. What are the issues with this changeover if you have done this. If someone who is on the confocal list could forward this, I would appreciate it. I have not been able to sign on since I changed my email address. Thanks so much for advice and help. Damian Neuberger, PhD Senior Research Scientist Baxter Healthcare Corp. email: damian_neuberger-at-baxter.com Fax: 847/270-5897Phone: 847/270-5888
Along this thread, when we print B&W images on a HP deskjet, we set printer properties to Grayscale, yet when we examine the image with a hand lens, we can see color dots. Why? (perhaps I missed the answer in a previous recent message, sorry). Damian
I got a question from one of the Professors in Pathology here in Tromso, and she wants to know if there are anyone selling seminar boxes with specimens (Light microsopy)from human pathology/anatomy?
Best regards Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso N-9037 TROMSO Norway
I'm not certain but your Sorvall unit may be similar in design to the Polaron CPD units that we service and sell in the U.S. If you could send us a digital photograph and/or copies of any information on the unit that may still be around, we will be happy to do some research to see if we can help you.
Please feel free to contact me off line at mnesta-at-ebsciences.com or by phone at (800) 992-9037.
Mike
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: swiding [mailto:swiding-at-astro.temple.edu] Sent: Wednesday, June 20, 2001 9:38 AM To: microscopy-at-sparc5.microscopy.com
Hello List,
I am trying to repair our Sorvall critical point dryer. I would like to replace the needle valve assemblies as they are starting to leak. Does anyone know if parts are available for this system? If not, we will have to get it replaced.
We just had a demo of their system from the local sales person, and I must say I was greatly impressed. The company is very young and full of some great ideas. The sales crew seems eager to work with potential customers. The demo was good enough for the powers that be here to approve some "proof of concept" work with some of our own samples. The software is simply amazing in its potential. It seems to have a very user friendly, and intuitive interface that is quite powerful. Obviously the biggest limitation is resolution. Despite the claims of "high resolution", they are limited to typical light microscopy resolutions of about 0.2um/voxel. Another limitation is your material. If you are talking about biological samples, there is no problem, but currently certain materials do present a problem for them.
If you have more specific questions about what I saw at the demo, please feel free to contact me.
Sincerely,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
sterling stoudenmire To: MICROSCOPY-at-sparc5.microscopy.com {sstouden-at-the cc: links.com} Subject: does anyone have a resolve workstation
06/21/2001 06:42 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
if so, how well is it liked?
http://www.resolve3d.com/in_the_news_resview.html Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
I'm not certain but your Sorvall unit may be similar in design to the Polaron CPD units that we service and sell in the U.S. If you could send us a digital photograph and/or copies of any information on the unit that may still be around, we will be happy to do some research to see if we can help you.
Please feel free to contact me off line at mnesta-at-ebsciences.com or by phone at (800) 992-9037.
Mike
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: swiding [mailto:swiding-at-astro.temple.edu] Sent: Wednesday, June 20, 2001 9:38 AM To: microscopy-at-sparc5.microscopy.com
Hello List,
I am trying to repair our Sorvall critical point dryer. I would like to replace the needle valve assemblies as they are starting to leak. Does anyone know if parts are available for this system? If not, we will have to get it replaced.
Boeckler/Ventana/RMC/Dupont-Sorvall no longer provide on-site service for old MT2-B microtome, and no support for our MT-5000 microtome. Does anyone know of a good, independent service provider for these instruments in the SE United States?
Julie Piraino Laboratory Manager Smithsonian Maine Station 701 Seaway Drive Fort Pierce, FL 34949 561-465-6630 x143 fax 561-461-8154
I am using the threshold feature of NIH Image to analyze images of cell cultures of individual neurons with processes. We are attempting to quantitate the number of processes in each culture. However, when the threshold function is applied, if a polygon is formed by the processes, NIH Image fills in the space between the "lines" (coverts all the pixels to black = "above threshold"). Since we are trying to measure the number of pixels above threshold, the filled space is counted as part of the number, thus preventing an accurate measurement.
Is there a way to stop the "fill" from occurring?
Thanks.
Paul Madtes Jr. pmadtes-at-mvnc.edu Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
Dear Listers, Does anyone have experience in isolating biofilms from sediment prior to fixation. I thinking of doing a pretreatment with Tween-20 but would appreciate suggestions from anyone who has prepared films for imaging in the LM and SEM. Rosemary -- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please comment on reliability, ease-of -use, lighting, etc. for those machines. Our lab is going to purchase a used machine in July.
I found that the learning curve was greatly improved by starting with another Molecular Probe product. I acquired the 6um FocalCheck beads (I got F-14807)(Search MP for document MP 07234) that fit our system - they should be fairly generic. I have made up my own slides. You will probably want one with three colors. The idea is to determine through experience the relationship between Mag Power and Optical Section Thickness. The problem in CSLM is not the x-y plane, it is learning how to get the z-axis right, and how to balance the channels. Choosing the other probes is a question that can only be answered by knowing as much as possible about the spectral characteristics of the excitation and emission filters in your system so that you can minimize crosstalk. NOTE: There is a paucity of information about filters that forces us to be empiric in setting up experiments, at least that's been my experience so far. I keep on asking for the information from the system vendor.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: "woxberry-at-downstate.edu"-at-sparc5.microscopy.com } Sent: Wednesday, June 20, 2001 8:04 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: New Confocal user } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am a long time Tem person starting a new job doing both TEM and } LSCM. Its very exciting learning LSCM but I need some suggestions for } ordering probes. Our lab has a Kr/Ar laser and will be doing } neuropathology research, mostly with fixed human brain samples, but } also tissue culture cells both live and fixed. A vibratome is in our } future. I was thinking of ordering alexa488nm-phalloiden, alexa568- } streptavidin, and syto60 from molecular probes to start getting used to } the scope and the lasersharp bio-rad software. The immunohistotech in } the lab uses an ABC dtection system with Dab. The first probe should } contrast the cytoskeleton well(stain F-actin), the second probe will } label any primary Ab, and the third will bind to DNA and contrast the } nucleus. Will using these three probes together be a good starting } point to becoming adept at labeling and producing high quality images } on the LSCM including optical sectioning and 3D (and 4D sometime in the } future)? } I welcome suggestions } Thank you } } Bill } }
Hello All, I have a Jeol 5900 SEM, and I would like to purchase a BSE detector for it. Does anyone in the group have any suggestions/recommendations/warnings? Is it pretty straight forward to purchase a third-party detector and get it to work with our microscope, or is it best to go through the vendor? Thank you for any input that you can provide.
Brad Johnson Pacific Northwest National Lab P.O. Box 99, K6-24 Richland, WA 99352 voice: 509-372-1635 fax: 509-376-3108
I found that the learning curve was greatly improved by starting with another Molecular Probe product. I acquired the 6um FocalCheck beads (I got F-14807)(Search MP for document MP 07234) that fit our system - they should be fairly generic. I have made up my own slides. You will probably want one with three colors. The idea is to determine through experience the relationship between Mag Power and Optical Section Thickness. The problem in CSLM is not the x-y plane, it is learning how to get the z-axis right, and how to balance the channels. Choosing the other probes is a question that can only be answered by knowing as much as possible about the spectral characteristics of the excitation and emission filters in your system so that you can minimize crosstalk. NOTE: There is a paucity of information about filters that forces us to be empiric in setting up experiments, at least that's been my experience so far. I keep on asking for the information from the system vendor.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: "woxberry-at-downstate.edu"-at-sparc5.microscopy.com } Sent: Wednesday, June 20, 2001 8:04 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: New Confocal user } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am a long time Tem person starting a new job doing both TEM and } LSCM. Its very exciting learning LSCM but I need some suggestions for } ordering probes. Our lab has a Kr/Ar laser and will be doing } neuropathology research, mostly with fixed human brain samples, but } also tissue culture cells both live and fixed. A vibratome is in our } future. I was thinking of ordering alexa488nm-phalloiden, alexa568- } streptavidin, and syto60 from molecular probes to start getting used to } the scope and the lasersharp bio-rad software. The immunohistotech in } the lab uses an ABC dtection system with Dab. The first probe should } contrast the cytoskeleton well(stain F-actin), the second probe will } label any primary Ab, and the third will bind to DNA and contrast the } nucleus. Will using these three probes together be a good starting } point to becoming adept at labeling and producing high quality images } on the LSCM including optical sectioning and 3D (and 4D sometime in the } future)? } I welcome suggestions } Thank you } } Bill } }
I work with ultrasmall nanosized particles, as small as 2 nm, with much lower contrast than metal particles such as gold and silver. These ultrasmall particles don't show up very well on TEM, even with optimal TEM settings. I thought what would help to increase the contrast dramatically would be to use a TEM grid that has very low background signal. I have tried grids with ultrathin carbon film (about 5 nm thick) but the background signal is still too high for these particles to show up clearly. Holey carbon grids is an option except they don't give me enough particles for particle sizing. Any suggestions would be very much appreciated.
In a recent issue of "PEI", May, 2001 (http://www.peimag.com) the following sampling of Inkset Resources was offered in an article entitled: "Inksets for Desktop Inkjet Printers", by Theano Nikitas.
The point of the article was the difference between pigment and dye inks, and those that are specially formulated for archival prints.
I am only using those that are not Printer Manufacturers
1. Cone Editions Press: http://www.inkjetmall.com
2. Luminos Photo Corp: http://www.luminos.com
3. Lyson USA Inc.: http://www.lysonusa.com
4. MIS Associates Inc: http://inksupply.com
5. MediaStreet.com: http://mediastreet.com
6. Xtreme gamut: http://xtremegamut.com
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Paula Allan-Wojtas } Sent: Wednesday, June 20, 2001 7:47 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Printers and paper } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, all, } } This question has come up for us at a good time, with this ongoing } discussion of printers. } } We recently purchased an Epson 880 ink jet printer for our EM/LM lab, and } are very happy with it. The black and white prints look excellent. We have } tried a number of papers, and find that the Epson "Photo Quality Glossy } Paper" and "Photo Paper" give the best results for our purposes. } } We are wondering how to go about making our prints more permanent from } these printers. If the prints have just been printed and aren't dry yet, } or if they are dry and are in contact with any moisture, they smudge quite } easily. Is there any coating - lamination or spray, or something else } which we can use to protect the surface? } } In connection with this, we have also found, especially when we have } printed b/w prints, that if we leave them in a room with full spectrum } fluorescent lights for a few days, that they begin to change colour, and } start to take on a reddish tinge. The instructions that came with the } paper say to not leave prints exposed to sunlight - but we're talking } about room lights, here! Has anyone else found this happens to them, and } is there any way of preventing/minimizing this effect without storing } prints in a light tight bag???? } } Any help or suggestions would be much appreciated. As always, thanks for } your help in advance. } } Regards, } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca } } }
I checked out there system from a general interest, "this looks cool" viewpoint. I recall the representative telling me that the technique works by serial sections. I also seem to recall that the images are taken of the remaining block rather than the removed sections.
It sounds like an interesting approach, and it will certainly have some applications. I did not see any immediate applications in my research. I was also unsure what kind of ultimate resolution could be achieved. It seems that the technique would be as limited by the thickness of the sections in the z-direction as it would by microscopy in the x- and y-directions.
At 09:26 AM 6/21/2001 -0400, you wrote: } Hi Sterling, } } We just had a demo of their system from the local sales person, and I must } say I was greatly impressed. The company is very young and full of some } great ideas. The sales crew seems eager to work with potential customers. } The demo was good enough for the powers that be here to approve some "proof } of concept" work with some of our own samples. The software is simply } amazing in its potential. It seems to have a very user friendly, and } intuitive interface that is quite powerful. Obviously the biggest } limitation is resolution. Despite the claims of "high resolution", they } are limited to typical light microscopy resolutions of about 0.2um/voxel. } Another limitation is your material. If you are talking about biological } samples, there is no problem, but currently certain materials do present a } problem for them. } } If you have more specific questions about what I saw at the demo, please } feel free to contact me. } } Sincerely, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 } } } From: sterling stoudenmire
In my point of view, it's almost impossible to have gray tones with RGB (3 color scheme) color cartridges. CYMK (4 color) will do right if printer set for gray-scale (to use black cartridge only). To simplify my life I removed color cartridges and replaced them on black one for B&W prints. For may model it's possible to use big black cartridge instead color assembly. I always replace color ribbon on black one in our Tektronix Dye-sub (actually, reverse: black ribbon installed by default and we replace that on color if necessary).
Sergey
At 07:22 AM 6/21/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
At 12:53 PM 6/21/2001 -0500, Warren E Straszheim wrote:
I also looked into this a while back, it's an interesting approach. Essentially the sample is fluorescently stained with something like eosin and embedded in an opaque plastic. The surface is imaged, a micron or two is shaved off, the new surface is imaged, and so on. This gives you a (huge) 3D data set that can be viewed at various magnifications and resolutions. You lease/buy a computer system from them, send them the sample and they send you a DVD with the data. The software is quite fast and easy to use. I haven't used their services yet, am still waiting for an appropriate sample.
Tom
} I checked out there system from a general interest, "this looks cool" } viewpoint. I recall the representative telling me that the technique works } by serial sections. I also seem to recall that the images are taken of the } remaining block rather than the removed sections. } } It sounds like an interesting approach, and it will certainly have some } applications. I did not see any immediate applications in my research. I } was also unsure what kind of ultimate resolution could be achieved. It } seems that the technique would be as limited by the thickness of the } sections in the z-direction as it would by microscopy in the x- and } y-directions.
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
I guess my question would be "what is the investigator's definition of a biofilm?" If you detergent-treat sediment, you are likely to remove a substantial majority of the attached bacterial biomass from suspended solids, but this will also contain non-biofilm (porewater) cells and the majority of the spatial information in the sample (relationships of cells to substrata, and of cells to cells) will be lost. Why do you want to "isolate" the biofilms? Why not examine them directly (you are using microscopy - virtually the ONLY way to examine biofilms directly).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
I have used GW Electronics BSE systems for many years. I purchased my last system just a few years back. With both the olde and new systems, I had excellent results -performance is great. The equipment is very well made and has been extremely reliable. I have no connection with GW other than being an experineced and satisfied user.
GW can interface to just about any scope.
Woody White McDermott Technology, Inc. http://woody.white.home.att.net
} -----Original Message----- } From: Johnson, Bradley R [mailto:Bradley.Johnson-at-pnl.gov] } Sent: Thursday, June 21, 2001 11:54 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: SEM: Recommendations for BSE detectors } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a } BSE detector for } it. Does anyone in the group have any } suggestions/recommendations/warnings? Is } it pretty straight forward to purchase a third-party detector } and get it to work } with our microscope, or is it best to go through the vendor? } Thank you for any } input that you can provide. } } } Brad Johnson } Pacific Northwest National Lab } P.O. Box 99, K6-24 } Richland, WA 99352 } voice: 509-372-1635 } fax: 509-376-3108 } } Bradley.Johnson-at-pnl.gov }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please } comment on reliability, ease-of -use, lighting, etc. for those } machines. Our lab is going to purchase a used machine in July. }
We have had an Ultracut E for 15 years with only occasional and relativley minor repairs needed. When we have needed service, Tek-Net of Lakewood, NJ has been excellent.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
} Date: Thu, 21 Jun 2001 14:22:08 -0700 } To: "Thearith H. Ung" {tung-at-qdots.com} } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: TEM-Ultrathin film grid } } Dear Thearith } } 5 nm carbon film is very thick. I am using 1.5-1.8 nm carbon film } on carbon 2 mkm holey film for 1.4 nm for NanoGold particles. You could } also try dark-field mode to enhance the contrast. It's also possible to } decrease the carbon film thickness because it's so strong. I measure the } actual carbon thickness by TM-450 MAXTEK thickness monitor (0.01 nm } resolution). I am using my own design "electron gun" and oil-free vacuum } apparatus for carbon film preparation. } } Sergey } } At 09:51 AM 6/21/01 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Try Bob Ruscica at ETP-USA. He handles Robinson scintillator detectors. Probably the best type made. Very sensitive, versatile and reliable. Not high cost, either. They can be retrofitted to most any scope.
gary g. (highly satisfied Robinson BSE user)
At 08:54 AM 6/21/2001, you wrote:
} Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a BSE } detector for } it. Does anyone in the group have any } suggestions/recommendations/warnings? Is } it pretty straight forward to purchase a third-party detector and get it } to work } with our microscope, or is it best to go through the vendor? Thank you } for any } input that you can provide. } } } Brad Johnson } Pacific Northwest National Lab } P.O. Box 99, K6-24 } Richland, WA 99352 } voice: 509-372-1635 } fax: 509-376-3108 } } Bradley.Johnson-at-pnl.gov
It should works great, thanks for idea. Basically, for my B&W job I am using Tektronix dye-sub which performs fine. Sergey
At 07:13 PM 6/21/01 -0400, you wrote: } look for Quad-Black - it is a four or six grey shade ink set to replace } the color ones - the prints have more grey levels than film. } } } At 03:01 PM 6/21/01, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used an RMC MT-6000 XL for five years and I am using an Reichert Ultracut E since a couple of weeks. After getting used to the Reichert I like it a lot and think overall it is the better machine. It has some nice features, for example this integral water pump and the specimen trans-illumination etc. and I think the mechanics are more robust. One thing is better about the RMC MT-6000 XL. You can change the angle of the microscope and by that always get the best reflection of the water surface in the knife boat (independent from how high the water level is). The RMC is really not bad but I think a lot of details make the Reichert better.
Hi Craig, As you probably know the machines you are asking about definitely have some age on them. The lab here has a RMC MT-6000XL and an Ultracut E. I really like the Ultracut - it works like a charm and has lighting in every possible direction. I really dislike the design of the RMC MT-6000 and the lighting is either on above or below but not both unless you get a switch installed (we had the switch installed and it makes a big difference to me). The newer RMC ultramicrotomes are very nice but the Ultracuts are still the best ultramicrotomes. Can't comment on the Nova. best regards, Beth
} Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please } comment on reliability, ease-of -use, lighting, etc. for those } machines. Our lab is going to purchase a used machine in July.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
For the more frugal try Johnson's Paste Wax. It works on some inks and papers, not all. It does offer some protection against UV. It does protect against moisture, finger prints, dust and scratches. It does change the finish some.
If the wax causes the ink to smear a fixative spray will usually stop that.
Test it for several weeks before putting it on an expensive display.
Wax also works on regular photographs. The way I discovered it was on B&W prints that were displayed outdoors in the weather for several months.
Gordon
Gordon Couger Stillwater, OK
.From: "George Laing" {scisales-at-ngscorp.com}
Luminos http://www.lumijet.com has a spray called "Imagesheild" to protect prints. $18/ can. Quoting their website: "LUMIJET IMAGESHIELD will become your favorite print protector. It's a convenient ozone-friendly low-odor aerosol spray that significantly improves wet-fastness. In addition to protecting delicate inkjet images from moisture, it also shields images from fingerprints and harmful UV rays. This unique product will not only help extend useful print life, it is totally transparent; preserving the original print surface characteristics. From dead matte to glossy, or from canvas to silver foil, you'll never know it's there."
} We are wondering how to go about making our prints more permanent } from these printers. If the prints have just been printed and } aren't dry yet, or if they are dry and are in contact with any } moisture, they smudge quite easily. Is there any coating - } lamination or spray, or something else which we can use to } protect the surface?
There were recently (last 6-8 months) some issues with color shift on some Epson papers. Epson determined the shift was from high concentrations of ozone, and not due to exposure to light. Epson recommends Heavyweight Matte or Epson Photo paper for maximum lightfastness.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com } } In connection with this, we have also found, especially when we } have printed b/w prints, that if we leave them in a room with } full spectrum fluorescent lights for a few days, that they begin } to change colour, and start to take on a reddish tinge. The } instructions that came with the paper say to not leave prints } exposed to sunlight - but we're talking about room lights, here! } Has anyone else found this happens to them, and is there any way } of preventing/minimizing this effect without storing prints in a } light tight bag???? }
I have used a 'Super Nova' for 11 or 12 years and the only thing that has needed to be replaced/repaired was the fluorescent light source. We are starting to have a niggling problem when switching on the light - it can take several attempts and it looks like there might be some sort of dedicated controller to be fixed. The only other thing that may need to be replaced is the belt (motor drive band) which is beginning to slip when manual raise and lower is used on the specimen arm. It's never needed a service engineer though. The reason we/I opted for the LKB was that I had used them for years and liked their reliability - there is no thin section mechanical feed to go wrong. The 'Super Nova' was a later model than the Nova and has a completely sealed control panel which keeps the dust and probably water out. It can also maintain specimen position accurately when you turn the thermal feed off and the cut stroke is powered rather than just relying on gravity. I don't know if these are standard on the Nova. The Ultracut E has nicer optics, specimen adjustment is superb and it is much less sensitive to external vibration but I cannot swear to it's long term use because I've only used it in demonstrations. I really think it would be worthwhile if you can get a demonstration (especially if you can take a specimen to cut) because these machines are all very good and it may come down to personal taste. One thing you might to want to examine is the availability of spares, accessories and servicing for each machine - in the UK it is becoming difficult to get spares for the 'Super Nova'.
I hope this helps.
The usual disclaimers apply: my opinions etc. and no affiliations to any company.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
craig klotz wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please } comment on reliability, ease-of -use, lighting, etc. for those } machines. Our lab is going to purchase a used machine in July. } } TIA, } Craig
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Hi all, We had a short in the objective lens coil a while ago and it was changed. What was happening was that we lost all control of the beam position and focus. We usually work at 15 kV (suitable for most of our needs), but now need to do something at 25. However, when we increase to this acc. voltage, we are finding a similar problem - beam pops out of focus (goes big and diffuse) and shoots off somewhere. We can get it back by pressing the Obj. Pol. button. This normally works if we get the same thing at 20 kV and less, but not at 25. The effect only occurs once an hour at 22 kV. Anyone have an idea why this is happening? By the way, for those who would undoubtably ask, the I is 100 nA for what we are at the the moment. Comments? Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (try your luck!) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
Dear Bradley, I have used the JEOL BSE detector, the GW BSE detector and the Robinson BSE detector and they all work well and I have had no reliability problems or interface problems. For the third-party detectors, the external signal just goes in through the "EXTERNAL" signal input on the SEM. At 08:54 AM 06/21/2001 -0700, you wrote: } } Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a BSE detector for } it. Does anyone in the group have any suggestions/recommendations/warnings? Is } it pretty straight forward to purchase a third-party detector and get it to work } with our microscope, or is it best to go through the vendor? Thank you for any } input that you can provide. } } } Brad Johnson Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Jim is right in what he says about the compatibility issues with Oxford/Link detectors and the restore signal being generated by the pulse processor and not the pre-amp as in most detectors. However we are reps for Thomson Scientific Instruments' WINEDS x-ray analyzers and pulse processing electronics in North America. TSI has the DX3000 bias and PX9000 Pulse processor that will interface to the Oxford/Link detectors. We have used it with both the older type and with the Pentafet type without any changes to the detector preamp. The PX9000 is a universal design and can be used with other manufacturers analyzers.
Douglas Connors General Manager TNAS Inc. 7897 Hwy 19, Dane WI 53529 (608)798-2005 voice (608)798-1675 fax tnas1-at-msn.com ----- Original Message ----- } From: "Jim Nicolino" {JNicolino-at-xraydetectors.com} To: "Michael L. Boucher" {mboucher-at-tc.umn.edu} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 19, 2001 10:30 AM
If you are only imaging the particles, not in context of a particular environment, you can negative stain e.g. with phosphotungstic acid.
Sounds like you're not considering staining them but if the particles are reactive then that is a possibility too.
Richard
} } } "Thearith H. Ung" {tung-at-qdots.com} 6/21/01 9:51:50 AM } } }
Dear colleagues,
I work with ultrasmall nanosized particles, as small as 2 nm, with much lower contrast than metal particles such as gold and silver. These ultrasmall particles don't show up very well on TEM, even with optimal TEM settings. I thought what would help to increase the contrast dramatically would be to use a TEM grid that has very low background signal. I have tried grids with ultrathin carbon film (about 5 nm thick) but the background signal is still too high for these particles to show up clearly. Holey carbon grids is an option except they don't give me enough particles for particle sizing. Any suggestions would be very much appreciated.
I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) buffered fixative. I need to store the samples for a year or more - are either of these fixatives appropriate for long-term storage, and if not, what are the problems (e.g., tissue hardening?). Does anyone have suggestions for appropriate storage solutions, or have an idea of how long is OK in either of these fixatives? The samples in paraformaldehyde are used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. Thanks in advance for your experience/advice.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Thanks for the suggestion. I thought about staining as well, except that these particles tend to get etched by acids. However, it might still work if only a small amount of phosphotunstic acid is used. So it is definitely worthwhile looking into it.
Thearith
-----Original Message----- } From: Richard Thrift [mailto:Richard_Thrift-at-skyepharma.com] Sent: Friday, June 22, 2001 9:57 AM To: tung-at-qdots.com; Microscopy-at-sparc5.microscopy.com
Thearith,
Have you got a possibility to do STEM ? If there are some high Z atoms in your samples, you should get enough contrast to see the particles.
Andreas
} Dear colleagues, } } I work with ultrasmall nanosized particles, as small as 2 nm, with much } lower contrast than metal particles such as gold and silver. These } ultrasmall particles don't show up very well on TEM, even with optimal TEM } settings. I thought what would help to increase the contrast dramatically } would be to use a TEM grid that has very low background signal. I have tried } grids with ultrathin carbon film (about 5 nm thick) but the background } signal is still too high for these particles to show up clearly. Holey } carbon grids is an option except they don't give me enough particles for } particle sizing. Any suggestions would be very much appreciated. } } } Regards, } } Thearith } } } }
Sounds like a contact problem with the OL polarity switch or relay. At higher KV's the lenses need higher current. 22 KV is probably the limit for the contact to cut-out.
Regards,
Earl
----- Original Message ----- } From: "Dr Malcolm Roberts" {m.roberts-at-ru.ac.za} To: "Microscopy discussion group" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, June 22, 2001 6:09 AM
Hi Rachel...
I would imagine that storage in 70% ethanol might be a better bet... brought up through an alcohol series. This is a standard practice for long term storage of museum specimens after fixation, and for most EM labs.
Check out following for reference: Simmons, J.E. 1995. Storage in Fluid Perservatives. In Storage of Natural History Collections: A Preventative Conservation Approach. Vol. 1. C.L. Rose, C.A. Hawks, and H.H. Genoways, eds. Society for the Preservation of Natural History Collections, Iowa City. 161-181.
Best regards,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Fri, 22 Jun 2001, Rachel Spicer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Richel
If you want to store for long term then better preserve in a deep freezer at -20 to -30 deg as it is and when you want to proceed, allow it to come down at 4 deg and proceed as per normal procedure.
Second option is fix in a fixative and then store in washing buffer at 4 deg celcius, but keep one thing is in mind that you should keep changing this buffer after a short time span, say 15-20 day. Your tissue well perserve in this condition. Like this i have preserved the animal tissues in best of condition for three months.
If you try to preserve in fixattive then their is a chacnes of hardening the material or if you preserves in alchohol 70%, the chances of shrinkage is more.
for best prservation methods Ref. the book by M.A. Hayat, Techniques in Electron Microscopy.
With Best Regards Arti Harle
Angela Klaus {avklaus-at-amnh.org} wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Rachel...
I would imagine that storage in 70% ethanol might be a better bet... brought up through an alcohol series. This is a standard practice for long term storage of museum specimens after fixation, and for most EM labs.
Check out following for reference: Simmons, J.E. 1995. Storage in Fluid Perservatives. In Storage of Natural History Collections: A Preventative Conservation Approach. Vol. 1. C.L. Rose, C.A. Hawks, and H.H. Genoways, eds. Society for the Preservation of Natural History Collections, Iowa City. 161-181.
Best regards,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Fri, 22 Jun 2001, Rachel Spicer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
Ms. Arti Harle Regional Sophisticated Instrumentation Center Indian Institute of Technology-Bombay (IIT-B) Powai, Mumbai - 400076. India Phone : 91-22-5767691 Extn 7694 91-22-5720109/ 5721039 (Resi) E-mail : aartih-at-usa.net
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Thearith H. Ung wrote the following: ========================================= I work with ultrasmall nanosized particles, as small as 2 nm, with much lower contrast than metal particles such as gold and silver. These ultrasmall particles don't show up very well on TEM, even with optimal TEM settings. I thought what would help to increase the contrast dramatically would be to use a TEM grid that has very low background signal. I have tried grids with ultrathin carbon film (about 5 nm thick) but the background signal is still too high for these particles to show up clearly. Holey carbon grids is an option except they don't give me enough particles for particle sizing. Any suggestions would be very much appreciated. =============================================== This has become quite a generic problem for many who are working with nano size particles, and many have found the "solution" to be the use of silicon nitride membrane window grids that have been offered by SPI Supplies for several years. Information about these grids can be found on URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
The silicon nitride is completely structureless and featureless and seems to provide excellent electron transparency. The other advantage is that they are sufficiently robust to survive temperatures in excess of 1000° C. And there is no carbon grain to be confused with the nanoparticles of interest.
Disclaimer: SPI Supplies offers silicon nitride membrane window grids for this type of application.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
There are other negative staining compounds available - ammonium molybdate and uranyl acetate for instance. However, my first try would be shadow-casting, by evaporating some Pt/C at about 10 degrees to the specimen. Cheers Jim Darley
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On Saturday, June 23, 2001 6:06 AM, Thearith H. Ung [SMTP:tung-at-qdots.com] wrote: } } } } Hi Richard, } } Thanks for the suggestion. I thought about staining as well, except that } these particles tend to get etched by acids. However, it might still work if } only a small amount of phosphotunstic acid is used. So it is definitely } worthwhile looking into it. } } Thearith } } -----Original Message----- } } From: Richard Thrift [mailto:Richard_Thrift-at-skyepharma.com] } Sent: Friday, June 22, 2001 9:57 AM } To: tung-at-qdots.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: TEM-Ultrathin film grid } } } If you are only imaging the particles, not in context of a particular } environment, you can negative stain e.g. with phosphotungstic acid. } } Sounds like you're not considering staining them but if the particles are } reactive then that is a possibility too. } } Richard
Readers, Kindly allow this short reminder that again at the M&M 2001 Conference we will hold a "Just For Fun Micrograph" Contest. The concept being composite images, each of two or more images, one of which must be microscopical in nature. $600 cash in prizes. Contact me direct and I will provide full detail. Don Grimes, Microscopy Today
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3. CUSTOMER LETTERS can be created from the Quotation and Sales Order sections.
4. SHOP TRAVELER/WORK ORDER. Once a Sales Order is accepted, the sales order information can be transformed into a shop traveler/work order on a "point and click" basis. Each item on the Sales Order becomes a shop traveler/work order, with each step of production of the item then listed on the traveler/work order. Each such traveler/work order is tied back into the Sales Order. The shop traveler/work order allows for the entry of line items, and notes on each line item. The shop traveler/work order contains a "notes" section. The Shop traveler/work order allows for the storing or attachment of drawings to the traveler/work order. The shop traveler/work order also contains a "drop down," from which standard processes can be selected for inclusion on the shop traveler/work order. The shop traveler/work order numbers progress in order of production sequence, and re numbers them if new steps are added. The shop traveler/work order allows for change orders or revisions, and numbers changes in sequence of he original shop traveler/work order number; i.e., 100, 100-1, 100-2, etc. All shop traveler/work orders and related revisions are retained in memory for future reference. The shop traveler/work order is bar coded for tracking of production step by step, and production of ongoing client status reports. Bar coding includes the ability for an employee to "swipe" their own ID bar code for recording in the system as to who upgraded what step. The shop traveler/work order function also allows for manual update of production status. The shop traveler/work order allows for quality control sign off, and the final production of certifications, either from a "canned" list, or hand typed in on a case by case basis.
5. INVENTORY. The application includes an inventory section, which allows operations to check materials inventory in and out. The inventory section allows for the comparison of inventory received against a P.O., and produces an "overage/underage" report of inventory received as compared against the P.O. The inventory section allows for the setting of minimum (re-order now!) and maximum inventory amounts, and produces reports showing what inventory needs to be ordered, as well as inventory that is at or above the maximum set to have in house. The inventory section also tracks "partially shipped" orders, which are tied in to the shipping function. This section shows how much completed product under a particular order has been actually shipped to a client, and how much remains to be shipped. The balance is adjusted as shipments are made.
6. REQUEST FOR PURCHASE. The application allows operators to produce a Request For Purchase for accounting for any inventory items, which need to be ordered. Inventory items have a drop down of approved vendors for each item.
7. REQUEST FOR BID. The application allows operators to produce a Request For Bid for accounting to send to Vendors for any inventory items, which need to be ordered. Inventory items have a drop down of approved vendors for each item to which Requests For Bid can be sent.
8. INVOICE. The application produces an invoice/invoice detail for all completed items ready to be billed/shipped to clients.
9. PRODUCTION OUTPUT STATUS. The application produces a date range selectable report on how much product, and the value of the product, which was completed during a selected date range. The application also produces a report on how many orders, and the value of those orders, which remain to be completed during a selected date range.
10. The application produces SHIPPING DOCUMENTS as per selected shippers, and produces a PACKING SLIP.
11. The application has a "FIND" FUNCTION in selected sections, allowing for searches by customer name, work order number, etc.
12. The application has "AUTO FILL;" i.e., when an operator starts to type in a name, number, etc. all related information auto fills after the first few letters or numbers are typed in.
Job Master is currently being sold in the marketplace for $2,495.00 per package. However, if we receive your order by June 22th, your total price will be $1,495.00.
Again, if you have any questions at all, or would like to place your order, please call me on my direct line, (661) 254-9926. Thank you!
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This has been discussed before, but... If anyone has a summary regarding contract services that they could share, I would appreciate it. (They seem to be lower priced than electon microscope vendors.) Any recent experience positive or negative with contract services, particularly 1) Advanced Research Systems (Alan Sampson) in Illinois or 2) Sci. Inst. & Applications (V. Feingold) of Duluth, GA or 3) Other EM service providers covering the Midwest. Thanks for your help.
Remember that paraformaldehyde is a reversible fixative, so it isn't appropriate to fix in pform then store in a buffer. I've stored animal tissue for years in pform with no problems, though I'm sure it's not recommended. As for pform/glut, storage after fixation in cacodylate buffer is good, as nothing seems to grow in cacodylate.
Diana
} } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel --
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
How about storing part of the sample in buffer with sodium azide to inhibit microorganisms.
Dave
On Fri, 22 Jun 2001 14:38:15 -0400 Rachel Spicer {spicer-at-oeb.harvard.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have found that there are a few cacodylate tolerant organisms. Nonetheless I usually take the risk.
Dave
On Mon, 25 Jun 2001 14:28:00 +1000 Diana van Driel {dianavd-at-eye.usyd.edu.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi Rachel, } } Remember that paraformaldehyde is a reversible fixative, so it isn't } appropriate to fix in pform then store in a buffer. I've stored } animal tissue for years in pform with no problems, though I'm sure } it's not recommended. As for pform/glut, storage after fixation in } cacodylate buffer is good, as nothing seems to grow in cacodylate. } } Diana } } } } } Hello, } } } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } } buffered fixative. I need to store the samples for a year or more - are } } either of these fixatives appropriate for long-term storage, and if not, } } what are the problems (e.g., tissue hardening?). Does anyone have } } suggestions for appropriate storage solutions, or have an idea of how long } } is OK in either of these fixatives? The samples in paraformaldehyde are } } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } } Thanks in advance for your experience/advice. } } } } Rachel } -- } } } } Diana van Driel } Department of Clinical Ophthalmology } Sydney University } GPO Box 4337 } Sydney NSW } AUSTRALIA 2001 } } Phone 61 2 938 27278/27395 } Mob 0412 165 075 } Fax 61 2 938 27318 }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Beverly Giammara wrote asking for comments about EM service providers, other than the manufacturers, that cover the Midwest. I would be very appreciative if anyone can offer similar information about EM Service providers that cover Florida.
Sincerely,
Jill
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
We are currently assessing whether or not to replace our old diffraction simulation software and would in this respect be happy to receive the advice of the microscopy community and vendors.
The software should have the following characteristics: * Electron diffraction simulation, kinematical as well as dynamical calculation. * Handle multiple reciprocal lattices in one pattern - that is - several single crystals with different crystallography, size, shape, and orientation relationships, as well as powder diffraction. * Accurate intensity and structure factor determination. * Evaluate double diffraction and crystal shape effects - streaking. * The software should also be able to simulate X-ray diffraction patterns: powder, Laue, Guinier. * Support of common crystallographic/diffraction database formats. * The software should run under Windows NT 4.5 and have a professionally designed GUI. No compiling and linking, and no programming should be necessary but the possibility of doing this (e.g. scripting, ActiveX, Plug-Ins etc.) is a very positive feature.
Sincerely, Paul Baggethun ========================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA (724) 337 1760 paul.baggethun-at-alcoa.com ==========================
Please forgive. My first rule of subjugation is: "They can only learn and do what they are taught!" Thus, one hopes that you will NOT take umbrage with what follows. However, your teachers, ......
As you are in such a "Divine" location, I am hesitant to suggest anything, especially in light of your use of such holy words of biology as "chunks" and "paraformaldehyde". I will, however, and only with great temerity and the deepest humility, suggest that you try to gain access to a tome of non-Harvardian origin entitled, "Principles and Techniques of Electron Microscopy"(3d ed.), Hayat, M.A.[he once worked in Newark, NJ!!!], CRC Press, Boca Raton, FL(1989), pp. 69-70 (sections entitled: "Simultaneous Fixation for Light and Electron Microscopy", and "Tissue Storage".) ISBN: 0-8493-7111-2. My humility is manifested for the divinity, but my temerity is evoked by the ignorance. However, Hayat begins one paragraph with the following sentence, "The size of the tissue specimen and the objective of the study primarily determine the type of the fixative solution to be used." He further mentions 4% formaldehyde and 1% glutaraldehyde in a buffer at pH 7.4 with an osmolality of ~200mo for "medium" pieces (~3mm) for longer times up to 12 months. Plants get specific treatment on pp. 72-73. There are other books, though I must admit, notwithstanding the efforts of Science Librarians, most of them have been 'lifted' by the new wave of dedicated students and others during the past 30 years. Try the 'older' members of the biology faculty - non-molecular, if possible, - and remember that HCHO has been used in some states to INDUCE fluorescence in tissues!
Again, PLEASE forgive. The last time I spoke to a student, a pre-med by-the-by, I didn't understand him either.
We old-timers are severely challenged, because we were trained to:
choose a vocational path to follow, ask questions after we had gone to the library, judge Timothy Leary as the short form of a donkey, understand women as a great mystery of the universe, leave the gloves off when dissecting, PLEASE understand, and NOT ask about the war, use slide rules (to do logs), planimiters (to do areas), and Leroy Lettering Sets (to label figures), understand the divine nature of (inspiration!) discovery take very deep breaths in the lab during days of dissection,
mix formaldehyde ("Formalin") and phenol to make good embalming fluid, squirt phenol on a sponge and wipe to disinfect lab benches in bacteriology, know that et cetera WAS Latin not Siamese(now Thai), take deep breaths in bacteriology lab, accumulate good books while they were relatively inexpensive, know that phenol was hygroscopic, cut sections from any tissue as long as it came from an unlabelled jar, label the jar, remember Latin and Greek roots (On Divinity Avenue, is that "dis-" as in "to separate" or dice-" as in "to chop up"?), do our income taxes on cards, tell students off, treat your fiancée' as the virgin you hoped she was, know the difference between formaldehyde and paraformaldehyde, keep the lab windows open during dissections to keep your eyes from burning, cultivate your wife as your best friend, ignore HIV and gHSV - no one had ever heard of them, be terrified of pregnancy before marriage (the greatest deterrent of STD's in the 20th century!), and know the difference between discovery and invention (a la, polymerase, la la, and the PCR, la la).
We were taught BY the organic chemists to pay very strict attention TO the organic chemists, because they were said to know most of the important secrets. The physical chemists, at the time, were busily unraveling the secrets of life - only one 'A' every five years among hundreds of chemists! Biochemists were chemists. Biologists cultured cells, developed vaccines, cut sections, referenced papers from the 1860's, didn't teach DNA until the early 1960's when protein genes were defeated at the polls, and, just now, may be getting - in another decade - to the point where the calculus may have a place in their courses. In the 1950's, or so went the legend, Harvard Medical School (Heaven's Gate?) sent representatives to Lehigh, where my teachers worked, to discover the secrets of chemical pedagogy that prepared LU graduates so well for the divine medical training some few received at the hands of Harvard's Professeurs! The secret pedagogical trick they discovered at Lehigh consisted of: "NO MERCY!", "NO SYMPATHY!" and "TOTAL RIGOR!", the only means ever devised to overcome the (utter) shame of being considered near to, but nonetheless, NOT, the equal of those "IVY" places.
If you didn't have fun, I wish you had. AND, before you store in buffer, think about it (extraction of fixative and other things) long and hard!
Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Rachel Spicer } Sent: Friday, June 22, 2001 2:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Q re: long term storage in fixative } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
Novascan Technologies has a great deal of experience in the area of biochemical AFM and I'm sure we can help you with your project We have been developing AFM products for several years, but our specialty is fabrication of AFM probes and surfaces with specific surface chemistries. These chemistries, for example, can be used to couple molecules to surfaces for imaging and/or molecular characterization. Please contact us at info-at-novascan.com or visit www.novascan.com
Best Wishes,
Raj
At 04:17 AM 06/20/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
********************************************** Dr. Raj Lartius, CEO NovaScan Technologies, Inc. Iowa State University Research Park 2501 North Loop Drive Ames, Iowa USA 50010
Email: rlartius-at-novascan.com Voice: 515-795-3164 Fax: 515-795-4414 ********************************************** "Innovative Tools to Explore the Microworld"
In our lab in Massachusetts, we have a Balzers CPD-020 that is in need of repair. The pressure guage (WIKA, 0-160 bar) is leaking, and we'd like to have it replaced. This unit was made in Switzerland, but the manufacturer has been out-of-business for many years. We'd appreciate input from Balzers CPD owners regarding sources for parts. Any comments concerning safety issues would be welcomed also. Many thanks in advance.
Does anybody have experience with High Resolution Imaging Plate Scanner for Electron Microscopy (http://www.ditabis.de). Instead of conventional EM-films, flexible photostimulable phosphor screens of the same size, the so-called Imaging Plates, are loaded into the TEM magazine. Are they as good as negatives?
Thanks, Julian
_______________________________ Julián Martínez Fernández Senior Research Associate MS 106-5 Ceramic Branch NASA Glenn Research Center Cleveland, OH 44135 Phone: 1-216-433-5512 Fax: 1-216-433-5544
You may find that only the JEOL one will properly fit in, I investigated a GW Electronics one for my JEOL 840, but they didn't have a size that would fit exactly. Mind you, I have a (JEOL) optical microscope fitted, and the BSE detector would have had to fit into a severely constrained space on the end of the OM.
I would ask JEOL first, I find that their prices are sometimes better than those from third-party suppliers anyway.
} } } Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a BSE detector } for } it. Does anyone in the group have any } suggestions/recommendations/warnings? Is it pretty straight forward } to purchase a third-party detector and get it to work with our } microscope, or is it best to go through the vendor? Thank you for } any input that you can provide. }
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello, We just inherited an older model (10 years old) model E-3 Electroscan Environmental SEM. I have some questions in its operating vacuum modes as it has been quite a bit picky since we've installed it.
On going from a standby vacuum state into either Hivac or Wet vacuum modes, it will often have an error of can't pump column EC2. Also, in Wet mode, it will not open valve V1, so you can never see the beam. I've read over the manuals, but I was wondering if anyone out there has some experience with this model ESEM. We are thinking that one of the diffusion pump heater elements needs to be changed. Other times we operate the unit, it will maintain a Hivac vacuum mode for several hours without difficulty.
Any advice or comments, appreciated, Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I did study this subject a few years ago and find that this technique is not well optimized for EM. For many reasons:
- cost of the system is similar to the EM digital camera - you require to load/unload Imaging Plates into the cassettes (you waste the time) - it takes from 2 to 5 min to scan one Image Plate (waste time) - I am not sure exactly, but seems to me, that you have to scan Image Plates soon after tacking picture, otherwise the image will deteriorated, and seems to me they are sensitive to light too. - you don't have a chance to adjust your image before recording (as it happens in case of digital cameras). -number of "shots" is limited and Image Plates are princely - Image Plates may be damaged during loading/unloading which may reduce declared number of use (a few thousand times I believe).
The good things about Image Plates are: - Image plate has more pixels than EM digital cameras, so the resolution is better. - you don't have to make any changes in the microscope. - the linearity and dynamic range is similar to the EM digital cameras - easy to use and less problem related to the equipment.
My conclusion was: technically, it's a great stuff, but it does not help in practice. You still need some help from technician to load/unload plates and scan them and you have to pay a 20-40$K for that beauty. Not so promising technology for TEM at least.
This information based on my "study" performed a few years ago and may not reflect modern improvements in this area.
Sergey.
At 03:11 PM 6/25/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I was talking about particular DITABIS system Julián was mentioning in her original message (with reference to the manufacturer).
All digital cameras including Gatans are approximately 10 times more sensitive than 4489 film. I know it for sure because just returned back from Gatan's demonstration. Image Plate should have very similar characteristics in terms of sensitivity, dynamic range and linearity as modern digital cameras. The best things about Image Plates it's their pixel size: 5000 x 4500 total pixels per plate. It's greater than in digital cameras: 1000x1000/2000 in most cases. It's still less than you could get from film with $800 scanner: 6400x4800 for 1600 optical dpi. BUT!!! You have LOAD/UNLOAD plates into cassettes manually. In the dark. Stack them and load into the scanner (so, you have to have space for the scanner in dark-room). Scanner will process them automatically (and at that time you could not use dark-room for regular things, light from scanner). Load them back into cassettes, degas magazine and load it into microscope. I don't remember exactly, but it seems to me the cost of one plate is something around $100. So, I have two magazines with 50 cassettes each: 100x100$ plus scanner plus salary for technician plus computer and storage media. Ok, I forgot to mention that you have to ERASE plates before use, also, using special eraser. I don't remember, how many plates you could ERASE in one run. May be it's automatic too. Scanning: 3.5 min per plate x 50= 3 h.
As I mentioned before, the technique is great, but not so helpful for regular day-to-day basis EM and does not save time so much. It works fine for molecular biology applications in "phospho-imagers" but people need to scan a few plates per day (even one a week) and they save a lot of time: 10-15 min exposure vs 5-10 days! I just don't see reason to switch to this technique to myself. If I'll have such money, I would buy digital camera. It's more practical solution from my point of view.
Sergey
At 01:36 AM 6/26/01 -0500, you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Sergey, } } I last saw the unit demonstrated at the EUREM meeting in Brno last summer. } It is not as you described it. I won't say it is better but overall it is } comparable to the Gatan product. } } What impressed me the most about it is that } } a) exposure times are comparable to what is now used for conventional film } (the Gatan unit has long exposure times) and } } b) while the scanning step (of the digital plates) takes some time, it does } it all automatically, so there is no babysitting with the scanner. You are } free to do other things including exposing more digital plates. } } They might have one at the MSA meeting in Long Beach. } } Chuck } SPI Supplies } -------- REPLY, Original message follows -------- } } } Date: Monday, 25-Jun-01 08:20 PM } } } } From: Ryazantsev, Sergey, Ph. D. \ Internet: (sryazant-at-ucla.edu) } } To: Julian Martinez-Fernandez \ Internet: } } (julian.martinez-fernandez-at-grc.na) } } To: MICROSCOPY BB \ Internet: } } (microscopy-at-sparc5.microscopy.com) } } } } Subject: Re: High Resolution Imaging Plate Scanner for Electron } Microscopy } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To } } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On- } Line } } Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I did study this subject a few years ago and find that this technique is } not } } well optimized for EM. For many reasons: } } } } - cost of the system is similar to the EM digital camera } } - you require to load/unload Imaging Plates into the cassettes (you waste } } the time) } } - it takes from 2 to 5 min to scan one Image Plate (waste time) } } - I am not sure exactly, but seems to me, that you have to scan Image } } Plates soon after tacking picture, otherwise the image will deteriorated, } } and seems to me they are sensitive to light too. } } - you don't have a chance to adjust your image before recording (as it } } happens in case of digital cameras). } } -number of "shots" is limited and Image Plates are princely } } - Image Plates may be damaged during loading/unloading which may reduce } } declared number of use (a few thousand times I believe). } } } } The good things about Image Plates are: } } - Image plate has more pixels than EM digital cameras, so the resolution } is } } better. } } - you don't have to make any changes in the microscope. } } - the linearity and dynamic range is similar to the EM digital cameras } } - easy to use and less problem related to the equipment. } } } } My conclusion was: technically, it's a great stuff, but it does not help } in } } practice. You still need some help from technician to load/unload plates } and } } scan them and you have to pay a 20-40$K for that beauty. Not so } promising } } technology for TEM at least. } } } } This information based on my "study" performed a few years ago and may not } } } reflect modern improvements in this area. } } } } Sergey. } } } } } } } } At 03:11 PM 6/25/01 -0400, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Does anybody have experience with High Resolution Imaging Plate Scanner } for } } } Electron Microscopy (http://www.ditabis.de). Instead of conventional } } } EM-films, flexible photostimulable phosphor screens of the same size, the } } } } so-called Imaging Plates, are loaded into the TEM magazine. Are they as } } } good as negatives? } } } } } } Thanks, } } } Julian } } } } } } _______________________________ } } } Julián Martínez Fernández } } } Senior Research Associate } } } MS 106-5 Ceramic Branch } } } NASA Glenn Research Center } } } Cleveland, OH 44135 } } } Phone: 1-216-433-5512 } } } Fax: 1-216-433-5544 } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } } } } -------- REPLY, End of original message --------
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I contacted ditabis several months ago. Beyond a confirmation email from the company, I haven't received any from them.
Steve Widing Temple University
On Mon, 25 Jun 2001, Julian Martinez-Fernandez wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anybody have experience with High Resolution Imaging Plate Scanner for } Electron Microscopy (http://www.ditabis.de). Instead of conventional } EM-films, flexible photostimulable phosphor screens of the same size, the } so-called Imaging Plates, are loaded into the TEM magazine. Are they as } good as negatives? } } Thanks, } Julian } } _______________________________ } Julián Martínez Fernández } Senior Research Associate } MS 106-5 Ceramic Branch } NASA Glenn Research Center } Cleveland, OH 44135 } Phone: 1-216-433-5512 } Fax: 1-216-433-5544 } }
For electron microscope service try Vitaly Feingold who is based out of Georgia. His references are excellent!! His phone number is 770-232-7785. E-mail address is: vitalylazar-at-worldnet.att.net
-----Original Message----- } From: Jill Verlander Reed [mailto:verlaj-at-medicine.ufl.edu] Sent: Monday, June 25, 2001 12:20 PM To: Microscopy-at-sparc5.microscopy.com
Fellow microscopists -
Beverly Giammara wrote asking for comments about EM service providers, other than the manufacturers, that cover the Midwest. I would be very appreciative if anyone can offer similar information about EM Service providers that cover Florida.
Sincerely,
Jill
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
Dear Rachel, After an entire evening of self-immolation, I have returned to the table to extend my apology for taking advantage of you yesterday. I really do apologize. I looked across the table at my own daughter last evening, and I realized that she might just have failed to see any humor at all in my response had she requested the same information. Having realized that possibility, I spent the rest of the evening in shock at my probable insensitivity. As an explanation, you received, in response to your entirely innocent query, a message composed of every quip I have wanted to make to my children, their teachers, their teachers' parents, my students, and probably a few I can't remember. I thought to make it humorous, and there are some who have decided that I succeeded, but I would suspect that you may have felt somewhat differently. Thus, this message in self-rebuttal. Not nearly as long, and hopefully more to the point. If you had walked into my office with the question you asked, I would have said, "Miss Spicer, we do not use words like "chunks" to describe specimens from which we wish to extract reasonable scientific information." You would have responded with some other nebulous term for the objects with which you were contesting, and I would have cajoled you, with a couple suggestions on precision in thinking and describing, to go to the library and start doing what you are obviously trying to learn how to do - namely, research. So, please let me begin again. My second rule is: "They only learn to think if they are forced." In that regard, I would have requested that you make a list of the steps in the process you are asked implement. For each step in such a process, there must be a purpose. For each step, there must be a prescribed outcome (I have come to despise that word!), and for each step, there must be BACKGROUND/HISTORY. My third rule is this. "The most difficult, and the most successful research for a bench scientist is done in the brain, the office, and/or the library." The work at the bench only certifies or debunks what was done before, but the same principal holds for even the most minor part of the process. You suggest that you must to store plant/wood tissue for a year. Most of us would want to know why? Why must you wait to at least embed. Why would you want to soak/extract the tissue for that length of time before acting on the NEXT step in the procedure. It is apparently not that you wish to archive the tissue/wood. So, with your list in your hand, the question for that step is, "Why?" "What is the purpose/reason for waiting?" I certainly cannot help you to progress in the trek to an answer until I know. Why? I'll give you an example. There was an undergraduate who accosted me in the hall during my first week as a graduate student. He asked me to point him in the direction of distilled water. I asked him what type he wanted. He assumed I was harassing him and told me to knock off the B.S. and just, "point!" I did. Unfortunately, his purpose was to subculture honey bee cells, and the 'distilled' water to which I pointed - the nearest - was made in a chromium plated metal still. Notwithstanding his rudeness, I should have insisted - through instruction, but I was fresh out of the military and I didn't have any time left for stupidity and self-destructiveness, Fourteen weeks later, he blamed me for the failure of his project - my first real interface with a baby boomer of the 'gimmee' type. I only felt sympathy for the instructor, but his advice (he had been in the military too) was to learn to take responsibility for the errors of students, for, someday I would be their teacher.
Over the past thirty years, what was a two-way street in learning when I was a student, has become one-way. I guess the ultimate point is this. If you are working on someone else's project, show him or her my notes. If you are working on a project of your own, and you are willing to exercise patience and learn, then when you ask for information from us - the MSA ListServer - you must provide us with specific information. I can promise you that you failed to receive many responses, because your question lacked three elements:
1. Precise terminology. 2. Purpose/Goal/Reason for need. 3. Trust.
Everyone on the ListServer has a job or values his/her time. Your request was informal in form, because it lacked 1) precise detail, 2) a reason for wanting to delay further processing (that would be important for any specific response), and 3) trust in us to be honest brokers and not steal your project or question its value. Far too many visitors to the Listserver ask a question as if in class, at the end of a lecture. We all came in late.
I hope you want to try again, if what little you have received has not helped. We would like to help as much as possible. We are here for that. Not for entertainment. I am here, because when I read the messages, I learn or have questions that I can address. I am here to be educated, just like you.
With closing apology for wordiness and philosophy, I am
Yours sincerely,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Rachel Spicer } Sent: Friday, June 22, 2001 2:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Q re: long term storage in fixative } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
Thanks to all who responded to my post re: long term storage of plant material in fixatives. The responses were as varied as the recommendations in the literature, but I find there is no substitute for hearing from those with first hand experience. If anyone is interested in the specifics I'd be happy to forward a collection of relevant responses. And finally, apologies for my misspelling of "Karnovsky". Thanks again.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Congratulations on your E3 acquisition - they're generally easy-running instruments once you get the preliminary bugs out. I've been running an older E3 for the past few years, but that doesn't make me an expert on them. Your vacuum problems sound a little funny - have you checked that both diff pumps are hot to the touch? If one is cold (the one on the right hand side of the column, viewed from the front, that could explain part of the problem. If it is, try the reset button on its "front" side, and check cooling water flow to make sure it's adequate. Somebody good with electrical stuff (I'm not) can probably determine whether or not your diff pump element is still good. We once had V1 stick a bit, obstructing the beam, but were able to free it up by reversing the compressed air flow momentarily. You can actually watch it open and close if you remove the sheet-metal shroud on the left (the one with the black cylindrical receptacle for the gun for those times when you've pulled the gun - I know, I thought it was a cup holder at first, too, but believe me, it isn't...) The E3 is still supported by FEI, though, and I've always found their guys pretty good about being able to fix things over the phone, even without a service contract. You can get their phone numbers, etc from the site www.feicompany.com. The guys in the Peabody, Mass. office should be able to help you through this one - that's where your E3 was born. Hope this helps.....
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, June 25, 2001 7:11 PM
Julian asked if imaging plates are “as good as negatives”. There are advantages and disadvantages to all imaging systems. In some instances CCD cameras are preferable to film, in other instances film is superior (especially if wide field of view is a factor). What is best to use largely depends on the details of your experiment. I thought I would provide some information on imaging plate systems, and indicate one strong advantage the technology has over other recording media/systems.
Image plate systems use the "photostimulated luminescence" (PSL) phenomenon. Regions in PSL materials that have been previously exposed to a short wave length irradiation, will luminesce when stimulated by a second longer wavelength irradiation. The stored energy is stable until released by the second irradiation. Imaging plates are made using PSL films of barium fluorobromide (= 5 µm grain size), containing a trace amount of bivalent europium as a luminescence center, which are uniformly coated on a polyester support. Imaging plates replace conventional photoemulsion film in the transmission electron microscope and are exposed the same as normal film. To read the recorded image, the imaging plate is latter scanned by a red diode laser and the luminescence is collected through a light collection guide to the photo-multiplier tube (PMT). The analog current produced by the PMT is converted to a 8 to 16 bit depth digital signal. The pixel density can be read at between 50 to 400 pixels/cm. Imaging plates can be fully erased for reuse, but have a finite lifetime.
One main advantage of PSL films in their ultrahigh sensitivity (e.g. 2 x 10-14 - 2 x 10-9 C-cm-2 at 100 kV) to electron irradiation making them valuable for biological work (these are values quoted by a manufacturer Fuji). For example, film emulsions require 0.1 to 1 C-cm-2 to record an optical density S = 1 at 100 k magnification (see Reimer, 1975). Slow scan CCD cameras offer better sensitivity than film, however are inadequate for imaging certain biological specimens. Furthermore, CCD cameras have limited fields of view as compared to film. As an example, Sugi (1997) and coworkers have used a Fuji imaging plate system to record micrographs of myosin head movement induced by ATP hydrolysis in an environmental cell TEM. I avoided using the term “living myosin” since a biomolecule is not strictly alive, but it was biologically active. Electron doses low enough not to damage the delicate biomolecule could be used to record images on an imaging plate.
Reimer, L. (1975) in Physical Aspects of Electron Microscopy and Microbeam Analysis, eds B. M. Siegel, and D. R. Beaman, John Wiley & Sons Publishers, New York, New York, 231-245.
Sugi, H., Akimoto, T., Sutoh, K., Chaen, S., Oishi, N., and Suzuki, S. (1997) Proc. Natl. Acad. Sci. USA 94, 4378-4382.
} } } Does anybody have experience with High Resolution Imaging Plate Scanner } for } } } Electron Microscopy (http://www.ditabis.de). Instead of conventional } } } EM-films, flexible photostimulable phosphor screens of the same size, the } } } so-called Imaging Plates, are loaded into the TEM magazine. Are they as } } } good as negatives? } } } } } } Thanks, } } } Julian } } } } } } _______________________________ } } } Julián Martínez Fernández } } } Senior Research Associate } } } MS 106-5 Ceramic Branch } } } NASA Glenn Research Center } } } Cleveland, OH 44135 } } } Phone: 1-216-433-5512 } } } Fax: 1-216-433-5544 -- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Thanks so much Charles. I just wonder why I did not know that such grids are available. I use JEOL 200CX TEM to look at nanoparticles. What is the thinnest window grid do you recommend?
Regards,
Thearith
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
This has become quite a generic problem for many who are working with nano size particles, and many have found the "solution" to be the use of silicon nitride membrane window grids that have been offered by SPI Supplies for several years. Information about these grids can be found on URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
The silicon nitride is completely structureless and featureless and seems to provide excellent electron transparency. The other advantage is that they are sufficiently robust to survive temperatures in excess of 1000° C. And there is no carbon grain to be confused with the nanoparticles of interest.
Disclaimer: SPI Supplies offers silicon nitride membrane window grids for this type of application.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We are in the process of purchasing an EDS system for our JEOL 100 CX II TEM microscope. We would like to have the EDS system installed at a high angle but we need a "HAXI" kit in order for the EDS system to be installed correctly.
Does anyone have a "HAXI" kit for purchase?
Thank you,
Lou Bustillos AMA Analytical Services, Inc. lbustillos-at-amalab.com
The Image plate system is awesome for materials applications as well.... !!
It is very easy to obtain diffraction patterns that include varying intensities (e.g., a spot pattern mixed with Kikuchi lines) with just one exposure on a single plate. The plates also show the slightest deviations in contrast which are not discernable using film, a CCD camera, or the TEM viewing screen. For example, differences in contrast in amorphous silicon oxide due to the multiple processing steps in integrated circuits may be easily observed with the plates. The advantage in the plates is evident in the low to intermediate magnification ranges where large variations in image contrast can be manipulated off-line. Each IP file is quite large (25MB) and therefore, contains plenty of information which can be manipulated before reducing its size that is suitable for publication purposes. The disadvantages to the IPs are the cost (although I believe the cost is dropping) and the extra processing time needed (~ 80 minutes for a batch of 32 plates).
The CCD camera is still very useful in the high resolution imaging regime where the intensity is rather uniform throughout the field of view and one can instantly observe the quality of the image. We will be evaluating high resolution CCD images with high resolution images obtained with the IPs in the near future. We are not sure what effect (if any) vibrations from the film mechanism will have on the high resolution images.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
The messages from Professor Monson in reply to a question about long-term tissue storage makes me wonder what has happened to this listserver?
His message was highly entertaining and even if the poster of the original message may have felt "picked on", there was no reason to expect him to post an apology, no matter how informative it was.
Far too many posters use this listserver as a way of finding a quick fix to simple solutions to technical problems that can be easily researched in text books.
Often these messages contain little background information on the technical problem under investigation. Even so, many of these questions get full attention and detailed replies, even if the replies are not posted on the main list for public view.
What has happened to the free and open discussion of issues and protocols that we used to have? Are people so afraid of being accused of being incorrect that they do not post their replies openly? Or maybe people are afraid of being accused of being impolite.
I look forward to controversy and hope that my comments are read carefully and replied to with vigor. Only in this way will I be able to learn new thing, and this is the main reason I still subscribe to this listserver.
If I am factually wrong about something, tell me. It is better to find this out on the listserver than in a more professional (or cruel) environment.
As for discussion subjects: why was there no comment about the unadvisable suggestion made about storing samples in a frozen state? Would anyone working with EM really choose to store their samples at -20 degrees before processing for TEM?
Where is all the discussion about extraction that can occur when storing samples in alcohols or buffers, or on the contamination that I know grows in everyones old cacodylate buffers?
Do we not comment because we know it has all been published in textbooks?
Dear Profesor Monson, do not feel bad about improving our general knowledge and our professional etiquette. I for one support your views and find your comments useful and entertaining. Please keep up your submissions.
Best regards,
Paul Webster
Paul Webster, Ph.D. Scientist II and Director Ahmanson Center for Advanced EM & Imaging House Ear Institute 2100 West 3rd Street Los Angeles CA 90057
Scotty Cornelius wrote: ============================================ In a far corner of my mind, I vaguely remember a set of rare earth glass standards that had been developed and were being offered for sale a few years ago, possibly out of the UK. Does anyone know of such a set, or was it all a pipe dream? I'm not referring to the Drake and Weill standards from the early '70s. ================================================ Could you have in mind our (e.g. SPI Supplies) "15 Rare Earth 'Five Phosphates' Standards" for Microanalysis? They can be found on URL http://www.2spi.com/catalog/standards/reepo/reepo.html
At one point, we had inadvertently listed these as "glasses" when in fact they are crystals. So we might have been ourselves the cause of some confusion.
Chuck
Disclaimer: SPI Supplies offers a range of standards for microanalysis including this unique rare earth "five phosphate" mount.
===============================================Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ===============================================
We have just been testing the Ditabis IP scanner and were impressed.
It doesnt quite replace film or I think a good small-pixel below-screen CCD camera for resolution, but is much better than a whole field above screen CCD camera for resolution, and for dynamic range is in a class by itself.
We tested with diffraction patterns showing bright and dim diffuse features, convergent beam patterns, wedge-profile ion beam thinned sections, and normal and unstained biological material.
For low dose work, 1-2 seconds exposure at beam intensities barely visible to the eye was no problem. The resolution problem may be avoidable in practice because images may be taken at higher magnifications and lower beam intensities than normal for film - that is just a question of getting used to matching the imaging conditions to a different medium.
PLate handling is easy. The plates are loaded into the TEM magazine in the light. AFter electron exposure they are kept dark, but loaded into the scanner under light as dim as convenient - we used the light from the monitor screen.
Storing the plates for up to five days made only a very small difference to the intensity histogram - we didnt test beyond that. Plates can also be rescanned with very little loss.
The only real inconvienience we found was the impossibility of recording plate number, mag etc on the IP plate, as we are used to on the negative. If we had our own system we would mark the plates permanently in some way that showed on the readout.
IP plates are complementary to CCD cameras in some respects. You dont get the advantages of immediate viewing, but more dynamic range, more pixels, and of course one scanner can take plates from any number of TEMs. Finally, the price was the sticking point for us at this time. But if a cooperative lab somewhere else on this continent gets one we would likely buy a set of plates and work through the postal system until the price drops or somebody comes up with a "must have" application!
regards Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
} } } Julian Martinez-Fernandez {Julian.Martinez-Fernandez-at-grc.nasa.gov} 06/26/01 05:11AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anybody have experience with High Resolution Imaging Plate Scanner for Electron Microscopy (http://www.ditabis.de). Instead of conventional EM-films, flexible photostimulable phosphor screens of the same size, the so-called Imaging Plates, are loaded into the TEM magazine. Are they as good as negatives?
Thanks, Julian
_______________________________ Julián Martínez Fernández Senior Research Associate MS 106-5 Ceramic Branch NASA Glenn Research Center Cleveland, OH 44135 Phone: 1-216-433-5512 Fax: 1-216-433-5544
I have downloaded from Lumijet4s home page information about the Preservation Monochrome Inks they commercialize for jetink printers, a sort of special ink for good image tonality and permanence. Has any one had any experience with that product? or any other like that?
Regular black ink cartridge would be enough por good quality greyscale printing???
Thanks a lot,
Silvia Montoro Centro Regional de Investigacisn y Desarrollo de Santa Fe (CERIDE) G|emes 3450 3000 Santa Fe
We also have an E3 and have had our share of vacuum problems. Last year the vacuum had gotten to the point that we needed to do something. Another employee and I took all the valves off and removed the diffusion pumps. We cleaned up the pumps and added oil. Then we replaced all the o-rings in the couplings and valves and diff pumps. (basically they were all flat) After that the vacuum was as good as when we first got it and pumping down was much faster. This is something that probably should be done every 10 years whether it needs it or not! We also had some mysterious water loss from the chiller for over a year that finally appeared as a major water leak at the diff pump tubing. We cut off the brittle parts and reconnected. Cooling was better and vacuum too. Of course you should be sure all your vacuum pumps are up to the job, particularly the large pump used for the column. The vacuum hose that goes between the 2 diff pumps should have a metal cover on it to protect it from the heat. It does become brittle and should be checked for leaks. We replaced ours as well. As others have mentioned, the E3 is still supported and although we also do not have a service contract, the engineers at FEI versed in the E3 are VERY helpful, patient and willing to troubleshoot a problem over the phone! Good luck! Regards, Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
If anyone has a Hitachi S-570 with LaB6 that they are getting rid of, we would be interested in any left-over LaB6 crystals mounted in the Wehnelts. Thanks!
Probably today's humor, but hey, it's worth a try.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Paul, While you are correct in stating that the answers to many of the questions asked on this listserver are easily found in a number of available texts, many of the libraries at smaller colleges and universities do not have the funds to maintain an extensive section dedicated to electron microscopy or even microscopy in general. Therefore, those books could be days away via inter-library loan. I always looked at this server as a gathering of some of the great minds in microscopy and a "mother lode" of information based purely on experience, if nothing else. Being a one man show at a small university, I am constantly bombarded with questions about various techniques from investigators throughout the sciences, and have utilized this source many time for answers to those inquiries. There are no stupid questions, only ignorant people asking those questions; and intelligent, informed and timely answers will make those folks somewhat less ignorant. I am now getting down from the soapbox.
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
Our Philips CM-10 transmission electron microscope is now for sale by Michigan State University. For more details, please check the MSU surplus web site:
If you have any other questions, please contact me at: Ph: 517-353-4525. Fax: 517-4325174, Email: fanx-at-msu.edu, We are available for instrument demonstration and examination at any time.
Yours Sincerely, Xudong Fan, Ph.D. Center for Advanced Microscopy Michigan State University East Lansing, MI, 48824
Dear Rachel, Let me add. 1. On storage in buffer: NOT recommended by all my experience and all who have ever advised me. 2. On storage in fixative: NOT recommended as above, always because of unknown variables. 3. On fluorescence microscopy after HCHO: HCHO (dry) has been used to induce fluorescence in so-called biogenic amines (incl., decarboxylated amino acids). One never knows what else it may do during long residence. Ask yourself: Do you want fluorescence of fixed wood or wood? 4. One cannot fix with paraformaldehyde (not your fault). It is a polymer of HCHO and must be depolymerized to yield full methanalic (arch.) power! This (depolymerization) can occur when paraformaldehyde is suspended in a neutral or slightly basic buffer and permitted to stand for a while or heated briefly to 50-60degreesC.
The real identity of paraformaldehyde was the only thing I remember from my failed first course in freight-train organic chemistry [A carload of this and a carload of that yields half a carload of something else!]. A second foray into mechanistic organic went much better.
Summary: Old-timers stored materials to save information. Young folks can't afford the luxury if they want to collect data.
Moral: If you can have some tea while he vents, you might survive to get some solid advice.
Final Regards,
Fred Monson,
} ---------- } From: Rachel Spicer } Sent: Tuesday, June 26, 2001 11:25 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: thanks re: long term storage } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks to all who responded to my post re: long term storage of plant } material in fixatives. The responses were as varied as the recommendations } in the literature, but I find there is no substitute for hearing from } those } with first hand experience. If anyone is interested in the specifics I'd } be happy to forward a collection of relevant responses. And finally, } apologies for my misspelling of "Karnovsky". Thanks again. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
A list of independent EM service providers can be found at "www.jcnabity.com/service.htm".
Note that their "service areas" are not necessarily limited to the "location" that is listed.
If anyone has additional information that can be included on this list, please let me know. Any suggestions for independent EM service providers that are based overseas will also be appreciated.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
In a message dated 6/25/2001 2:59:20 PM Mountain Daylight Time, verlaj-at-medicine.ufl.edu writes:
} Beverly Giammara wrote asking for comments about EM service providers, } other than the manufacturers, that cover the Midwest. I would be very } appreciative if anyone can offer similar information about EM Service } providers that cover Florida. } } Sincerely, } } Jill } } Jill Verlander Reed, D.V.M. } Associate Scientist } Director, College of Medicine Electron Microscopy Core Facility } University of Florida } P.O. Box 100215 } Gainesville, FL 32610 } verlaj-at-medicine.ufl.edu } Phone: (352) 846-0820 } Fax: (352) 846-3299 }
Hello everyone: I have a student who wants to do some immunolocalization of a protein in plant tissue. I have been combing my files, but haven't managed to find a decent protocol that she could work from... (and can't seem to get myself to the library for a more comprehensive search). Can anyone supply us with a "generic" protocol for immunofluorescence in plants?
A customer of mine sent a part that has started showing a failure only recently, although he has manufactured the part for years. The part is a heating element. It consists of a thick SS rod which is the terminal. It has a diameter of about 1 cm. It is joined to an element that is about 2 mm in diameter. The joint is made by 1st drilling the thick stock and then inserting the wire into the hole. Then a notch is made in the thick stock until the thin element is exposed. Then the notched surface is welded without the introduction of any new material. I believe it is a "TIG" weld. A standardless analysis of the terminal shows Fe (82%), Cr(13%), Al(3%), Si(1.6%), and Mn (.3%). He has tried to match the two materials as closely as possible.
The joint is heated repeatedly, as it is used as an element in a glass furnace. The failure starts with a bulk flow of material out the top of the hole, which is about 1.5-2.0 cm from the welded joint/notch, and over the sides of the terminal. I have analyzed this material and it contains principally Fe and O. The area I analyzed was polished so the internal composition of the flowed material contains a great deal of oxygen. No Cr is detected and very little Al.
I feel that this material flow degrades the joint causing the various electrical failures that have been observed.
My questions are:
Is this a typical failure mode with a Fe/Cr SS weld experiencing many heat cycles?
What is the mechanism that results with the crystalline appearance of the iron oxide, leaving the other elements behind?
This failures appears to have started when a new aqueous lubricant was introduced for the initial hole drilling of the terminal. Could this play a role in the failure?
What measures might be taken to reduce the frequency of this failure mode? Joint design? Material compatibility?
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
I would like to gather some information from the micromanipulator community. Specifically, this tool will be used for lamella extraction after the FIB work is done. We will be in the market to purchase one in the upcoming months so I'm starting my homework early. Ideally, this will be a dedicated station with an anti vibration table along with the optical microscope. Some questions are: 1) Motorized vs manual 2) Objective lens working distance 3) Probe resolution 4) Sensitivity (microns per degree)
Any comments or suggestions are welcomed!
Thank you,
Jon Hiller
================================================================== Jon M. Hiller Argonne National Laboratory Materials Science Division Electron Microscopy Center Tel: 630-252-9558 Fax: 630-252-4798 Email: hiller-at-anl.gov ==================================================================
There is a wonderful protocol for immunofluorescence in a manual which I use for most of my Arabidopsis work.
Methods in Plant Molecular Biology A laboratory course manual Maliga, Klessig, Cashmore, Gruissem, Varner Cold Spring Harbor Lab Press
Good luck,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Wed, 27 Jun 2001, Shea Miller wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone: } I have a student who wants to do some immunolocalization of a protein in plant tissue. I have been combing my files, but haven't managed to find a decent protocol that she could work from... (and can't seem to get myself to the library for a more comprehensive search). Can anyone supply us with a "generic" protocol for immunofluorescence in plants? } } Thanks so much in advance } shea } } } } }
What is the composition of the heating element? The S.Steel terminal is not a common (to me) stainless, given the composition you obtained. I would have to check references to see what it may match. I work mostly with austenitic stainless and nickel super alloys when it comes to "SS".
I suspect the welling up of oxide from the joint/hole is a symptom rather than a cause of the failure. Contact area and/or conductor size reduction will (all else being the same) cause a hot spot. What is the shape of the FeOxide crystals? Sounds like you may have magnetite, a stochiometry often formed under oxygen starved (but still oxidizing) environment.
I would question the customer carefully about process conditions which result in failure. What is the composition of the lube? If residue remains, it could surely play a role in the corrosion. Are there any differences in glass chemistry or temperatures compared to the non-failed parts? What environment is "seen" by the joint? -} In the Glass, near it? Glass chemistry? If near it, what is the atmosphere? Temperatures? Heating power/time curve the same (i.e. any localized I2R heating of joint)? Getting accurate and complete information from customers is sometimes the most difficult part of failure analysis. ...Especially if it is second hand.
Too much weld heat can "sensitize" some alloys (changes carbon/carbides), reducing resistance to corrosives. The failure mode, however, is most often intergranular cracking and (again) I am not familiar with the characteristics of your composition.
So many questions, so few answers, sorry!
Woody
} -----Original Message----- } From: Smartech [mailto:smartech-at-optonline.net] } Sent: Wednesday, June 27, 2001 1:33 PM } To: To all on the list } Subject: Metallurgy, Weld Failure Analysis, SEM Analysis } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } A customer of mine sent a part that has started showing a failure only } recently, although he has manufactured the part for years. } The part is a } heating element. It consists of a thick SS rod which is the } terminal. It } has a diameter of about 1 cm. It is joined to an element } that is about 2 mm } in diameter. The joint is made by 1st drilling the thick } stock and then } inserting the wire into the hole. Then a notch is made in } the thick stock } until the thin element is exposed. Then the notched surface is welded } without the introduction of any new material. I believe it } is a "TIG" weld. } A standardless analysis of the terminal shows Fe (82%), } Cr(13%), Al(3%), } Si(1.6%), and Mn (.3%). He has tried to match the two } materials as closely } as possible. } } The joint is heated repeatedly, as it is used as an element in a glass } furnace. The failure starts with a bulk flow of material out } the top of the } hole, which is about 1.5-2.0 cm from the welded joint/notch, } and over the } sides of the terminal. I have analyzed this material and it contains } principally Fe and O. The area I analyzed was polished so } the internal } composition of the flowed material contains a great deal of } oxygen. No Cr } is detected and very little Al. } } I feel that this material flow degrades the joint causing the various } electrical failures that have been observed. } } My questions are: } } Is this a typical failure mode with a Fe/Cr SS weld } experiencing many heat } cycles? } } What is the mechanism that results with the crystalline } appearance of the } iron oxide, leaving the other elements behind? } } This failures appears to have started when a new aqueous lubricant was } introduced for the initial hole drilling of the terminal. } Could this play a } role in the failure? } } What measures might be taken to reduce the frequency of this } failure mode? } Joint design? Material compatibility? } } } Thanks in advance! } } Ric } } SMARTech } 860-491-3299 } www.semguy.com } 19 Cornwall Drive } Goshen CT 06756 } }
I'm in search of something that will remove immersion oil from slides of
blood smears (no coverslips) without damaging the specimens. A friend told that such a chemical exists, but could not remember the name. It supposedly smells like oranges, and xylene was once used for the same purpose. If you have any ideas as to the name of this mystery chemical,
Does anyone know the best method for embedding 0.45um nitrocellose filter for TEM thin sectioning? We have been filtering vesicle preparations onto these filters and wish to examine them by TEM analysis for immunolabelling. Our 2 previous attempts with Epon mixtures and minimal intermediate solvent changes have lead to poorly polymerized blocks and dissolving membranes. Currently we are trying LR White as a substitute. Any help would be great. Thank you. Ingrid Niesman SRA III Farquhar/Palade Lab UCSD
Dear all, Is the DTSA (Desk Top Spectrum Analyzer) package available for PC or only for Mac? I looked on the NIST web site but couldn't find information (and their internal search engine was broken).
Thanks for any leads you can give.
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I have no idea if it works for your purpose, but the xylene substitute smelling of oranges is called Histoclear.
Dave
On Wed, 27 Jun 2001 17:42:41 -0500 Christine Edly {cedly-at-ou.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings! } } I'm in search of something that will remove immersion oil from slides of } } blood smears (no coverslips) without damaging the specimens. A friend } told that such a chemical exists, but could not remember the name. It } supposedly smells like oranges, and xylene was once used for the same } purpose. If you have any ideas as to the name of this mystery chemical, } } I'd sure appreciate it. } } Regards, } Christine Edly } University of Oklahoma }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have used the 'orangey' smelling solvent under the brandname of Histoclear. It is supposedly extracted large scale from citrus peel and is the composed of the 'zest' that comes out as a spray when we peel oranges.
I couldn't comment on its use for removing immersion oil as you describe. I have only used it as a xylene substitute for tissue processing tissues and also for clearing stained sections prior to mounting.
It is said to be safer than xylene but some of the people I worked with at the time complained of headaches so we stopped using it despite never proving that it was the cause of the headaches.
Hope this helps!
At 17:42 2001-06-27 -0500, Christine Edly wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
The point about this server is that it is used by a very heterogeneous public from which a non-exclusive list is the following:
students, professors, scientists/ researchers technical staffs suppliers producers specialists non-specialists etc. etc. etc.
and all these from so many different backgrounds with so many different problems and even from so many different countries... That if we want to teach all of them to behave as we expect before they post a query, the server could go to early retirement.
I guess the philosophical attitude that one should have is to really try and understand both sides of the discussion.
1- on one hand the trust that professor Monson is demanding is not there because our users are a small sample of our society. How many times have we heard that people have had their ideas picked by others and their publications stolen by others and .... in the very scientific community that most of us hoped to be the purest part of the society. Still the scientists are generally speaking very sincere people. So if someone is not having trust to others, it's the consequence of what they have observed in their environment.
2- I do not know if Rachel is a student (may be not), and if she really was not trusting or just did not think to write the reason and so on, so we do not discuss the case on a specific way. But talking about going to the library, there are labs where the students have really not access to the books and there are places where superiors do not give the support the students and other researchers' need, or simply there are not enough books available.
So there are always reasons for the way people behave and the more our society goes towards a very materialistic and "win whatever the situation" attitude, the more the arguments of professor Manson will be applicable because the more people will behave in the way he is not happy about, and I understand and apreciate his point.
All that apart, it is better to listen to people,as often things that are not to be done are not published and are only "known". Of course provided that answers are given generously, as arrogance for giving is so destructive, even for donator. I therefore agree with the fact that we should just be able to ask the question and someone will react to give us an answer.
I think that professor Monson has evoked a point that is of great importance and that is the human aspect of all we do, as "human" is what we are hoping to be at a first place. That needs sensitivity... and continuous effort.
Regards,
Sousan
Paul Webster wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Colleagues, } } The messages from Professor Monson in reply to a question about long-term } tissue storage makes me wonder what has happened to this listserver? } } His message was highly entertaining and even if the poster of the original } message may have felt "picked on", there was no reason to expect him to post } an apology, no matter how informative it was. } } Far too many posters use this listserver as a way of finding a quick fix to } simple solutions to technical problems that can be easily researched in text } books. } } Often these messages contain little background information on the technical } problem under investigation. Even so, many of these questions get full } attention and detailed replies, even if the replies are not posted on the } main list for public view. } } What has happened to the free and open discussion of issues and protocols } that we used to have? Are people so afraid of being accused of being } incorrect that they do not post their replies openly? Or maybe people are } afraid of being accused of being impolite. } } I look forward to controversy and hope that my comments are read carefully } and replied to with vigor. Only in this way will I be able to learn new } thing, and this is the main reason I still subscribe to this listserver. } } If I am factually wrong about something, tell me. It is better to find this } out on the listserver than in a more professional (or cruel) environment. } } As for discussion subjects: why was there no comment about the unadvisable } suggestion made about storing samples in a frozen state? Would anyone } working with EM really choose to store their samples at -20 degrees before } processing for TEM? } } Where is all the discussion about extraction that can occur when storing } samples in alcohols or buffers, or on the contamination that I know grows in } everyones old cacodylate buffers? } } Do we not comment because we know it has all been published in textbooks? } } Dear Profesor Monson, do not feel bad about improving our general knowledge } and our professional etiquette. I for one support your views and find your } comments useful and entertaining. Please keep up your submissions. } } Best regards, } } Paul Webster } } Paul Webster, Ph.D. } Scientist II and Director } Ahmanson Center for Advanced EM & Imaging } House Ear Institute } 2100 West 3rd Street } Los Angeles } CA 90057 } } pwebster-at-hei.org } p: 213 273 8026 } f: 213 413 6739 } http://www.hei.org/research/depts/aemi/aemi.htm
The micromanipulator we use is a manual hydraulic system (Narishige MNN-1) which is suitable for the plucking process. I do not know how its performance compares of to other makes.
We use a microscope with *50 and * 5, 18 mm working distance lenses.
A couple of general comments about the lift-out process which might be useful.
We use both pulled glass needles and electro-polished metal needles. The yield of plucking using these two types of needles is comparable. An advantage of the metal needles is that they do not shatter if accidentally knocked against the sides of the FIB cuts. An advantage of the glass needles is that if a cross-section has moved up the needle's shank, away from the tip, (which sometimes occurs) then it is possible to see it and therefore, not waste time searching for it in the area surrounding the FIB cuts. (In addition to riding up the needle's shank sometimes a cross-section can also 'flick ' out away from the cuts while it is being plucked).
For cross-sections that have moved up the shank it is possible to re-position them at the needle's tip using a second micro-manipulator. However, it is also possible to get these cross-sections off the needle and onto the support TEM grid by gently tapping the micro-manipulator system while the needle is positioned over the grid.
Instead of using the Formvar or carbon support grids we use copper grids with 7 um sized holes. We have found that only very rarely does a cross-section fall off these. The advantage of these is that there is no support membrane. What can happen is that while a cross-section is being placed onto the support membrane the support membrane can be torn by the tip of the needle and it can then wrap itself around the cross-section.
Also, because of the diversity of use of FIB systems (e.g. for modification or fabrication of optoelectronic devices and micromachining) a FIB discussion list has been set up. The subscribing web page is located at:
http://users.ox.ac.uk/~rml/
Any information, for example details of relevant FIB pages, can be added to the web page if emailed to me.
Regards
Richard
-------------------------------------------------------------- Richard M Langford
Department of Materials, University of Oxford Parks Road, Oxford, OX1 3PH, UK
I agree wholeheartedly with Paul in his comments regarding the mails from Frederick Monson - no need for apology, interesting and informative replies.
Higher education seems to be locked in an apparently paradoxical situation with students being expected to 'take responsibility for their own learning', the use of Student Centred & Problem Based Learning etc. The paradoxical part is that they do not have the appropriate intellectual tools to be able to carry out such assignments. Frustration with their lot then spills out over tutors who are expected to take responsibility when things go wrong (but mostly no feedback when things work).
Ask me a question and I will give you the most appropriate answer I can. The answer may be wrong of course, possibly because I have an incorrect understanding of the concepts involved, possibly because I was asked the wrong question. If I respond to a question with a question of my own it is because I am trying to seek clarification of what EXACTLY is being asked. That, as I see it, is how we work in science.
Paul says - "If I am factually wrong about something, tell me. It is better to find this out on the listserver than in a more professional (or cruel) environment."
I agree absolutely here too - it is easier to fight tooth and claw here and come to some agreement or compromise than if we are addressing 500 people at a conference and one of them asks a simple question to which we have no answer.
My appeal to those posing questions is to give as much background information as possible and to respond clearly to questions on clarification.
My appeal to those answering is to identify clearly what is hearsay "in my experience" "I got that straight from a guy in the pub last night". Identify also the context of the answer/solution to the question/problem and refer to published data whenever possible.
I look forward to a lively, ongoing debate (even though I am going for my summer break tomorrow!).
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
} My boss just noticed in the 1985 edition of Cellular Pathology Technique, } Culling, Allison and Barr eds., that food oil derivatives, such as } histoclear, will cause "haematoxylin" staining to rapidly fade. Is this } true? I have been having problems with hematoxylin fading for quite some } time and I use histoclear in my paramount all the time. 'Tho I began (8 } years ago) by using xylene, and I think those faded just as much. } } If histoclear does cause the stain to fade, are there any other } substitutes for xylene that would work?
R. D. Lillie made extensive studies of mounting and fading in the 1940s and 1950s (reported in Ch. 5 of his "Histopathologic Technic" book, with references to papers in Stain Technol).
Fading is typically due to either acidity or chemical reduction. Canada balsam is acidic (abietic acid is a major component) but this doesn't matter - the acid can't ionize in the absence of water. Alum-haematoxylin stains keep for many years in Canada balsam though an eosin counterstain may fade a little over the course of 2 years. (This must be from reducing agents in the medium.) Easily reduced colours fade in Balsam (e.g. Prussian blue, acid fuchsine) and are more stable in synthetic media. Easily oxidized colours (notably cobalt sulphide) are well preserved in Canada balsam but fade in a few months in most synthetic resins. (You can revive CoS by removing the coverslip and exposing to ammonium sulphide again.)
Limonene (I think this is what histoclear is) is a reducing agent - two non-conjugated double bonds in the molecule - so its presence in a mounting medium is undesirable. Some is bound to persist, just as some xylene persists when the mounting medium dries. The boiling point of limonene (176C) is a bit higher than xylene (140C), so it might be a bit more persistent.
If your eosin faded equally after xylene or limonene the mounting medium is the probable culprit. Try a wholly synthetic (polystyrene or methacrylate) medium rather than a natural or semi-synthetic one. Synthetic media are the best bet for pretty well everything except cobalt sulphide (from ATPase histochemistry etc). The blue component (alum-haematein) of an H & E stain is very resilient and should not be affected by clearing agents and mounting media. It keeps for decades (perhaps for ever). If yours is fading, something acidic and probably also some alcohol must be passing through into the mounted preparations. This is surely unlikely to occur on a regular basis!
Hope this helps. I'm sure I haven't thought of everything, and have no personal experience with histoclear/limonene, but you should be getting lots of other suggestions too.
John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1
Hi
I have used the 'orangey' smelling solvent under the brandname of Histoclear. It is supposedly extracted large scale from citrus peel and is the composed of the 'zest' that comes out as a spray when we peel oranges.
I couldn't comment on its use for removing immersion oil as you describe. I have only used it as a xylene substitute for tissue processing tissues and also for clearing stained sections prior to mounting.
It is said to be safer than xylene but some of the people I worked with at the time complained of headaches so we stopped using it despite never proving that it was the cause of the headaches.
Hope this helps!
At 17:42 2001-06-27 -0500, Christine Edly wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
It would be helpful to know what brand the knives are. Most knife manufacturers will take back their own knives for sharpening at the factory. Just find the local distributor for the brand of knife you have and they will arrange things for you. Energy Beam Sciences is a U.S. Company. We recommend and deal primarily with Diatome but we can also help with DDK.
Good luck, Mike
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: Ping Li [mailto:pli-at-is.dal.ca] Sent: Wednesday, June 27, 2001 4:10 PM To: Microscopy list
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
Just wanted to thank everyone who contributed to the discussion and suggested solutions for the problems we were having. It is very much appreciated - we have so many things to try and have learned a lot in the process.
My colleague, who shares the printer with me, was amazed, both with the range of answers and the rapidity of the responses - the perfect combination when you're having trouble in the lab. That's why I keep coming back to the group for answers and have stayed subscribed for so many years - there's no place like it!
Thanks again.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
When we do this sort of thing, we generally use Nuclepore filters, but I think our trick should work for other filters as well. We keep the material on the filters through fixation and change solutions with gentle vacuum. Then we overlay the specimen with a thin layer of low gelling point agarose, pull a gentle vacuum and then chill the filter to solidify the agarose. The entire filter can then be submerged in the next fluid, and the filter is peeled away from the agarose. If all goes well the specimen should be trapped in the agarose and the filter material is no longer a problem. Warning: do not fix agarose in glutaraldehyde. It does not embed well.
At 09:20 PM 6/27/2001 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
This is a reminder concerning upcoming deadlines for the Microscopy & Microanalysis 2001 meeting from August 5-9 in Long Beach, CA.
1) The deadline for hotel reservations at the meeting rate is this Friday, June 29. The Westin still has some double rooms available for the entire meeting period, but the other meeting hotels are booked. You can reserve a room online by going to www.msa.microscopy.com and following the links to M&M 2001 electronic forms.
2) The deadline for early meeting registration is Friday, July 6. Save some money and register early! Electronic registration forms are also available at the above site. I would also encourage those of you that are interested to register for the Pre-meeting congress (only $10 more with full meeting registration) and workshops ($120-$180) that are available.
3) The deadline for submitting a late breaking poster is Sunday, July 15th. If you would like to submit your work for this session contact Bob Price by email at Price-at-med.sc.edu.
With nearly 700 presentations scheduled, a sold-out exhibit floor and an excellent venue in Long Beach, California, M&M 2001 promises to be an excellent meeting. On behalf of the MSA and MAS Councils, and the Local Arrangements and Program Committees, I hope to see all of you there.
Christine, if it smells like oranges and is used instead of xylene it must be Histoclear. I have mine from Electron Microscopy Sciences, called Xylene substitute, Cat #23410-01. I have never used it for immersion oil removal though (sounds like a good idea), I just use it to clean microtomes of paraffin residue.
Sarka Lhotak Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
} Greetings! } } I'm in search of something that will remove immersion oil from slides of } } blood smears (no coverslips) without damaging the specimens. A friend } told that such a chemical exists, but could not remember the name. It } supposedly smells like oranges, and xylene was once used for the same } purpose. If you have any ideas as to the name of this mystery chemical, } } I'd sure appreciate it. } } Regards, } Christine Edly } University of Oklahoma } }
We are in the market for a freeze-fracture system. Currently there are two models under our consideration: Bal-tec Freeze-Fracture Unit BAF060 and RMC/JEOL 9010. We would appreciate any comments from users of these two systems regarding to their applications, performance, stability, and other features.
Ingrid: It has been a long time since I did this, but my notes show that we have very good embedding using Epon, _but_ I have omitted propylene oxide from my processing schedule (did that more due to contact allergies developed over years of carelessness). Epon (and the various replacements are miscible with EtOH--just make sure that you use freshly opened bottles. (Acetone is another dehydrant that will tend to dissolve the filters, BTW.) Dehydration steps can be shortened (through graded EtOH, including uranyl acetate in 70% if you use a preembedding UA procedure), then go into 2:1, 1:1 and 1:2 EtOH:Epon for infiltration. I also use a minimum of 2 steps in pure before embedding in fresh epoxy. I would guess that you are embedding in the flat molds--best way for filters, IMHO. From personal experience, use of PO and Spurr's are really out for the filters (and my recollection is the same for acetone, as mentioned above). Hope it works out for you.
Roger
On Wed, 27 Jun 2001 21:20:19 -0700, Ingrid R. Niesman wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Does anyone know the best method for embedding 0.45um nitrocellose filter for TEM | thin sectioning? We have been filtering vesicle preparations onto these filters and | wish to examine them by TEM analysis for immunolabelling. Our 2 previous attempts | with Epon mixtures and minimal intermediate solvent changes have lead to poorly | polymerized blocks and dissolving membranes. Currently we are trying LR White as a | substitute. Any help would be great. | Thank you. | Ingrid Niesman | SRA III Farquhar/Palade Lab | UCSD |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I would appreciate any responses about the following situation:
I have epoxy resin embedded plant leaf (pepper) material infected with a plant pathogenic bacteria (Xanthomonas vessicatoria). We plan to begin with cutting 1-2 micron sections for light microscopy. I know that there are stains using for differential staining of bacteria which are used on fresh tissue or paraffin embedded material. Does any know of stains which would work similarly for epoxy resin embedded material?
Thank you,
Maureen Petersen Dept of Plant Pathology University of Florida Gainesville, FL
Fixation? You shouldn't partially dehydrate or partially clear for EPON, but assuming the membrane system is adequately preserved to survive the alcohol and PropOx, then your problem is poor infiltration, due to poor clearing, due to poor dehydration. This does not mean that you must dehydrate in 30min steps. The times can be reduced empirically until you get to the minimum necessary times for dehydration and clearing. Lots of variables. The filter should not be a problem in any case, though I have no practice with isolated vesicles, only with isolated Eprythrozoon coccoides (a mouse zoonose, something like a mycoplasma, likes RBC's, and 'hibernates') and just morphology. You might want to try GMA for TEM (e.g. SPI) though I have no experience with it, or any other hydrophilic embedment. I assume nano-gold, so you must have been into: Hayat, M.A.(1995), Immunogold-Silver Staining, CRC Press, Boca Raton, FL, ISBN: 0-8493-2449-1(LC Card# 95-1831, LC Loc# QR187.I482H39). Chapter 4 by Krenacs and Krenacs on the subject of: "Which embedding medium?" might be of help.
Good hunting,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Ingrid R. Niesman } Sent: Thursday, June 28, 2001 12:20 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: nitrocellulose filters } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone know the best method for embedding 0.45um nitrocellose filter } for TEM } thin sectioning? We have been filtering vesicle preparations onto these } filters and } wish to examine them by TEM analysis for immunolabelling. Our 2 previous } attempts } with Epon mixtures and minimal intermediate solvent changes have lead to } poorly } polymerized blocks and dissolving membranes. Currently we are trying LR } White as a } substitute. Any help would be great. } Thank you. } Ingrid Niesman } SRA III Farquhar/Palade Lab } UCSD } }
This is very interesting, about the orange or citrus-based stuff that is good for removing immersion oil. There is also a range of products sold for de-greasing bicycle chains and derailleurs, which are based on citrus components. I'm no organic chemist myself, but can one of the chemistry-oriented people out there tell me why something citrus-based should be so good at dispersing oils? Call me curious.....and I hope I'm not too off-topic here..
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia B3L 4C8
No Protocol, but this was a quick find on Google "Plant immunohistochemistry"
Plant Cell Physiol. 37, 641-649 (1996): Developmental expression of a 23 kDa jasmonate-induced protein (JIP-23) of barley leaves (Hordeum vulgare cv. Salome) was studied by measuring the time-dependent accumulation of transcript and protein during germination. Tissue-specific expression of JIP-23 was analyzed immunocytochemically and by in situ hybridizations, respectively. During seed germination JIP-23 mRNA was found to accumulate transiently with a maximum at 32 h, whereas the protein was steadily detectable after the onset of expression. The occurrence of new isoforms of JIP-23 during germination in comparison to jasmonate-treated leaves suggests, that the JIP-23 gene family of barley is able to express different subsets of isoforms dependent on the developmental stage. JIP-23 and its transcript were found mainly in the scutellum, the scutellar nodule and in lower parts of the primary leaf of 6 days old seedlings. All these tissues exhibited high levels of endogenous jasmonates. In situ hybridization revealed specific accumulation of JIP-23 mRNA in companion cells of the phloem in the nodule plate of the scutellum. In accordance with that, JIP-23 was detected immunocytochemically in phloem cells of the root as well as of the scutellar nodule and in parenchymatic cells of the scutellum. The cell type-specific occurrence of JIP-23 was restricted to cells, which are known to be highly stressed osmotically by active solute transport. This observation suggests, that the expression of this protein might be a response to osmotic stress during development.
URL: http://www.ipb-halle.de/english/institute/research/hause/introduction.htm
Interesting,
Fred Monson
} ---------- } From: Shea Miller } Sent: Wednesday, June 27, 2001 12:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: immunofluorescence on plants } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone: } I have a student who wants to do some immunolocalization of a protein } in plant tissue. I have been combing my files, but haven't managed to } find a decent protocol that she could work from... (and can't seem to get } myself to the library for a more comprehensive search). Can anyone supply } us with a "generic" protocol for immunofluorescence in plants? } } Thanks so much in advance } shea } } } } }
Question from an old exam given to students who demanded MORE MULTIPLE CHOICE!
Which of the following is NOT involved with human reproduction?
a. transcription b. replication c. translation d. osculation e. none of the above
Answer tomorrow.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Ian MacLaren } Reply To: Ian MacLaren } Sent: Thursday, June 28, 2001 3:56 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: DTSA } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } Is the DTSA (Desk Top Spectrum Analyzer) package available for PC or only } for Mac? I looked on the NIST web site but couldn't find information (and } their internal search engine was broken). } } Thanks for any leads you can give. } } Ian MacLaren } Max Planck Institut f¸r Metallforschung, Seestrae 92, } 70174 Stuttgart, Germany } Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk } Home page: http://members.tripod.co.uk/IanMacLaren/ } } }
Please post informational responses to the server! I've been curious about this, too but too lazy to look into it - the dishwashing cleansers sold in "granola" stores (organic, non-toxic to the environment, etc.) are also citrus-oil based (and at least one has almond oil; not sure if that is just for the odor).
Tamara
On Thu, 28 Jun 2001, Frank Thomas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listers - } } This is very interesting, about the orange or citrus-based stuff that is } good for removing immersion oil. There is also a range of products sold for } de-greasing bicycle chains and derailleurs, which are based on citrus } components. I'm no organic chemist myself, but can one of the } chemistry-oriented people out there tell me why something citrus-based } should be so good at dispersing oils? } Call me curious.....and I hope I'm not too off-topic here.. } } Frank Thomas } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } B3L 4C8 } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Michael Haugh - Director of Operations Resolution - http://www.resolve3d.com 530 Tamal Plaza, Corte Madera, CA 94925 Office:415/945-7360 x13 FAX:415/927-4495 Cell:415/987-9929 Email: mhaugh-at-resolve3d.com
-----Original Message----- } From: Ping Li [mailto:pli-at-is.dal.ca] Sent: Wednesday, June 27, 2001 1:10 PM To: Microscopy list
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
Thanks so much for all the info on protocols for plant immunofluorescence... we tried a trial run today on some immature soybeans embedded in paraffin, and got some positive results following the general methods of Tobias Baskin. You have all given great advice, and we have lots to work with to refine our immunoassay.
thanks so much shea
Dr. S. Shea Miller Agriculture & Agri-food Canada Eastern Cereal and Oilseed Research Centre Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 Email: millers-at-em.agr.ca
Micronetsters, I am interested in this thread about removing immersion oil, though perhaps diverged a bit from the original post. Over the years, we have often put oil on a slide and then wished to look without. In my hands, this is never easy. I have taken chloroform, acetone, and the like to the coverslip, and wiped and wiped, and it takes truly many wipes to get oil well and truly off. Small beadlets of oil remain tenaciously stuck to the glass. If someone does have a simpler way to get oil off a slide, so well off that there is no residue left behind, I'd love to know about it.
We use Micro Star Technologies for sharpening diamond knives. They do a good job and their turnaround time is very good.
Micro Star Technologies 511 FM 3179 Huntsville, TX 77340-2069 Phone 800/533-2509 FAX 409/294-9861 email mistar-at-msn.com
Mike
Michael Haugh - Director of Operations Resolution - http://www.resolve3d.com 530 Tamal Plaza, Corte Madera, CA 94925 Office:415/945-7360 x13 FAX:415/927-4495 Cell:415/987-9929 Email: mhaugh-at-resolve3d.com
-----Original Message----- } From: Mike Haugh Sent: Thursday, June 28, 2001 1:15 PM To: 'Ping Li'; Microscopy list
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
Sparkle! The wonderful purple glass cleaner stuff. Not available in all parts of the US, even - I was given a stash by a student from the midwest. I've heard stories of microscope company service people back east who stockpile the stuff (definitely couldn't find it in New York). I think the guy who gave it to me said it was available in True Value hardware stores; I don't know if they make it.
No financial attachments to Sparkle or True Value, unfortunately.
Tamara
On Thu, 28 Jun 2001, Tobias Baskin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Micronetsters, } I am interested in this thread about removing immersion } oil, though perhaps diverged a bit from the original post. Over the } years, we have often put oil on a slide and then wished to look } without. In my hands, this is never easy. I have taken chloroform, } acetone, and the like to the coverslip, and wiped and wiped, and it } takes truly many wipes to get oil well and truly off. Small beadlets } of oil remain tenaciously stuck to the glass. If someone does have a } simpler way to get oil off a slide, so well off that there is no } residue left behind, I'd love to know about it. } } Thanks, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123 } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
My guess about limonene is that it is non-toxic, biodegradeable, easily detectable by (pleasant) smell, a 'waste' product of the food industry, doesn't need to be mined (environmentally friendly) etc ......
I have seen the bike chain cleaners too and my first reaction was to think Histoclear!
Have a good summer - I'm off to Finland camping on Sunday - Yippeeeeeeee!
At 13:41 2001-06-28 -0300, Frank Thomas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Micronetsters, } I am interested in this thread about removing immersion } oil, though perhaps diverged a bit from the original post. Over the } years, we have often put oil on a slide and then wished to look } without. In my hands, this is never easy. I have taken chloroform, } acetone, and the like to the coverslip, and wiped and wiped, and it } takes truly many wipes to get oil well and truly off. Small beadlets } of oil remain tenaciously stuck to the glass. If someone does have a } simpler way to get oil off a slide, so well off that there is no } residue left behind, I'd love to know about it. } } Thanks, } Tobias }
I have try solvents from alcohol to ammonia and thanks to the advice of the local confocal Leica rep. Frank Lie, I am now using Windex with great success for removing immersion oil from oil objectives and COVERSLIPPED microscope slides.
As for using Histoclear, which I have: it is rather strong smelling and I am uncertain whether it will destain uncoverslipped stained histo. preps. Cheers, Ken
-- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
At 4:28 PM -0500 6/28/01, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Micronetsters, } I am interested in this thread about removing immersion } oil, though perhaps diverged a bit from the original post. Over the } years, we have often put oil on a slide and then wished to look } without. In my hands, this is never easy. I have taken chloroform, } acetone, and the like to the coverslip, and wiped and wiped, and it } takes truly many wipes to get oil well and truly off. Small beadlets } of oil remain tenaciously stuck to the glass. If someone does have a } simpler way to get oil off a slide, so well off that there is no } residue left behind, I'd love to know about it. } } Thanks, } Tobias } -- *********************
For years, we have used a 1:1 dilution of Windex & water. Use the original formula (blue) Windex. this was recommended to use by our Zeiss representatives and works well on slides and lenses.
lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} } Hi, could anyone please give me some information on where I can get my } diamond knives (for semi thin sectioning) sharpened? Any such places in } Canada? I believe the knives were made in Germany. Any information is } highly appreciated. Thank you in advance. } } Ping *****************
I believe that almost all of the companies that manufacture & sell diamond knives (Diatome, DDK, Mictostar) will resharpen knives of all makes. All are the US or have US offices.
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Sparkle glass cleaner is good for cleaning the larger glass surfaces of the microscope. I don't think it would really work well to remove immersion oil, we use acetone for that. Acetone is a low health hazard (fingernail polish remover) and works well for cleaning objectives and eyepieces. Like any cleaner, even Sparkle, you would never soak an optical element. Apply the cleaner to something soft and absorbent and use that to clean the surface. Kimwipes and wooden shafted "Q-tips" work the best. It's true that Sparkle isn't available everywhere. You can order it from their website: http://www.glasscleaner.com/
Dave Burton Optical Specialist University of Washington Scientific Instruments
Just wondering: isn't acetone likely to attack the cements holding the lens elements in place? Or even some plastic components? Apparently you haven't had any problems, but I wonder if anyone else has?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: David Burton [mailto:dburton-at-nwlink.com] Sent: Friday, June 29, 2001 8:40 AM To: Microscopy-at-sparc5.microscopy.com
Sparkle glass cleaner is good for cleaning the larger glass surfaces of the microscope. I don't think it would really work well to remove immersion oil, we use acetone for that. Acetone is a low health hazard (fingernail polish remover) and works well for cleaning objectives and eyepieces. Like any cleaner, even Sparkle, you would never soak an optical element. Apply the cleaner to something soft and absorbent and use that to clean the surface. Kimwipes and wooden shafted "Q-tips" work the best. It's true that Sparkle isn't available everywhere. You can order it from their website: http://www.glasscleaner.com/
Dave Burton Optical Specialist University of Washington Scientific Instruments
Fax support is not built into Outlook 2000 and the newer version of Windows as it was in the older versions. MS does describe the setup on their knowledge base. Basdically, it is still available by going back and installing some add-ons that are not part of the regular Windows or Outlook installation. They are in one of those extra folders that you find on the CDs. I will send you details directly.
At 09:34 AM 6/29/2001 +1000, you wrote:
} Dear colleagues, } } I know that here is some very good experts on computers.. And I cannot } configuration how to do it.. } } I would like to know how to change setting of the fax in Outlook 2000 from } email only to send and receive faxes.. } } Ricardo
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Just a reminder that the hotels are filling up, & the registration rate will be going up next week for TNT 2001 (Segovia, Spain, September 3-7th). There are currently some 250 participants from all over the globe, from the fields of physics, microscopy, FIB, biotech, materials and the semiconductor industry, as well as dedicated Nanotechnologists and a smattering of venture capitalists.
More information is at www.cmp-cientifica.com/tnt2001
Regards
Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
1) Avery 9x12" Self - Adhesive Laminating sheets, 50/ pack. #461253 / 73601 The result is a nice glossy finish, and an image that doesn't fade.
2) archival printer ink. Check the Epson web site for more info about archival inks and printers like the Epson StylusPhoto 2000P. Wilheim research does the most testing for archivalness of photographic media. They may have some pointers on the best ink-paper combos for light stability. http://www.wilheim-research.com
3) enclosing a print in a polyethylene sheet protector (archival grade)
4) printer specific paper -Stay with the papers recommended for use with the new Epson inks
- try Epson's Heavyweight Matte paper It's cheap ($9/50 sheets from a discount sppplier), delivers the Epson long-life guarantee, & it has a robust surface right out of the printer. And beautiful prints.
-There were recently (last 6-8 months) some issues with color shift on some Epson papers. Epson determined the shift was from high concentrations of ozone, and not due to exposure to light. Epson recommends Heavyweight Matte or Epson Photo paper for maximum lightfastness.
-if the "Photo Quality Glossy Paper" isn't new, it may be the paper's fault - it's been reformulated to deal with this problem. See http://www.luminous-landscape.com/1280.htm & http://www.wilhelm-research.com/
5) printer settings - make sure that when you print B&W prints, set the printer to "B&W". Colors can appear when "color" is selected, and the color printing mimics B&W
6) protective sprays - Luminos http://www.lumijet.com has a spray called "Imagesheild" to protect prints. $18/ can. Quoting their website: "LUMIJET IMAGESHIELD will become your favorite print protector. It*s a convenient ozone-friendly low-odor aerosol spray that significantly improves wet-fastness. In addition to protecting delicate inkjet images from moisture, it also shields images from fingerprints and harmful UV rays. This unique product will not only help extend useful print life, it is totally transparent; preserving the original print surface characteristics. From dead matte to glossy, or from canvas to silver foil, you*ll never know it*s there."
-Krylon clear acrylic spray Use many light coats. If you really wet the surface, the paper can become translucent.
-Johnson's Paste Wax. It works on some inks and papers, not all. It does offer some protection against UV. It does protect against moisture, finger prints, dust and scratches. It does change the finish. If the wax causes the ink to smear a fixative spray will usually stop that. Test it for several weeks before putting it on an expensive display.
Comments: Do not use the printer for archival printing. For long lasting prints use a silver halide digital printer (or dye sub) or traditional dark room processing.
Ink-jet prints have never been designed for long term preservation. the inks used in those printers are made of dyes, which are very sensitive to U.V electromagnetic waves present in fluorescent lamps and sun rays. Dyes are not chemically fixed to the fibers, so can cause the smudging problems. The use of a fixative to stabilize the colour is only a temporary and partial solution: the polymers spayed on the surface of the print might partly protect the dyes by filtering some U.V rays but doing so, will tend to yellow with time and give a yellowish cast to your images. Digitizing images is not a complete longterm solution either. For best preservation of images, save many copies on CDs, and check them often for deteroiration and update the formats as needed to be sure they are always accessible (tiff uncompressed files) and keep it in a cool place, with low humidity (30% RH), or rent a place in a specialised data keeping system firm.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
You are absolutely right, and most of the time we behave just as you describe us. Patient, etc. One purpose of my tirade was to point out that there are differences between today and yesterday that give ALL of us choices to make. I know a high school student who failed the year, because he wanted to punish his parents. When he told me what he had done, I responded that I knew a young man just like him when I was in high school. His solution was to get straight A's so he could become more quickly independent. Science is our own, very human, endeavor. Those of us who do science must constantly work to remove the temptations to embellish our results, to twist the statistics and to 'jump' to conclusions. But, and here's where I differ perhaps slightly. Sometimes, there is a good reason to yell! My wife hates my streak of sarcasm! But I was raised on the stuff. When I did something 'dumb,' everyone who loved me, joined in the derisive repair. I learned to think at my dinner table before I was 10. I did not like to be the but of everyone's attention. My Mom taught, then I taught. I only stopped, because no one [too few] wanted to learn any more [bad generalization]! Go to college today, and what do you find? Young people unprepared for the rigor. Young people unprepared for the intellectual dance - as we used to call it. Young people who don't know what they want to do and are unwilling to work to find out. "They only do what they are taught!" This is my mantra when I look for explanations in the mirror. My older son had to learn - for the test - to name NH3 as "ammonium", because his chemistry teacher would not believe the Handbook of Chemistry and Physics. My chemistry teacher in public high school took chemistry courses at Rutgers every third summer. He was the expert in chemistry. My physics teacher had a PhD in the subject. My biology teacher went on to be the chair of Physiology at a large Southern medical school. If I wanted to complain about a professor at Lehigh, the Dean would show me a list of nearby schools that had openings. The attitude was clearly communicated. The greatest lessons in life are those that teach one to manage the worst parts of it. If I failed a course, it was MY failure - and it was at least MY problem! I wanted to be what I am, once I figured everything out. The only advice I ever received, when I was having problems with learning, was, "Study harder." My Mom just kept on telling me to keep on trying as hard as I could. If I were young now, I'd be in group therapy and zapped on some pharmaceutical, if the system had its way. Given my experience, I hope it is not difficult to understand why I might not appear to understand the problems of others. BUT! If I see a young person in science who is poorly informed about how it is done, then I will act in that person's behalf even if I am thought, and reported, to be insensitive. When I was a graduate student, we were all ordered to write faculty/course evaluations. I did. I wrote analyses of each course, and every one of them was critically positive. Not one word of my analyses were printed. If I had demonized one of my professors, I would have been writing novels today. The people who ordered the writings were ex-faculty administrators who were pandering to the new consumer mentality in education. If I had gone to my undergraduate department chair to complain about a grade, he would have told me that I was wasting his time. Five years later, there were lines of boomers outside of department offices. The generation of manipulators had arrived! While they were standing in line, I was learning. While I was learning, they were just getting warmed up! I have had ideas snitched! Sometimes I even gave them away. On the other hand, I have also had the other side of the experience. For six months of learning followed by 30 minutes of consultation and work, I had the good fortune to become a molecular biologist. For that effort, the individual to whom I gave my best remembered my part - even after he had spent years doing the work - and included me in the author list for a gene(TERE1) that mapped to Chromosome 1 and appeared in print in a Nature publication. He did NOT have to do that. There was NO obligation nor agreement. I helped because he asked. People are basically good, especially scientists, and I always start with TRUST, and I always will. You are still absolutely correct.
Regards for YOUR sensitivity,
Fred Monson
} ---------- } From: Sousan Abolhassani } Sent: Thursday, June 28, 2001 8:07 AM } To: Paul Webster } Cc: MSA listserver submission } Subject: Re: long-term storage OR what has happened? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Paul, } } The point about this server is that it is used by } a very heterogeneous public from which a non-exclusive } list is the following: } } students, } professors, } scientists/ } researchers } technical staffs } suppliers } producers } specialists } non-specialists } etc. } etc. } etc. } } and all these from so many different backgrounds with so } many different problems and even from so many different countries... } That if we want to teach all of them to behave as we expect } before they post a query, the server could go to early retirement. } } I guess the philosophical attitude that one should have is to } really try and understand both sides of the discussion. } } 1- on one hand the trust that professor Monson is demanding } is not there because our users are a small sample of our } society. How many times have we heard that people have had } their ideas picked by others and their publications stolen } by others and .... in the very scientific community that most } of us hoped to be the purest part of the society. Still } the scientists are generally speaking very sincere people. } So if someone is not having trust to others, it's the } consequence of what they have observed in their environment. } } 2- I do not know if Rachel is a student (may be not), and if she } really was not trusting or just did not think to write the reason and } so on, so we do not discuss the case on a specific way. } But talking about going to the library, there are labs } where the students have really not access to the books and there } are places where superiors do not give the support the students } and other researchers' need, or simply there are not } enough books available. } } So there are always reasons for the way people behave } and the more our society goes towards a very materialistic and } "win whatever the situation" attitude, the more the arguments } of professor Manson will be applicable because the more people } will behave in the way he is not happy about, and I understand } and apreciate his point. } } All that apart, it is better to listen to people,as often things } that are not to be done are not published and are only "known". } Of course provided that answers are given generously, as arrogance } for giving is so destructive, even for donator. } I therefore agree with the fact that we should just be able to } ask the question and someone will react to give us an answer. } } I think that professor Monson has evoked a point that is of } great importance and that is the human aspect of all we do, as } "human" is what we are hoping to be at a first place. } That needs sensitivity... and continuous effort. } } Regards, } } Sousan } } } Paul Webster wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Colleagues, } } } } The messages from Professor Monson in reply to a question about } long-term } } tissue storage makes me wonder what has happened to this listserver? } } } } His message was highly entertaining and even if the poster of the } original } } message may have felt "picked on", there was no reason to expect him to } post } } an apology, no matter how informative it was. } } } } Far too many posters use this listserver as a way of finding a quick fix } to } } simple solutions to technical problems that can be easily researched in } text } } books. } } } } Often these messages contain little background information on the } technical } } problem under investigation. Even so, many of these questions get full } } attention and detailed replies, even if the replies are not posted on } the } } main list for public view. } } } } What has happened to the free and open discussion of issues and } protocols } } that we used to have? Are people so afraid of being accused of being } } incorrect that they do not post their replies openly? Or maybe people } are } } afraid of being accused of being impolite. } } } } I look forward to controversy and hope that my comments are read } carefully } } and replied to with vigor. Only in this way will I be able to learn new } } thing, and this is the main reason I still subscribe to this listserver. } } } } If I am factually wrong about something, tell me. It is better to find } this } } out on the listserver than in a more professional (or cruel) } environment. } } } } As for discussion subjects: why was there no comment about the } unadvisable } } suggestion made about storing samples in a frozen state? Would anyone } } working with EM really choose to store their samples at -20 degrees } before } } processing for TEM? } } } } Where is all the discussion about extraction that can occur when storing } } samples in alcohols or buffers, or on the contamination that I know } grows in } } everyones old cacodylate buffers? } } } } Do we not comment because we know it has all been published in } textbooks? } } } } Dear Profesor Monson, do not feel bad about improving our general } knowledge } } and our professional etiquette. I for one support your views and find } your } } comments useful and entertaining. Please keep up your submissions. } } } } Best regards, } } } } Paul Webster } } } } Paul Webster, Ph.D. } } Scientist II and Director } } Ahmanson Center for Advanced EM & Imaging } } House Ear Institute } } 2100 West 3rd Street } } Los Angeles } } CA 90057 } } } } pwebster-at-hei.org } } p: 213 273 8026 } } f: 213 413 6739 } } http://www.hei.org/research/depts/aemi/aemi.htm } }
} ---------- } From: Gareth Morgan } Sent: Thursday, June 28, 2001 8:18 AM } To: Paul Webster; MSA listserver submission } Subject: Re: long-term storage OR what has happened? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Colleagues } } I agree wholeheartedly with Paul in his comments regarding the mails from } Frederick Monson - no need for apology, interesting and informative } replies. } } Higher education seems to be locked in an apparently paradoxical situation } with students being expected to 'take responsibility for their own } learning', the use of Student Centred & Problem Based Learning etc. The } paradoxical part is that they do not have the appropriate intellectual } tools to be able to carry out such assignments. Frustration with their lot } then spills out over tutors who are expected to take responsibility when } things go wrong (but mostly no feedback when things work). } } Ask me a question and I will give you the most appropriate answer I can. } The answer may be wrong of course, possibly because I have an incorrect } understanding of the concepts involved, possibly because I was asked the } wrong question. If I respond to a question with a question of my own it is } because I am trying to seek clarification of what EXACTLY is being asked. } That, as I see it, is how we work in science. } } Paul says - "If I am factually wrong about something, tell me. It is } better } to find this out on the listserver than in a more professional (or cruel) } environment." } } I agree absolutely here too - it is easier to fight tooth and claw here } and } come to some agreement or compromise than if we are addressing 500 people } at a conference and one of them asks a simple question to which we have no } answer. } } My appeal to those posing questions is to give as much background } information as possible and to respond clearly to questions on } clarification. } } My appeal to those answering is to identify clearly what is hearsay "in my } experience" "I got that straight from a guy in the pub last night". } Identify also the context of the answer/solution to the question/problem } and refer to published data whenever possible. } } I look forward to a lively, ongoing debate (even though I am going for my } summer break tomorrow!). } } } } } Med v?nliga h?lsningar/With best wishes } } Gareth } } "Close your eyes and look. What you saw at first is there no more; and } what } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } http://www.ki.se/biomedlab e-mail Gareth.Morgan-at-impi.ki.se } } Tel +46 8 728 3734 } Fax +46 8 728 3688 } } Gareth Morgan MPhil MSc FIBMS, } Institutionen f^r Mikrobiologi, Patologi och Immunologi(IMPI), H5, } Karolinska Institutet, } Avdelningar f^r biomedicinsk } laboratorievetenskap och } biomedicinska ?mnen, } Lindhagensgatan 92, Box 12773, } S 112 96, Stockholm } Sverige } } OBS! Bes^ksadress: Lindhagensgatan 92 } NB! Visiting address:Lindhagensgatan 92 } }
} ---------- } From: Gareth Morgan } Sent: Thursday, June 28, 2001 7:45 AM } To: Christine Edly; Microscopy-at-sparc5.microscopy.com } Subject: Re: immersion oil remover? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi } } I have used the 'orangey' smelling solvent under the brandname of } Histoclear. It is supposedly extracted large scale from citrus peel and is } the composed of the 'zest' that comes out as a spray when we peel oranges. } } I couldn't comment on its use for removing immersion oil as you describe. } I } have only used it as a xylene substitute for tissue processing tissues and } also for clearing stained sections prior to mounting. } } It is said to be safer than xylene but some of the people I worked with at } the time complained of headaches so we stopped using it despite never } proving that it was the cause of the headaches. } } Hope this helps! } } } } At 17:42 2001-06-27 -0500, Christine Edly wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Greetings! } } } } I'm in search of something that will remove immersion oil from slides of } } } } blood smears (no coverslips) without damaging the specimens. A friend } } told that such a chemical exists, but could not remember the name. It } } supposedly smells like oranges, and xylene was once used for the same } } purpose. If you have any ideas as to the name of this mystery chemical, } } } } I'd sure appreciate it. } } } } Regards, } } Christine Edly } } University of Oklahoma } } } } } Med v?nliga h?lsningar/With best wishes } } Gareth } } "Close your eyes and look. What you saw at first is there no more; and } what } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } http://www.ki.se/biomedlab e-mail Gareth.Morgan-at-impi.ki.se } } Tel +46 8 728 3734 } Fax +46 8 728 3688 } } Gareth Morgan MPhil MSc FIBMS, } Institutionen f^r Mikrobiologi, Patologi och Immunologi(IMPI), H5, } Karolinska Institutet, } Avdelningar f^r biomedicinsk } laboratorievetenskap och } biomedicinska ?mnen, } Lindhagensgatan 92, Box 12773, } S 112 96, Stockholm } Sverige } } OBS! Bes^ksadress: Lindhagensgatan 92 } NB! Visiting address:Lindhagensgatan 92 } }
I bet we would all like to see some results when you get them.
Regards,
Fred Monson
} ---------- } From: Shea Miller } Sent: Thursday, June 28, 2001 4:58 PM } To: microscopy-at-sparc5.microscopy.com } Subject: plant immunofluorescence-thanks } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks so much for all the info on protocols for plant } immunofluorescence... we tried a trial run today on some immature soybeans } embedded in paraffin, and got some positive results following the general } methods of Tobias Baskin. You have all given great advice, and we have } lots to work with to refine our immunoassay. } } thanks so much } shea } } } } Dr. S. Shea Miller } Agriculture & Agri-food Canada } Eastern Cereal and Oilseed Research Centre } Central Experimental Farm } Ottawa, Ontario } Canada K1A 0C6 } Phone: (613) 759-1760 } Fax: (613) 759-1701 } Email: millers-at-em.agr.ca } } }
The classic blood stains are the dried salts of cationic(methylene blue) and anionic(eosin) dyes dissolved in methanol. So, acid will remove one and base will remove the other, and the aromatic organic is what we no longer want to use. How about a non-ionic detergent? Is that what's in the "Orange" mystery?
Well here's my offering for something 'orangy'. I acquired it a Home Depot. "Commercial ZEP Citrus All Purpose Cleaner". The Directions say that you can use it full strength or diluted. Probably doesn't have any of THOSE aromatics. Nothing on the label. Looks like a typical proprietary scientific mixture. Just like "Histoclear" and "Paraplast." [Why does that period look so out of place?] "Removes grease, oil and soils [Sic!] from concrete, metal and other hard surfaces."
Glass is hard unless one has a very long perspective. I wonder. If I had some old slides, I could try it. Full strength or a dilution series? Ten-fold or two-fold? Water or alcohol as a vehicle? Wonder whether "triphosphate" would work. Did a fine job on the driveway. Time for lunch! I'll stop by Home Depot on the way back.
Respectfully submitted in partial fulfillment of requirements for.......,
Fred Monson [Is he this way all the time? Only when awake.] } ---------- } From: Christine Edly } Sent: Wednesday, June 27, 2001 6:42 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: immersion oil remover? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings! } } I'm in search of something that will remove immersion oil from slides of } } blood smears (no coverslips) without damaging the specimens. A friend } told that such a chemical exists, but could not remember the name. It } supposedly smells like oranges, and xylene was once used for the same } purpose. If you have any ideas as to the name of this mystery chemical, } } I'd sure appreciate it. } } Regards, } Christine Edly } University of Oklahoma } }
I am very grateful to everybody for your help with regards to finding suitable TEM grid film for small particles. The number of responses were really overwhelming. Special thanks to those who were willing to send me grids to try out. Thank you all once again.
Hi, everyone. I am posting this for someone else. However replies can be either posted on the server or sent to my email address directly. I will forward all to her. Thank you in advance.
Hong
Hi Hong,
Okay... here is a paragraph or so about what we are trying to see that you can paste onto the list serv.
We are attempting to visualize the polysaccharide capsule of Neisseria meningitidis serogroup B by electron microscopy. The capsule is (alpha2-8)-linked polysialic acid with about 200-300 residues per polymer. We have tried staining the negatively charged capsule with Ruthenium red, followed by embedding, sectioning, and counterstaining. However, it appears that only the inner and outer membranes were stained and not the capsule (we did capsule + and capsule - controls). In a previous paper that effectively stained capsule of E. coli K5, the cells were washed with capsule-specific antibody before fixation to stabilize the capsule. We have not attempted the procedure with the antibody stabilization yet. Please let me know if this would be recommended or if there are any other suggestions. Thank you.
Hi again; we are having a wee bit of trouble distinguishing our signal from the autofluorescence in our immunoassay on immature soybean seeds/seed coats. So far, we have used a light stain of toluidine blue, but I'm not completely happy, and am considering Evans Blue instead. What do others use to quench the yellow autofluorescence (our Ab is FITC labelled) in plant tissues, and when do you actually apply the counterstain? Before the Ab incubations, or after??
thanks so much again in advance shea
Dr. S. Shea Miller Agriculture & Agri-food Canada Eastern Cereal and Oilseed Research Centre Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 Email: millers-at-em.agr.ca
I never heard that before. After 36 years of marriage, and I find this out NOW?
Fred Monson } ---------- } From: Nicol Aitken } Sent: Friday, June 29, 2001 10:43 AM } To: 'Monson, Frederick C.' } Subject: RE: Today's Real Humor } } I'll answer e, even though strictly speaking osculation need not occur! } Nick } } -----Original Message----- } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] } Sent: Thursday, June 28, 2001 1:30 PM } To: 'Microscopy Listserver' } Subject: Today's Real Humor } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Question from an old exam given to students who demanded MORE MULTIPLE } CHOICE! } } Which of the following is NOT involved with human reproduction? } } a. transcription } b. replication } c. translation } d. osculation } e. none of the above } } Answer tomorrow. } } } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI) } Schmucker II Science Center (Room: SS024(Basement)) } South Church Street } West Chester, PA, 19383 } MailDrop: Department of Geology/Astronomy } Phone: 610-738-0437 } Fax: 610-436-3036 } email: fmonson-at-wcupa.edu } Please call before visiting. } }
Perhaps I'm missing something from the biological side with which I am not familiar. However, for my purposes, it has always been very easy to remove immersion oil using a non-polar organic such as one of the following: hexane, iso-octane, VM&P Naphtha (Varnish Maker's and Printer's Naphtha from the hardware store), "petroleum ether" (not really a C-O-C ether but a mixed alkane distillate), ligroine, etc.
These have the following attributes - they are extremely low in viscosity so they penetrate any porosity quickly and dissolve the oil; they evaporate quickly so that any residual oil quickly becomes apparent, they do not swell or dissolve most mounting materials (limonene - the environmentally friendly base of many "green" solvents, is a potent stripper of polymers); and their high vapor pressure makes them easy to eliminate prior to SEM examination. I have routinely used immersion oil to temporarily coverslip geological thin sections for optical microscopy prior to microprobe work and then removed the oil with these solvents. Several drops in succession, less than a milliliter total, is sufficient to clean one slide.
John Twilley
Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Please post informational responses to the server! I've been curious about } this, too but too lazy to look into it - the dishwashing cleansers sold in } "granola" stores (organic, non-toxic to the environment, etc.) are also } citrus-oil based (and at least one has almond oil; not sure if that is } just for the odor). } } Tamara
As we have been instructed for the past twenty years, nothing is NOT involved with that stuff.
So, 'e' is the correct answer. My students learned to hate multiple choice as much as I did.
Regards to all, with advice to my son. Always brush your teeth first, so the kisses are sweet. And he asked, "Mint or wintergreen?"
Have a nice weekend and remember, we can all get away with this because someone once worked on the 4th of July!
Fred Monson
} ---------- } From: Monson, Frederick C. } Sent: Thursday, June 28, 2001 1:30 PM } To: 'Microscopy Listserver' } Subject: Today's Real Humor } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Question from an old exam given to students who demanded MORE MULTIPLE } CHOICE! } } Which of the following is NOT involved with human reproduction? } } a. transcription } b. replication } c. translation } d. osculation } e. none of the above } } Answer tomorrow. } } } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI) } Schmucker II Science Center (Room: SS024(Basement)) } South Church Street } West Chester, PA, 19383 } MailDrop: Department of Geology/Astronomy } Phone: 610-738-0437 } Fax: 610-436-3036 } email: fmonson-at-wcupa.edu } Please call before visiting. } }
Here's an old list, checked and amplified, that came to me from "Microscopy Today" I have checked all of these URL's
www.home.earthlink.net/~veilcs/ (EM+ special in Philips) (west Coast - USA) www.southernmicro.com (Georgia) (Research LM) www.dscoptical.com/services.htm (Boston?) (LM) www.caleywhitmore.com (Boston?) (LM) www.opticalexpertise.com (NY) (connection failure?) www.mwrn.com/product/microscope/repair.htm (LM and EM) www.sciscope.com/field-service.htm (30+ states) (up to Advanced LM) www.svms.com/services/index.html (CA) (LM) www.meyerinst.com (TX) (up to advanced LM) www.microresource.com/services2.htm (IL) (LM) www.allometrics.com/microscope_serv.htm (LA)(LM) www.labworksinc.com (San Francisco) (Limited to specialized LM scope accessories) www.mikronet.com (San Diego) (general LM services) www.uams.edu/oas/OES/oesilab.htm (AR) (University of Arkansaw LM support facility) www.uni.edu/pur/mms/equipment.html(U. N. Iowa) (Wide range of equipment including EM's) www.dominionmicroscope.com/services.htm (Richmond, VA) (LM and associated equipment) www.eosltd.co.uk/ (UK.COM) www.emsys.co.uk/ (UK.COM) www.hii.hitachi.com/svcem.htm (Hitachi) www.leo-usa.com/ (LEO) www.feic.com/products/index.htm (FEI - Philips) www.jeol.com/locations/service/service.html (JEOL Service, USA)
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
Some faculty here are entertaining the idea of developing a confocal core facility. There is currently a single confocal microscope on campus in an individual PI's lab and it is not as available to others as they might like due to that lab's workload, location, etc.
I have been invited to join the group investigating the possibility of a second, more general access microscope, probably supported by several departments. I am currently the person responsible for the EM lab on campus, we have a TEM and an SEM, plus most of the standard prep. equipment. We also do some digital imaging. The EM business is not booming, save for a few hardcore users.
Since I am one of the 'Mr. Microscopes' here, I would like to join this group with a running start. I ask for your sage advice regarding the potential good and bad of this proposal and any comments you might offer on the state of confocal and such a lab.
I know a lot about microscopes, less specifically about confocals, but I could learn. If the microscope was put into the existing EM lab, I have a separate room, bench space, hoods etc. Just about anything needed to do microscopy. The only drawback would be that we are in a different building than most of the users. Its not a bad walk, but how big a deal is it to prepare specimens away from one's own lab? It would be pretty fair for the multiple users, since none of them are all in the same building either, they would all have to do some traveling to get here.
I am pretty sure there will be the usual squabbling that goes along with central facilities, like who gets to use it if there is competition, who will pay for it, etc. Is that likely to be any worse then with any shared equipment? Am I asking for trouble to have it added to the EM Lab?
Do you have an experience with the kind of day to day issues that a lab like this might generate. Are there lots of supplies that need to be ordered and kept on hand? Is the equipment reliable and are there any special features of one particular type of instrument that would make it more appropriate for such a general use facility vs. something used by an individual lab?
Your comments will be carefully considered and as always, greatly appreciated.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
The department I worked for took a different approach to the students not "ready for the dance". Every Thursday night the lab was open a faculty member from assistant professor to department head was there from 7:00 pm until at least 11:00 pm to help them with what ever problems they had not just Ag Engineering courses. I worked a research computer programmer and we didn't teach any programming courses and I had not teaching duties at all but any student could come to me any time with a computer problem and the same was true for any member of the staff or faculty for any problem the students had.
No grad students taught courses or labs.
The policy paid off real well. I don't remember when one of our students didn't get a job after graduation and many have gone on to very good things. This years senior design class is applying for patent on part of their class project.
I realize that farm kids are a good deal better prepared for life than most students but their education is usually a good deal poorer in science and math than the kids from the city and they have a good deal of catching up to do.
You are short changing your students if you just present the knowledge and walk away.
I realize that the approach we used is difficult to use in departments with large number of students. Turning education into a mass market commodity is IMO much of what is wrong with it.
Gordon
Gordon Couger
Stillwater, OK
----- Original Message ----- } From: "Monson, Frederick C." {fmonson-at-wcupa.edu} } } } You are absolutely right, and most of the time we behave just as you } describe us. Patient, etc. One purpose of my tirade was to point out that } there are differences between today and yesterday that give ALL of us } choices to make. I know a high school student who failed the year, because } he wanted to punish his parents. When he told me what he had done, I } responded that I knew a young man just like him when I was in high school. } His solution was to get straight A's so he could become more quickly } independent. } Science is our own, very human, endeavor. Those of us who do } science must constantly work to remove the temptations to embellish our } results, to twist the statistics and to 'jump' to conclusions. But, and } here's where I differ perhaps slightly. Sometimes, there is a good reason } to yell! My wife hates my streak of sarcasm! But I was raised on the } stuff. When I did something 'dumb,' everyone who loved me, joined in the } derisive repair. I learned to think at my dinner table before I was 10. I } did not like to be the but of everyone's attention. My Mom taught, then I } taught. I only stopped, because no one [too few] wanted to learn any more } [bad generalization]! Go to college today, and what do you find? Young } people unprepared for the rigor. Young people unprepared for the } intellectual dance - as we used to call it. Young people who don't know } what they want to do and are unwilling to work to find out. "They only do } what they are taught!" This is my mantra when I look for explanations in } the mirror. My older son had to learn - for the test - to name NH3 as } "ammonium", because his chemistry teacher would not believe the Handbook of } Chemistry and Physics. My chemistry teacher in public high school took } chemistry courses at Rutgers every third summer. He was the expert in } chemistry. My physics teacher had a PhD in the subject. My biology teacher } went on to be the chair of Physiology at a large Southern medical school. } If I wanted to complain about a professor at Lehigh, the Dean would show me } a list of nearby schools that had openings. The attitude was clearly } communicated. The greatest lessons in life are those that teach one to } manage the worst parts of it. If I failed a course, it was MY failure - and } it was at least MY problem! I wanted to be what I am, once I figured } everything out. The only advice I ever received, when I was having problems } with learning, was, "Study harder." My Mom just kept on telling me to keep } on trying as hard as I could. If I were young now, I'd be in group therapy } and zapped on some pharmaceutical, if the system had its way. } Given my experience, I hope it is not difficult to understand why I } might not appear to understand the problems of others. BUT! If I see a } young person in science who is poorly informed about how it is done, then I } will act in that person's behalf even if I am thought, and reported, to be } insensitive. } When I was a graduate student, we were all ordered to write } faculty/course evaluations. I did. I wrote analyses of each course, and } every one of them was critically positive. Not one word of my analyses were } printed. If I had demonized one of my professors, I would have been writing } novels today. The people who ordered the writings were ex-faculty } administrators who were pandering to the new consumer mentality in } education. If I had gone to my undergraduate department chair to complain } about a grade, he would have told me that I was wasting his time. Five } years later, there were lines of boomers outside of department offices. The } generation of manipulators had arrived! While they were standing in line, I } was learning. While I was learning, they were just getting warmed up! } I have had ideas snitched! Sometimes I even gave them away. On the } other hand, I have also had the other side of the experience. For six } months of learning followed by 30 minutes of consultation and work, I had } the good fortune to become a molecular biologist. For that effort, the } individual to whom I gave my best remembered my part - even after he had } spent years doing the work - and included me in the author list for a } gene(TERE1) that mapped to Chromosome 1 and appeared in print in a Nature } publication. He did NOT have to do that. There was NO obligation nor } agreement. I helped because he asked. People are basically good, } especially scientists, and I always start with TRUST, and I always will. } You are still absolutely correct. } } Regards for YOUR sensitivity, } } Fred Monson
with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder with a 4X marked on it.
Focus for images was done through the monocular eyepiece on the camera assembly.
I assume that my magnification though the microscope eyepiece would be 30X I assume that the magnification to the camera back would be 24X
Sorry if this seems like a silly question but I do not do much optical microscopy and got to thinking that it may be more than just the sum of the optics magnification.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
The House Ear Institute, a Los Angeles-based, non-profit research and education center dedicated to the hearing-impaired, has experienced tremendous growth over the past 50 years. We currently have an exceptional opportunity for a motivated and innovative self-starter.
You will support research projects in the cell and molecular biology of hearing. You will also perform advanced research procedures, such as immunoelectron microscopy and microwave processing, and maintain records. Requires a detail-oriented, flexible and enthusiastic team player with a BS in the biomedical field, as well as knowledge of TEM and SEM, specimen preparation for TEM and SEM, darkroom developing/printing, and digital image capture/manipulation. Must have efficient communication skills. Familiarity with electron microscopes, mechanical/electronic equipment, vacuum systems, and Unix, PC or Mac is ideal. Experience in cell culture, microbiology or protein purification is useful but not essential.
The Ahmanson Advanced Electron Microscopy and Imaging Center was recently established to perform independent research and provide support to a cell and molecular biology program in hearing research. The Center has a CM 120 BioTwin TEM and XL30 SFEG SEM. Specimen preparation equipment include ultramicrotomes for resin and cryosectioning, sputter coaters, vacuum evaporator, freeze-substitution, freeze-fracture and rapid freezing devices, a microwave processor, digital and stereologucal imaging capabilities and an SGI octane with 3-D reconstruction software.
The House Ear Institute, located in the downtown Los Angeles area, is a non-profit research and education organization dedicated to improving the lives of people who have hearing and hearing related disorders. The House Ear Institute is an Equal Opportunity Employer.
For consideration, please submit resume and cover letter to: House Ear Institute, Human Resources, 2100 W. 3rd St., Los Angeles, CA 90057-1922. FAX: (213) 483-8789. E-mail: jobs-at-hei.org.
When applying, please state that you saw this position advertised on the MSA listserver.
Regards,
Paul Webster
Paul Webster, Ph.D. Scientist II and Director Ahmanson Center for Advanced EM & Imaging House Ear Institute 2100 West 3rd Street Los Angeles CA 90057
this is some kind of follow-up question to Thearith's grid problem. We are looking at about 2 nm features in a polymer matrix. The features have very low contrast with respect to the all-carbon matrix due to the fact that they mostly contain O and a little Zn. For sample prep reasons we need to support the samples. Now: Are SiO and Si nitride coatings thin enough to still see such things ? The available thicknesses of these films seem very large compared to conventional C films.
TIA,
Andreas
************************************************* Dr. Andreas Taubert Dept. of Materials Science and Engineering 3231 Walnut Street University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
Not thinking today, in my previous message I made assumptions about total magnification being the sum of the objectives but should have been the product.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Yes, Fred, we published this list in Microscopy Today. We got it from a lady from Australia, whose name currently escapes me. Due to the interest in this topic, allow me to request that all readers contact companies that they know that does microscopy (all techniques)independent service. Ask them to contact me by email with the following: 1) Name of organization. 2) Contact info (tel. number and web site) 3) Products serviced 4) Area served (states, region, whatever) I will organize and publish the full listing, with Nestor's blessing, on this server. Best to all, Don Grimes, Microscopy Today
-----Original Message----- } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] Sent: Friday, June 29, 2001 12:23 PM To: 'service-at-jcnabity.com' Cc: 'Microscopy Listserver'
Here's an old list, checked and amplified, that came to me from "Microscopy Today" I have checked all of these URL's
www.home.earthlink.net/~veilcs/ (EM+ special in Philips) (west Coast - USA) www.southernmicro.com (Georgia) (Research LM) www.dscoptical.com/services.htm (Boston?) (LM) www.caleywhitmore.com (Boston?) (LM) www.opticalexpertise.com (NY) (connection failure?) www.mwrn.com/product/microscope/repair.htm (LM and EM) www.sciscope.com/field-service.htm (30+ states) (up to Advanced LM) www.svms.com/services/index.html (CA) (LM) www.meyerinst.com (TX) (up to advanced LM) www.microresource.com/services2.htm (IL) (LM) www.allometrics.com/microscope_serv.htm (LA)(LM) www.labworksinc.com (San Francisco) (Limited to specialized LM scope accessories) www.mikronet.com (San Diego) (general LM services) www.uams.edu/oas/OES/oesilab.htm (AR) (University of Arkansaw LM support facility) www.uni.edu/pur/mms/equipment.html(U. N. Iowa) (Wide range of equipment including EM's) www.dominionmicroscope.com/services.htm (Richmond, VA) (LM and associated equipment) www.eosltd.co.uk/ (UK.COM) www.emsys.co.uk/ (UK.COM) www.hii.hitachi.com/svcem.htm (Hitachi) www.leo-usa.com/ (LEO) www.feic.com/products/index.htm (FEI - Philips) www.jeol.com/locations/service/service.html (JEOL Service, USA)
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
Ok, I can't resist... I haven't seen anyone mention the way I learned to do it in 1962...
Cut lens paper into strips a bit wider and longer than the slide. Place an end of the paper over the end of the slide and add a drop or so of your favorite liquid to the paper on the slide...just enough to spread over the width of the slide. Grasp the edge of the paper and pull slowly down the length of the slide so that the excess liquid is wicked into the remaining paper as it is pulled across the slide. Try to run out of paper as you reach the other end of the slide.
As Ever, Orin Keplinger ----- Original Message ----- } From: "Tobias Baskin" {BaskinT-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, June 28, 2001 4:28 PM
Hi there fellow microscopists,
Here in the Pathology EM lab we do not do rocket science, but standard thin-section TEM of renal biopsies.
In the last two weeks though I have been getting some ka ka (technical term) on my grids from my UA stain. I have already made up two new batches of UA stain and use .45 micron filters just before use. Question is that all of the sudden I am getting UA stain artifact on my grids. I have not have any trouble since I have been here. I have changed everything, new staining plate, cleaned and scrubbed everything that I use for staining. I still get some UA artifact. Is this normal and should I just forget about it or is there something else wrong I am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I stain for usually 10-12 minutes. the grids before staining or clean, but after UA stain they are a bit dirty...
Any suggestions?
Thanks,
Eric A. Rosen UCLA Medical Center Dept. Pathology EM Lab
Fellow colleagues,I am in a dilema. I just found out that RMC(whom our lab had Maintenance Agreement on two MT2 and one MT2B) has been taken over by first Ventana and at a later date Biobeck? Later company is not offereing maintenance service contracts for "old" MT2 or MT2B. Is there any one out there that could refer me to someone who works on these great workhorses in EM? Thanking you in advance, Teresa
Listers: I've been following the discussion of lens cleaing and would like insert my two cents worth. I'm from far outside of generally biological tenor of this group; an industrial metallographer ( microscopy of metal and alloys). After using an oil immersion lens, I clean it with Kodak lens cleaner fluid after blotting up the excess oil with a lens tissue, also Kodak. After 30 years of use, my lenses are clear and clean with no apparent deterioration. I did use xylene at first but was dissuaded by knowledgeable microscope salesman.
Sam Purdy Technical Center National Steel Corp. Trenton MI
Hello, Many thanks to everyone for their helpful comments on the Electrscan E3 SEM. We are still having vacuum problems in wet mode only. It works fine in High vacuum mode though. We checked the diffusion pumps, tried moving a rotary pump closer to the column, but no improvement. We are slowly changing o-rings to see if that helps.
Does anyone have any detailed repair manuals or diagrams with o-rings for this instrument? We have the customer maintenance manual, but it is rather limited. It mainly covers how to change apertures and doesn't go too deep into troubleshooting the vacuum system.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I don't use this scope but I could venture the following guestimates.
If the photo system and light path does not include a separate photo objective (photo eyepiece) then the film plane magnification is the objective x 4. This would then be 20 x 4 = 80X.
Usually, with a 35mm camera body, the system is set up to be 1:1 with the oculars. This would mean 20 x 10 = 200X. If this field of view is different with the 4x5 cone, then there is some intermediary lens element. Usually, it is a 2.5X objective. For photo work, the oculars won't make any difference.
Perhaps some Optiphot experts can help you better.
gary g.
At 02:16 PM 6/29/2001, you wrote:
} Need some help trying to figure the magnification of some photos taken with } the following: } } Nikon Optiphot-Pol Microscope } CF eyepiece 10X } CF objective 20X } } with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder } with a 4X marked on it. } } Focus for images was done through the monocular eyepiece on the camera } assembly. } } I assume that my magnification though the microscope eyepiece would be 30X } I assume that the magnification to the camera back would be 24X } } Sorry if this seems like a silly question but I do not do much optical } microscopy and got to thinking that it may be more than just the sum of the } optics magnification. } } Thanks } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu
We have found staining artifacts to be one of the most frustrating, intermittent mysteries of all. A while back we began having trouble with UA and Pb contamination where no trouble had occured before---same stains, same processing, same everything, but suddenly there was goop everywhere. We mixed new stains, changed all of our 0.2 um filters, spun everything down, and filtered again, used NaOH religiously, held our breath while staining, etc., etc., etc. Finally, the only thing we could think of was water, so we called in our water system maintainer (let's just call them "Hey Mystery Man!!!!) who had been servicing our reverse osmosis, deionization system forever.
Well, they said, your system's shot. The diaphragm's bad, the cartridges are way expired, and the filter is one big bucket of rust. You need to maintain this better. We informed them that we paid them a princely sum to maintain it for us and what the h... were they doing every month when they came in to fuss with it?
Turns out that they thought we owned the system and had only been swapping out some tank or other, and they assumed we were doing the rest. When we produced documentation of our service contract on their leased system that we had been paying on since about 1735 A.D., there was an awkward pause. Then we fired them.
We purchased a Millipore tabletop water purification system that solved our problems overnight. It's a small, low-volume gizmo that produces a few litres of ultra-high purity water a day. Costs about $1200, as I recall, and requires annual filter changes to the tune of $200-300.
Moral of the story: Check the water when all else fails. Worked for us, anyway. Couldn't hurt to check.
We have no stake in Millipore whatsoever, except as relieved users of one of their products.
Randy Tindall Electron Microscopy Core University of Missouri
-----Original Message----- } From: Eric To: 'Microscopy Listserver' Sent: 6/29/01 5:24 PM
Hi there fellow microscopists,
Here in the Pathology EM lab we do not do rocket science, but standard thin-section TEM of renal biopsies.
In the last two weeks though I have been getting some ka ka (technical term) on my grids from my UA stain. I have already made up two new batches of UA stain and use .45 micron filters just before use. Question is that all of the sudden I am getting UA stain artifact on my grids. I have not have any trouble since I have been here. I have changed everything, new staining plate, cleaned and scrubbed everything that I use for staining. I still get some UA artifact. Is this normal and should I just forget about it or is there something else wrong I am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I stain for usually 10-12 minutes. the grids before staining or clean, but after UA stain they are a bit dirty...
Any suggestions?
Thanks,
Eric A. Rosen UCLA Medical Center Dept. Pathology EM Lab
I met some problems during I prepared the samples for GaN/sapphire plan-view using tripod wedge technique. After polishing, the sample was thinned by pips in one side. However, The other side usually was covered by copper, as a result the sample became dark. It seems the unthinned side should be covered something to prevent sputtered materials from growing on it. Do you have some knowledge on it?
Thanks in advance.
Best regards.
Feng ********************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
Mica! 1stly core drill a 3mm disc. Then use sharp tweezers to peel a thin piece from dozens or even hundreds of layers in the mica disc depending how good you are to separate them. Anyway you just need one thin piece to cover the other side, and you may be able to reuse that thin piece.
********************************* Chaoying Ni, PhD Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
On Sat, 30 Jun 2001, Feng Wu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } } }
Dr. Feng Wu wrote: =============================================== I met some problems during I prepared the samples for GaN/sapphire plan-view using tripod wedge technique. After polishing, the sample was thinned by pips in one side. However, The other side usually was covered by copper, as a result the sample became dark. It seems the unthinned side should be covered something to prevent sputtered materials from growing on it. Do you have some knowledge on it? ================================================= There are two materials that to the degree we have been able to determine, are essentially equivalent for this purpose: One is called Microshield™ and the other Lacomit™.
SPI Supplies has offered Microshield for this purpose and more information about its use and application can be found on the SPI Supplies website at URL http://www.2spi.com/catalog/chem/microshield.html
Disclaimer: SPI Supplies has not done any careful analysis to prove that Microshield and Lacomit are identical, only that for this particular TEM application, they do seem to be equivalent. If there is information to the contrary, we would appreciate hearing about it. Suggestion: Use in conjunction with the Microshield remover.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ===============================================
} Need some help trying to figure the magnification of some photos taken with } the following: } } Nikon Optiphot-Pol Microscope } CF eyepiece 10X } CF objective 20X } Hi Roy,
If you still have access to the microscope, put a specimen on the stage which has a large feature, one you can accurately measure. Use the lowest power lens available. Remove the film cassette from the camera and stretch a piece of Scotch Magic tape across the film plane. Focus on the specimen, lower the room lights and measure the size of the image on the scotch tape, which will now be clearly visible. You could also measure some feature on the original slide and compare it to the photograph.
I would guess the mag factor is 80x, but you can confirm that using this process.
Dave Burton Optical Specialist University of Washington
Dr. Andreas Taubert asked: ============================================= this is some kind of follow-up question to Thearith's grid problem. We are looking at about 2 nm features in a polymer matrix. The features have very low contrast with respect to the all-carbon matrix due to the fact that they mostly contain O and a little Zn. For sample prep reasons we need to support the samples. Now: Are SiO and Si nitride coatings thin enough to still see such things ? The available thicknesses of these films seem very large compared to conventional C films. ============================================= If the thinnest thickness of the nitride should still be too much, which is presently 30 nm, but soon to be reduced to the 10-20 nm range, the present silicon nitride membrane grids can be furnished "holey", that is, with a matrix of holes that are about as small as you want them, in just about any arrangement that you might want them. In that kind of situation, there is no nitride underneath the sample being analyzed, so long as you are looking "through" the holes. The only carbon (or oxygen) present is that which is coming from the sample. The typical compliment of holes approximately doubles the cost of the grid, because it is a separate operation done by micromachining techniques. This is explained in greater detail at URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
Disclaimer: SPI Supplies offers silicon nitride membrane window grids, both with and without the holes.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==============================================
The ion beam forms a cloud of debris, which settles on the reverse side of the specimen and this needs to be covered. A double beam ion gun system avoid that problem because the specimen is thinned on both sides simultaneously. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, June 30, 2001 5:06 PM, Feng Wu [SMTP:fwu-at-bgumail.bgu.ac.il] wrote: } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } } in one side. However, The other side usually was covered by copper, as a } result } the sample became dark. It seems the unthinned side should be covered } something } to prevent sputtered materials from growing on it. Do you have some knowledge } } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } **********************************************
i am trying to find a way into the literature on experimental techniques appropriate to study 511 photon detection and capture vs properties of scintillating materials. Is there a standard way to do this?
Can someone point me at a good place to begin a literature survey?' sorry, my question is not better phrased, but I am just beginning to look at this?
sterling
Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
Feng, the trick is not to have a hole in your specimen where the ions can go through and back-sputter the Cu on to your sample. So make your specimen big enough to cover up the hole in the support grid and stop ion milling immediately when a hole appears. The problem will go away to some extent if you use the two-sided specimen holder. Alternatively, use an inert grid and dissolve the Cu with an acid after specimen prep. (Most acids are unlikely to affect GaN or sapphire.)
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } }
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone." Marconi Optical Components Limited Registered in England No. 4113798 Registered Office: One Bruton Street London W1J 6AQ
} From reading posts on this listserver I gather the problem is usually related to a failure of the water purification system. If you use purified water why not make some UA up in double distilled water as a check.
Dave
On Fri, 29 Jun 2001 15:24:31 -0700 Eric {biology-at-ucla.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there fellow microscopists, } } Here in the Pathology EM lab we do not do rocket science, but standard } thin-section TEM of renal biopsies. } } In the last two weeks though I have been getting some ka ka (technical } term) on my grids from my UA stain. I have already made up two new batches } of UA stain and use .45 micron filters just before use. Question is that } all of the sudden I am getting UA stain artifact on my grids. I have not } have any trouble since I have been here. } I have changed everything, new staining plate, cleaned and scrubbed } everything that I use for staining. I still get some UA artifact. Is this } normal and should I just forget about it or is there something else wrong I } am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I } stain for usually 10-12 minutes. the grids before staining or clean, but } after UA stain they are a bit dirty... } } Any suggestions? } } Thanks, } } Eric A. Rosen } UCLA Medical Center } Dept. Pathology } EM Lab }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear all, We have a problem of saw toothe edges in JEOL 5410 SEm in scan 1 mode after x5000 Magnification.We tried changing OL,Cl aperture,Reducing filament spacer to 2.0mm.But no reults.The picture in slow scan is fine up to x20k.In higher Magnification we couldn't focus the image properely.(the sample used sputtered gold).Expecting everybody suggestion.Thanks in advance.
Regards,
A.Rajagopal Blue star Ltd Bangalore India. _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
I am in the process of figuring out a way of permanently mounting desmids (unicellular green algae with a cellulose cell wall) and was wondering about the relative benefits of DPX and Canada balsam as mounting media.
It appears that no-one in our field has done this kind of thing before (please tell me if I am wrong) and we have to guess our way through this.
Is Canada balsam still used? Which mountant has the better long term and optical characteristics in your experience? Has anyone got DPX slides which are older than say 20-30 years, and are they still acceptable?
many thanks
Micha Bayer
______________________________
Dr. Micha Bayer Royal Botanic Garden Edinburgh 20A Inverleith Row Edinburgh EH3 5LR Scotland, U.K. Tel. (+44) (0)131-248 2915 or 248 2965 Fax (+44) (0)131-248 2901 Project Homepage at http://www.rbge.org.uk/ADIAC/ RBGE home page at http://www.rbge.org.uk ______________________________
i am trying to find a way into the literature on experimental techniques appropriate to study 511 photon detection and capture vs properties of scintillating materials. Is there a standard way to do this?
Can someone point me at a good place to begin a literature survey?' sorry, my question is not better phrased, but I am just beginning to look at this?
Dear Sterling, It's been a while since I worked with these kinds of detector. You may be interested in either of two different types, depending on what the experiment is. Alkali halide scintillators are inexpensive and can be made large enough to cover a big solid angle (so two of them in coincidence can register the positron annihillation photons), but their energy resolution is rather poor. Germanium solid-state detectors are more expensive, but also can be made to cover a big solid angle to detect photon pairs, and their energy resolution is excellent. Find as up-to-date text on nuclear physics as you can, and check out the referrences from the chapter on detectors. Also check out the web for manufacturers of these detectors--Ortek and Canberra are the two biggest. Both companies have technical manuals which describe the detection process. They carry the solid state detectors and may also carry alkali halide detectors (or know who does). Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Here you have a summary on the information about IP systems for TEM. Thanks to everybody for the help!
Regards, Julian
Their linearity is over 5 orders of magnitude as opposed to 2 orders with film. However their resolution is much poorer. Something like 50 to 100 microns versus 2 microns with film.
bob j
I believe the pixel size is 30micron instead of film's 5u, so you cannot enlarge as much subsequently (or you need to use 6X more mag for same enlargement capability, and end up with 1/36 of area).
On a general way, the interrest of the IP is that its reponse in linear on a wider domaine than the Ag film. So you can enhance contrast if you have a small contrats in your sample (biology), or you can take a big dynamic in one exposure (for exemple in CBED mode).
The harware is expensive when you buy it, but the use is from no cost (apart the computer ; images are more than 20 Mb big !).
We use it, but we use also the film, and we will not stop the film soon.
But contact J. Werckmann. He can say much more about it than I.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
I did study this subject a few years ago and find that this technique is not well optimized for EM. For many reasons:
- cost of the system is similar to the EM digital camera - you require to load/unload Imaging Plates into the cassettes (you waste the time) - it takes from 2 to 5 min to scan one Image Plate (waste time) - I am not sure exactly, but seems to me, that you have to scan Image Plates soon after tacking picture, otherwise the image will deteriorated, and seems to me they are sensitive to light too. - you don't have a chance to adjust your image before recording (as it happens in case of digital cameras). -number of "shots" is limited and Image Plates are princely - Image Plates may be damaged during loading/unloading which may reduce declared number of use (a few thousand times I believe).
The good things about Image Plates are: - Image plate has more pixels than EM digital cameras, so the resolution is better. - you don't have to make any changes in the microscope. - the linearity and dynamic range is similar to the EM digital cameras - easy to use and less problem related to the equipment.
My conclusion was: technically, it's a great stuff, but it does not help in practice. You still need some help from technician to load/unload plates and scan them and you have to pay a 20-40$K for that beauty. Not so promising technology for TEM at least.
This information based on my "study" performed a few years ago and may not reflect modern improvements in this area.
Sergey.
All digital cameras including Gatans are approximately 10 times more sensitive than 4489 film. I know it for sure because just returned back from Gatan's demonstration. Image Plate should have very similar characteristics in terms of sensitivity, dynamic range and linearity as modern digital cameras. The best things about Image Plates it's their pixel size: 5000 x 4500 total pixels per plate. It's greater than in digital cameras: 1000x1000/2000 in most cases. It's still less than you could get from film with $800 scanner: 6400x4800 for 1600 optical dpi. BUT!!! You have LOAD/UNLOAD plates into cassettes manually. In the dark. Stack them and load into the scanner (so, you have to have space for the scanner in dark-room). Scanner will process them automatically (and at that time you could not use dark-room for regular things, light from scanner). Load them back into cassettes, degas magazine and load it into microscope. I don't remember exactly, but it seems to me the cost of one plate is something around $100. So, I have two magazines with 50 cassettes each: 100x100$ plus scanner plus salary for technician plus computer and storage media. Ok, I forgot to mention that you have to ERASE plates before use, also, using special eraser. I don't remember, how many plates you could ERASE in one run. May be it's automatic too. Scanning: 3.5 min per plate x 50= 3 h.
As I mentioned before, the technique is great, but not so helpful for regular day-to-day basis EM and does not save time so much. It works fine for molecular biology applications in "phospho-imagers" but people need to scan a few plates per day (even one a week) and they save a lot of time: 10-15 min exposure vs 5-10 days! I just don't see reason to switch to this technique to myself. If I'll have such money, I would buy digital camera. It's more practical solution from my point of view.
Sergey
There are advantages and disadvantages to all imaging systems. In some instances CCD cameras are preferable to film, in other instances film is superior (especially if wide field of view is a factor). What is best to use largely depends on the details of your experiment. I thought I would provide some information on imaging plate systems, and indicate one strong advantage the technology has over other recording media/systems.
Image plate systems use the "photostimulated luminescence" (PSL) phenomenon. Regions in PSL materials that have been previously exposed to a short wave length irradiation, will luminesce when stimulated by a second longer wavelength irradiation. The stored energy is stable until released by the second irradiation. Imaging plates are made using PSL films of barium fluorobromide (= 5 µm grain size), containing a trace amount of bivalent europium as a luminescence center, which are uniformly coated on a polyester support. Imaging plates replace conventional photoemulsion film in the transmission electron microscope and are exposed the same as normal film. To read the recorded image, the imaging plate is latter scanned by a red diode laser and the luminescence is collected through a light collection guide to the photo-multiplier tube (PMT). The analog current produced by the PMT is converted to a 8 to 16 bit depth digital signal. The pixel density can be read at between 50 to 400 pixels/cm. Imaging plates can be fully erased for reuse, but have a finite lifetime.
One main advantage of PSL films in their ultrahigh sensitivity (e.g. 2 x 10-14 - 2 x 10-9 C-cm-2 at 100 kV) to electron irradiation making them valuable for biological work (these are values quoted by a manufacturer Fuji). For example, film emulsions require 0.1 to 1 C-cm-2 to record an optical density S = 1 at 100 k magnification (see Reimer, 1975). Slow scan CCD cameras offer better sensitivity than film, however are inadequate for imaging certain biological specimens. Furthermore, CCDcameras have limited fields of view as compared to film. As an example, Sugi (1997) and coworkers have used a Fuji imaging plate system to record micrographs of myosin head movement induced by ATP hydrolysis in an environmental cell TEM. I avoided using the term “living myosin” since a biomolecule is not strictly alive, but it was biologically active. Electron doses low enough not to damage the delicate biomolecule could be used to record images on an imaging plate.
Reimer, L. (1975) in Physical Aspects of Electron Microscopy and Microbeam Analysis, eds B. M. Siegel, and D. R. Beaman, John Wiley & Sons Publishers, New York, New York, 231-245.
Sugi, H., Akimoto, T., Sutoh, K., Chaen, S., Oishi, N., and Suzuki, S. (1997) Proc. Natl. Acad. Sci. USA 94, 4378-4382. I contacted ditabis several months ago. Beyond a confirmation email from the company, I haven't received any from them.
Steve Widing Temple University
The Image plate system is awesome for materials applications as well.... !!
It is very easy to obtain diffraction patterns that include varying intensities (e.g., a spot pattern mixed with Kikuchi lines) with just one exposure on a single plate. The plates also show the slightest deviations in contrast which are not discernable using film, a CCD camera, or the TEM viewing screen. For example, differences in contrast in amorphous silicon oxide due to the multiple processing steps in integrated circuits may be easily observed with the plates. The advantage in the plates is evident in the low to intermediate magnification ranges where large variations in image contrast can be manipulated off-line. Each IP file is quite large (25MB) and therefore, contains plenty of information which can be manipulated before reducing its size that is suitable for publication purposes. The disadvantages to the IPs are the cost (although I believe the cost is dropping) and the extra processing time needed (~ 80 minutes for a batch of 32 plates).
The CCD camera is still very useful in the high resolution imaging regime where the intensity is rather uniform throughout the field of view and one can instantly observe the quality of the image. We will be evaluating high resolution CCD images with high resolution images obtained with the IPs in the near future. We are not sure what effect (if any) vibrations from the film mechanism will have on the high resolution images.
******************************************************************* Lucille A. Giannuzzi, Ph.D. Associate Professor, Mechanical, Materials, and Aerospace EngineeringDirector, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305Orlando, FL 32826email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321
We have just been testing the Ditabis IP scanner and were impressed.
It doesnt quite replace film or I think a good small-pixel below-screen CCD camera for resolution, but is much better than a whole field above screen CCD camera for resolution, and for dynamic range is in a class by itself.
We tested with diffraction patterns showing bright and dim diffuse features, convergent beam patterns, wedge-profile ion beam thinned sections, and normal and unstained biological material.
For low dose work, 1-2 seconds exposure at beam intensities barely visible to the eye was no problem. The resolution problem may be avoidable in practice because images may be taken at higher magnifications and lower beam intensities than normal for film - that is just a question of getting used to matching the imaging conditions to a different medium.
PLate handling is easy. The plates are loaded into the TEM magazine in the light. AFter electron exposure they are kept dark, but loaded into the scanner under light as dim as convenient - we used the light from the monitor screen.
Storing the plates for up to five days made only a very small difference to the intensity histogram - we didnt test beyond that. Plates can also be rescanned with very little loss.
The only real inconvienience we found was the impossibility of recording plate number, mag etc on the IP plate, as we are used to on the negative. If we had our own system we would mark the plates permanently in some way that showed on the readout.
IP plates are complementary to CCD cameras in some respects. You dont get the advantages of immediate viewing, but more dynamic range, more pixels, and of course one scanner can take plates from any number of TEMs. Finally, the price was the sticking point for us at this time. But if a cooperative lab somewhere else on this continent gets one we would likely buy a set of plates and work through the postal system until the price drops or somebody comes up with a "must have" application!
regards Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
Dear Roy, Even worse than that. It's almost trigonometry, and the great unknown, tube length! The best way to determine the mag of photos already done, for which you know the ocular and objective combination, is to go back to the scope with a specimen of known dimensions - I use something called a stage micrometer - a micro-ruler on a slide that has gradations down to 10um. I have photos (rulers) for every combination on my scope. I do the same for the CCD camera I sometimes put on my microscope in place of the film camera. My suggestion is to go down(over) to the department of Biology and see if someone can loan you a "Stage Micrometer".
Regards,
Fred Monson
} ---------- } From: Beavers, Roy } Sent: Friday, June 29, 2001 5:16 PM } To: 'Microscopy Listserver' } Subject: Microscope magnification } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Need some help trying to figure the magnification of some photos taken } with } the following: } } Nikon Optiphot-Pol Microscope } CF eyepiece 10X } CF objective 20X } } with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder } with a 4X marked on it. } } Focus for images was done through the monocular eyepiece on the camera } assembly. } } I assume that my magnification though the microscope eyepiece would be 30X } I assume that the magnification to the camera back would be 24X } } Sorry if this seems like a silly question but I do not do much optical } microscopy and got to thinking that it may be more than just the sum of } the } optics magnification. } } Thanks } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu } } }
} ---------- } From: Barbara Plowman } Sent: Friday, June 29, 2001 5:46 PM } To: fmonson-at-wcupa.edu } Subject: Re: RE: long-term storage OR what has happened? } } Excuse me, but might you need your medication? Enough... } } } } } "Monson, Frederick C." {fmonson-at-wcupa.edu} 06/29 7:30 AM } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } You are absolutely right, and most of the time we behave just as you } describe us. Patient, etc. One purpose of my tirade was to point out } that } there are differences between today and yesterday that give ALL of us } choices to make. I know a high school student who failed the year, } because } he wanted to punish his parents. When he told me what he had done, I } responded that I knew a young man just like him when I was in high school. } His solution was to get straight A's so he could become more quickly } independent. } Science is our own, very human, endeavor. Those of us who do } science must constantly work to remove the temptations to embellish our } results, to twist the statistics and to 'jump' to conclusions. But, and } here's where I differ perhaps slightly. Sometimes, there is a good reason } to yell! My wife hates my streak of sarcasm! But I was raised on the } stuff. When I did something 'dumb,' everyone who loved me, joined in the } derisive repair. I learned to think at my dinner table before I was 10. } I } did not like to be the but of everyone's attention. My Mom taught, then I } taught. I only stopped, because no one [too few] wanted to learn any more } [bad generalization]! Go to college today, and what do you find? Young } people unprepared for the rigor. Young people unprepared for the } intellectual dance - as we used to call it. Young people who don't know } what they want to do and are unwilling to work to find out. "They only do } what they are taught!" This is my mantra when I look for explanations in } the mirror. My older son had to learn - for the test - to name NH3 as } "ammonium", because his chemistry teacher would not believe the Handbook } of } Chemistry and Physics. My chemistry teacher in public high school took } chemistry courses at Rutgers every third summer. He was the expert in } chemistry. My physics teacher had a PhD in the subject. My biology } teacher } went on to be the chair of Physiology at a large Southern medical school. } If I wanted to complain about a professor at Lehigh, the Dean would show } me } a list of nearby schools that had openings. The attitude was clearly } communicated. The greatest lessons in life are those that teach one to } manage the worst parts of it. If I failed a course, it was MY failure - } and } it was at least MY problem! I wanted to be what I am, once I figured } everything out. The only advice I ever received, when I was having } problems } with learning, was, "Study harder." My Mom just kept on telling me to } keep } on trying as hard as I could. If I were young now, I'd be in group } therapy } and zapped on some pharmaceutical, if the system had its way. } Given my experience, I hope it is not difficult to understand why I } might not appear to understand the problems of others. BUT! If I see a } young person in science who is poorly informed about how it is done, then } I } will act in that person's behalf even if I am thought, and reported, to be } insensitive. } When I was a graduate student, we were all ordered to write } faculty/course evaluations. I did. I wrote analyses of each course, and } every one of them was critically positive. Not one word of my analyses } were } printed. If I had demonized one of my professors, I would have been } writing } novels today. The people who ordered the writings were ex-faculty } administrators who were pandering to the new consumer mentality in } education. If I had gone to my undergraduate department chair to complain } about a grade, he would have told me that I was wasting his time. Five } years later, there were lines of boomers outside of department offices. } The } generation of manipulators had arrived! While they were standing in line, } I } was learning. While I was learning, they were just getting warmed up! } I have had ideas snitched! Sometimes I even gave them away. On the } other hand, I have also had the other side of the experience. For six } months of learning followed by 30 minutes of consultation and work, I had } the good fortune to become a molecular biologist. For that effort, the } individual to whom I gave my best remembered my part - even after he had } spent years doing the work - and included me in the author list for a } gene(TERE1) that mapped to Chromosome 1 and appeared in print in a Nature } publication. He did NOT have to do that. There was NO obligation nor } agreement. I helped because he asked. People are basically good, } especially scientists, and I always start with TRUST, and I always will. } You are still absolutely correct. } } Regards for YOUR sensitivity, } } Fred Monson } } } ---------- } } From: Sousan Abolhassani } } Sent: Thursday, June 28, 2001 8:07 AM } } To: Paul Webster } } Cc: MSA listserver submission } } Subject: Re: long-term storage OR what has happened? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Paul, } } } } The point about this server is that it is used by } } a very heterogeneous public from which a non-exclusive } } list is the following: } } } } students, } } professors, } } scientists/ } } researchers } } technical staffs } } suppliers } } producers } } specialists } } non-specialists } } etc. } } etc. } } etc. } } } } and all these from so many different backgrounds with so } } many different problems and even from so many different countries... } } That if we want to teach all of them to behave as we expect } } before they post a query, the server could go to early retirement. } } } } I guess the philosophical attitude that one should have is to } } really try and understand both sides of the discussion. } } } } 1- on one hand the trust that professor Monson is demanding } } is not there because our users are a small sample of our } } society. How many times have we heard that people have had } } their ideas picked by others and their publications stolen } } by others and .... in the very scientific community that most } } of us hoped to be the purest part of the society. Still } } the scientists are generally speaking very sincere people. } } So if someone is not having trust to others, it's the } } consequence of what they have observed in their environment. } } } } 2- I do not know if Rachel is a student (may be not), and if she } } really was not trusting or just did not think to write the reason and } } so on, so we do not discuss the case on a specific way. } } But talking about going to the library, there are labs } } where the students have really not access to the books and there } } are places where superiors do not give the support the students } } and other researchers' need, or simply there are not } } enough books available. } } } } So there are always reasons for the way people behave } } and the more our society goes towards a very materialistic and } } "win whatever the situation" attitude, the more the arguments } } of professor Manson will be applicable because the more people } } will behave in the way he is not happy about, and I understand } } and apreciate his point. } } } } All that apart, it is better to listen to people,as often things } } that are not to be done are not published and are only "known". } } Of course provided that answers are given generously, as arrogance } } for giving is so destructive, even for donator. } } I therefore agree with the fact that we should just be able to } } ask the question and someone will react to give us an answer. } } } } I think that professor Monson has evoked a point that is of } } great importance and that is the human aspect of all we do, as } } "human" is what we are hoping to be at a first place. } } That needs sensitivity... and continuous effort. } } } } Regards, } } } } Sousan } } } } } } Paul Webster wrote: } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } Dear Colleagues, } } } } } } The messages from Professor Monson in reply to a question about } } long-term } } } tissue storage makes me wonder what has happened to this listserver? } } } } } } His message was highly entertaining and even if the poster of the } } original } } } message may have felt "picked on", there was no reason to expect him } to } } post } } } an apology, no matter how informative it was. } } } } } } Far too many posters use this listserver as a way of finding a quick } fix } } to } } } simple solutions to technical problems that can be easily researched } in } } text } } } books. } } } } } } Often these messages contain little background information on the } } technical } } } problem under investigation. Even so, many of these questions get } full } } } attention and detailed replies, even if the replies are not posted on } } the } } } main list for public view. } } } } } } What has happened to the free and open discussion of issues and } } protocols } } } that we used to have? Are people so afraid of being accused of being } } } incorrect that they do not post their replies openly? Or maybe people } } are } } } afraid of being accused of being impolite. } } } } } } I look forward to controversy and hope that my comments are read } } carefully } } } and replied to with vigor. Only in this way will I be able to learn } new } } } thing, and this is the main reason I still subscribe to this } listserver. } } } } } } If I am factually wrong about something, tell me. It is better to } find } } this } } } out on the listserver than in a more professional (or cruel) } } environment. } } } } } } As for discussion subjects: why was there no comment about the } } unadvisable } } } suggestion made about storing samples in a frozen state? Would anyone } } } working with EM really choose to store their samples at -20 degrees } } before } } } processing for TEM? } } } } } } Where is all the discussion about extraction that can occur when } storing } } } samples in alcohols or buffers, or on the contamination that I know } } grows in } } } everyones old cacodylate buffers? } } } } } } Do we not comment because we know it has all been published in } } textbooks? } } } } } } Dear Profesor Monson, do not feel bad about improving our general } } knowledge } } } and our professional etiquette. I for one support your views and find } } your } } } comments useful and entertaining. Please keep up your submissions. } } } } } } Best regards, } } } } } } Paul Webster } } } } } } Paul Webster, Ph.D. } } } Scientist II and Director } } } Ahmanson Center for Advanced EM & Imaging } } } House Ear Institute } } } 2100 West 3rd Street } } } Los Angeles } } } CA 90057 } } } } } } pwebster-at-hei.org } } } p: 213 273 8026 } } } f: 213 413 6739 } } } http://www.hei.org/research/depts/aemi/aemi.htm } } } } } }
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Eric, A couple things to check: (1) 12 min staining may be longer than necessary....try 5 min and see what happens.
(2) What about the water used for rinsing? This can be a source of contamination. Always use double distilled water. I used to use the bottled water used for reconstituting vaccines when I worked in a medical center and had easy access to it. At the time I had a less than reliable water source.
(3) Try washing by moving your grids through a series of water droplets prior to "jet" washing with a squirt bottle. This eliminates retension of stain along grid wires.
(4) Are you sure you are not getting contamination from phosphate percipitation (you didn't say what fixative you used for your material). If the sample has a high calcium content and is fixed with a phosphate buffer, it can lead to percipitation of a fine electron-dense compound....especially if the tissue also has high lipid concentration.
I would run some sections from blocks containing other tissues that have worked well in the past as a method control. If they are clean and your problem sample is not, then look for the dirt source prior to the staining step. Good luck...these problems are a never ending source of frustration. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
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Hi there fellow microscopists,
Here in the Pathology EM lab we do not do rocket science, but standard thin-section TEM of renal biopsies.
In the last two weeks though I have been getting some ka ka (technical term) on my grids from my UA stain. I have already made up two new batches of UA stain and use .45 micron filters just before use. Question is that all of the sudden I am getting UA stain artifact on my grids. I have not have any trouble since I have been here. I have changed everything, new staining plate, cleaned and scrubbed everything that I use for staining. I still get some UA artifact. Is this normal and should I just forget about it or is there something else wrong I am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I stain for usually 10-12 minutes. the grids before staining or clean, but after UA stain they are a bit dirty...
Any suggestions?
Thanks,
Eric A. Rosen UCLA Medical Center Dept. Pathology EM Lab
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Roy, Invest in a good micrometer slide and use it to determine magnification. It may seem expensive initially but, since it can be used on any coumpound microscope regardless of set-up and camera, it will pay for itself in convenience (and accuracy of determining magnifications) for years to come. You can purchase them through your microscope venders for both compound and stereo microscopes. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
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Need some help trying to figure the magnification of some photos taken with the following:
with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder with a 4X marked on it.
Focus for images was done through the monocular eyepiece on the camera assembly.
I assume that my magnification though the microscope eyepiece would be 30X I assume that the magnification to the camera back would be 24X
Sorry if this seems like a silly question but I do not do much optical microscopy and got to thinking that it may be more than just the sum of the optics magnification.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
At 4:16 PM -0500 6/29/01, Beavers, Roy wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I have cross sectioned this sample along the axis of the wires and see interesting features.
The defect terminal was determined to contain 13% Cr (EDS standardless analysis of the Cross Section), whereas the other materials were determined to contain 21% Cr. I checked with my client and he had chosen to use a lower grade material for the defect terminal, because it had the same high temperature ratings. I was able to peel back the cross sectioned element (21%Cr material) and examine its surface as well as the surface of the hole wall (13%Cr terminal material). The hole wall was completely covered by a binary particulate material. The particles contained either Ti and oxygen or Al and oxygen. The element material was covered approximately 70% with this material. Examination of the cross section showed this layer, but also a particle field that was evenly dispersed through the defect terminal and ended a few hundred microns past the weld interface between the terminal and the element. These particles have a lower Z than the matrix and are on the order of 1 micron. Elemental analysis of the particles embedded in the terminal material shows mostly Ti, C, and N, with lower levels of Zr.
My proposed mechanism for the failure is that either the media in the furnace or the refractory material of the furnace has corroded the surface of both the "A-1" grade element and the lower grade terminal material. The lower grade terminal material has also had a migration of the surface corrosion into it internal structure and into the vicinity of the weld. This internal corrosion is what eventually gives rise to the electrical failure. The flowed material, that was initially investigated, I believe to be a "symptom rather than a cause of the failure (as Woody White had stated)".
My questions now are: Is this a plausible mechanism?
Can the corrosive particulate, on the order of 1 micron, migrate 100's of microns into a low grade ferrous alloy at temperatures of 1100°C? (Although localized temperature could reach higher levels)
I have images if anybody would like to see the features I discuss.
-----Original Message----- } From: Smartech [mailto:smartech-at-optonline.net] Sent: Wednesday, June 27, 2001 1:33 PM To: To all on the list
I'm curious about whether there's any published data on the average lifespan of scanning electron microscopes in an industrial environment. Our first SEM we had for 15 years, now we're up to 13 on the current system. My manager, being the sort of histogram-oriented guy he is, asked me for some longevity data. In the absence of any previously published data, I'd sure be interested in individual experiences along these lines.
Much obliged,
Dave Stadden Research Scientist, Testing and Analysis Lab Armstrong World Industries, Inc.
Dear Eric, You said nothing about the solvent.??!!!? While switching out a filter on the building supply, did someone make an error? Etc.
You didn't mention the morphology of the ka ka. Smaller than .45um?
Are these the clumps of homogeneously very small crystals/grains/? that often appear in older mixes/solutions?
Has someone else taken sample from the bottle recently?
Do you do your staining on an open bench? In a hood? In a Petrie dish?
Try .22um filter. If still present, then how old is jar of U salt?
Some other things to think about:
Biological processes involved with U depletion/removal in environment may have applicability to lab situations. See: (http://www.nttc.edu/env/Uranium/Uranium_chap8.html). Almost ALL UAc(Uranyl acetate) has some trace of Fe in it - increasing the possibility of a slowly developing linked biological process leading to production of "increasing" amounts of UO (fine grain ppt?) in the reservoir of UAc. DID you know that uranium (U) is 'pyrophoric'? Then there is all that acetate! See: (http://www.geol.vt.edu/research/gssrs/gssrs2000/abstracts/tkendall.html).
What's different about the specimen, or is this happening with ALL specimens?
[With regard to your concern about your concern. No, you should never become complacent. Such aggravations must be removed. You must cultivate your crisis management skills. If you don't, you will feel guilty and inadequate. It would be better to manifest OCD than to feel 'guilt and 'inadequacy.']
Regards and best wishes,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Eric } Sent: Friday, June 29, 2001 6:24 PM } To: 'Microscopy Listserver' } Subject: UA ka ka on grids } Importance: High } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there fellow microscopists, } } Here in the Pathology EM lab we do not do rocket science, but standard } thin-section TEM of renal biopsies. } } In the last two weeks though I have been getting some ka ka (technical } term) on my grids from my UA stain. I have already made up two new } batches } of UA stain and use .45 micron filters just before use. Question is that } all of the sudden I am getting UA stain artifact on my grids. I have not } have any trouble since I have been here. } I have changed everything, new staining plate, cleaned and scrubbed } everything that I use for staining. I still get some UA artifact. Is } this } normal and should I just forget about it or is there something else wrong } I } am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I } stain for usually 10-12 minutes. the grids before staining or clean, but } after UA stain they are a bit dirty... } } Any suggestions? } } Thanks, } } Eric A. Rosen } UCLA Medical Center } Dept. Pathology } EM Lab } }
"Hypericin" - the active principal in St. John's Wort, a quencher? See: http://www.phys.uni.torun.pl/~lum98/p_abstr/hovhanis.html
It apparently absorbs strongly in the yellow!! See: (http://www.aspjournal.com/public/vol71/iss2/abstracts/abstractvol71iss2pp18 8-194.htm)
Might be worth a look.
In any case, the principle of quenching autofluorescence should be clear to those wondering what's going on.
Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Shea Miller } Sent: Friday, June 29, 2001 11:51 AM } To: microscopy-at-sparc5.microscopy.com } Subject: counterstaining for immunofluorescence } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi again; } we are having a wee bit of trouble distinguishing our signal from the } autofluorescence in our immunoassay on immature soybean seeds/seed coats. } So far, we have used a light stain of toluidine blue, but I'm not } completely happy, and am considering Evans Blue instead. What do others } use to quench the yellow autofluorescence (our Ab is FITC labelled) in } plant tissues, and when do you actually apply the counterstain? Before } the Ab incubations, or after?? } } thanks so much again in advance } shea } } } } Dr. S. Shea Miller } Agriculture & Agri-food Canada } Eastern Cereal and Oilseed Research Centre } Central Experimental Farm } Ottawa, Ontario } Canada K1A 0C6 } Phone: (613) 759-1760 } Fax: (613) 759-1701 } Email: millers-at-em.agr.ca } } }
While I cannot remember ever seeing migration of particulate oxide into a metallic, it is certainly possible for oxygen to diffuse into the material and (then) form oxides.
I have not checked references for the materials in use, but typically, the alloys with lower Cr concentrations are more likely to oxidize/corrode. Again, I need to check the references to be exact, but chromium oxide has a fairly low vapor pressure at 1100 C. Not only can diffusion occur at this temperature, but you may also vaporize cr that has oxidized and re-deposit nearby on cooler surfaces.
As a matter of fact, the first pix I saw of the oxide crystals looked more like a vapor deposition structure than a product of surface corrosion.
Another thought... As the electrical joint oxidizes and begins to fail, the resistance likely rises. For the same power input, this would result in localized heating which could go far higher than the 1100 C process temperature. Higher temps would equal faster oxidation / diffusion, further raising the resistance. The worse it gets, the worse it gets...
Hard to accurately evaluate all variables long distance, but it sounds like you are making progress and narrowing the possibilities. I am still concerned about any lube residue which could break down at temp and liberate elements (hydrogen, oxygen, halogens, sulfur, etc.) which could accelerate corrosion for many materials.
I would like to thank all of you who have responded to the solicitation for suggestions for symposia for the M&M annual meeting in 2002 in Quebec. The response was very strong and it will be plain impossible to accommodate all suggestions. I thought it worthwhile to mention that the biological sciences area has received far more suggestions than the physical sciences area. Is that a sign?
Anyway, I want to very much thank all of you for the suggestions and at the same time ask for your forgiveness that I will not be able to respond properly to all who have put up the effort to write. If you will not hear from me over the next few weeks, please be assured that your suggestions have at least contributed to emphasize certain directions for the symposia.
It is great to see how active our society is.
Best regards,
Edgar Voelkl Program Chair M&M 2002
--
___________________________
Dr. Edgar Voelkl Program Chair M&M 2002
Oak Ridge National Laboratory P.O. Box 2008 Bldg 4515 Oak Ridge, TN 37830-6064
I have a Zeiss EM-9S-2 TEM for sale. Needs vacuum pump and chiller. Unit is in excellent condition. Have all manuals and many miscellaneous spare parts. Best offer. Buyer responsible for packaging and freight.
Contact:
Bradley K. Starcevich microscopist-at-excite.com
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
At 3:24 PM -0700 6/29/01, Eric wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
****************** I know this sounds like a cop-out (am I dating myself with that phrase, or what?), but try a new bottle of UA. I had the same problem years ago, and that's what I finally resorted to. It was easier and more comfortable than continuing to knock my head against the wall! Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I think the issue is probably determined by when support gets hard to find or when the accumulation of new features reaches a critical level.
We have a JEOL 840A that is about 15 years old and is still doing fine. It does not offer the integrated control interface to our EDS system like newer units might. But the imaging is still good and JEOL is still supporting it. I can easily imagine it going another 5 years or more.
I guess I would toss out 15-20 years as an expected life. Labs that insist on having the latest and greatest might turn a scope around in less than ten years.
That's one man's opinion (whose newest car happens to be a 1989). Warren
At 11:46 AM 7/2/2001 -0400, you wrote:
} I'm curious about whether there's any published data on the average } lifespan of } scanning electron microscopes in an industrial environment. Our first SEM we } had for 15 years, now we're up to 13 on the current system. My manager, being } the sort of histogram-oriented guy he is, asked me for some longevity } data. In } the absence of any previously published data, I'd sure be interested in } individual experiences along these lines. } } Much obliged, } } Dave Stadden } Research Scientist, Testing and Analysis Lab } Armstrong World Industries, Inc.
In a recent discussion of how to avoid algal growth in water chillers, someone suggested using a rather high concentration of ethylene glycol. I have serious reservations about employing this method. Ethylene glycol is a hygroscopic substance, and fairly high concentrations of it are required to be effective. If for any reason (and water lines have been known to leak on numerous occasions and for a variety of reasons) some of this solution gets into any part of an instrument (and I have seen cooling water get onto circuit boards and into various internal parts of instruments on several occasions), it can be very difficult to remove. Worst of all, because of its hygroscopic character, any residue that is left can pick up water from the atmosphere, keeping that part of the instrument moist forever after. The control of algal growth in electron microscopes is discussed in more detail on page 216 of my book, 'Vacuum Methods in Electron Microscopy (for a description see http://2spi.com/catalog/books/book48.html and http://pup.princeton.edu/titles/6484.html) -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Dear Jon, We at the University of Kentucky also have a core facility that originally was just EM. We have also experienced a slow down in TEM use compared to years past. In the last three years we have added 2 confocals. We have an upright and an inverted, both Leicas. Most of our users are not in our building but no one seems to complain about the hike. They just cover their samples with foil and make the trek. We expect everyone to supply their own antibodies and we supply training, help and advice (for a price of course). We have sign-up sheets and on busy days limit users to 2 hour blocks of time. Having two instruments does help if someone needs to do a time lapse study. Users constantly have questions or problems that need to be resolved, so it's important that someone in the lab be quite familiar with the instrument and it's software. For the users who prove to be proficient, we allow after hours use. Otherwise someone is always around . The group whose money purchased one of the instruments pays the same as everyone else. The biggest problem we've had is making people wait when the instruments are down. Service has been very slow. Let me know if we can help you in any way and good luck.
Mary Gail Engle, manager Electron Microscopy & Imaging Facility University of Kentucky Lexington, KY
I have a Zeiss OPMI surgical microscope. I bought it used as-is. It came with a bulb that didn't last. The filament broke away from the support wires. A fine spiral of thinner wire wound around the support wires seemingly to help support the filament, and it had spiralled off the supports.
Fortunately my junk pile revealed a replacement bulb. However, after the 'scope took a trip in the car to demo for some kindergarteners, its filament also broke and the thin wires spiralled away.
$45 at Bulbdirect.com and I'm back to a working bulb and a spare. What should I be doing to protect myself against early lamp failure? Cool-down without movement for 20 minutes? Remove lamps before driving around with the 'scope? Are short bursts of usage (5-10 minutes each) to blame?
- John
P.S. Dr. Monson: Love the stories. Keep in mind that all seemingly newbies on the 'net might be learning very slowly over time, just for fun, and without benefit of a degree program. The Web is our Oracle, but I'll try to pay attention during class. Please do not hit me with eraser, chalk or ruler.
What a coincidence! The same question came up here and I entered "511 photon" and asked "Google.COM".
I immediately received for example: http://cfi.lbl.gov/instrumentation/Pubs/4on1.pdf
That seemed to satisfy my questioner, so I quit. I hope someone who really knows the field is listening.
Fred Monson
} ---------- } From: sterling stoudenmire } Sent: Saturday, June 30, 2001 11:26 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: scintillation crystals } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } i am trying to find a way into the literature on experimental techniques } appropriate to study 511 photon detection and capture vs properties of } scintillating materials. Is there a standard way to do this? } } Can someone point me at a good place to begin a literature survey?' } sorry, my question is not better phrased, but I am just beginning to look } at this? } } sterling } } } Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find } the cure for cancer and other diseases. There will always be a need for } the trained clinician (MD/RN) but, advanced diagnostic and treatment } option } selection has become gene based, has moved from the physician's practice } to } the computerized cell and molecular biology laboratory, and appropriate } treatment options should now be based on the personal biology of the } patient. } }
Dave, It really depends upon how the system is cared for. I'm still servicing microscopes that date back to 71 that work just fine. They were 6 years old when I STARTED servicing them. Red Lion High School (just across the river from you) has an SEM that's older than at least one of the teachers who uses it (and all of the students).
A more germain question would be "Is the current microscope able to do the things we require, and do them well and efficiently?" If the answer to this is "yes" and you have adequate service to keep the system running, there is probably no reason to even think about replacing it.
On the other hand, if newer technologies could improve your performance significantly, then you might want to consider upgrading. The downside? When the PC that is running the latest system becomes obsolete (3 years? 5 years?) and the rest of your system (vacuum, High Voltage, etc.) is still fine, what are you going to do? Try to find a new 486 with Win 3.1 to replace a crashed but fairly recent Amray, for example. Their boards and software won't run on anything newer. I'm not picking on them, I'm just familiar with them.
Alternatively, EDS systems have a notoriously short lifespan and it's getting shorter. Can the addition of a beam control interface and a new EDS system give you all the new stuff you need (or want)? This tends to be less expensive and easier to justify because of the historic turnover rate. The bonus being that all users are at least familiar with the basic SEM and don't have to start from ground zero (if you have a lot of users, this could be a consideration). This is an easy way to get digital imaging, for example.
Just my $.02. You should get a lot of interesting and useful replies.
Ken Converse owner Quality Images third party SEM service Delta, PA (York County)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } I'm curious about whether there's any published data on the average lifespan of } scanning electron microscopes in an industrial environment. Our first SEM we } had for 15 years, now we're up to 13 on the current system. My manager, being } the sort of histogram-oriented guy he is, asked me for some longevity data. In } the absence of any previously published data, I'd sure be interested in } individual experiences along these lines. } } Much obliged, } } Dave Stadden } Research Scientist, Testing and Analysis Lab } Armstrong World Industries, Inc. } } } } }
At 11:02 AM 7/2/01 -0700, Markus F. Meyenhofer wrote: } I am using a 50% Methanol and 50% ddH20 as my solvent for the solution of } UA stain. } } the particles are not fine grain, bit somewhat larger in size.. I have not } measured them as of yet, but will try to do that today... } } The solutions are relatively fresh.. I just made them last Monday... } } No one else uses this solution.. I mix it for myself to use... } } Staining is done on a clean petri dish with a dental wax base.. and } covered.. the grids are submerged in the droplet for 12 minutes.. } I had been doing 15 minute staining till I mixed up this batch of stain and } had to cut back on the time... } } I have on order some .2um filters.. } } 5 minute staining is too light.. and still has some schmutz on the grid... } } We have been using for the past two years a sterile water we buy in 1500ml } bottles. and have never had a problem till recently... } } I use a small set of three beakers that are filled with water and the grids } are dunked in each beaker 10 times then put on a clean piece of filter paper } ... } } We, use a modified karnofsky fixative... no phosphate
The lifetime for a given SEM depends greatly on the support from the manufacturer. As Warren has pointed out, the JEOL 840A has a life expectancy of at least another 5 to 10 years. JEOL has a policy of supporting their equipment in excess of twenty years. They use older machines that have been "traded-in" to support others when parts are unavailable from the factory. I can still remember having parts available for the JEOL U3 which is a tube model.
On the other side of the coin Amray & Topcon have since not supported the older SEMs and the SEM value and longevity has diminished. At present I have an Amray 2030C that I am considering using for parts. It is ashamed to do this as the instrument in only six years old & costs over 350K new.
My car is a 1992 Honda.
Regards,
Earl
----- Original Message ----- } From: "Warren E Straszheim" {wesaia-at-iastate.edu} To: {Microscopy-at-sparc5.microscopy.com} Cc: {David_R_Stadden-at-armstrong.com} Sent: Monday, July 02, 2001 10:42 AM
} From: Microscopy Society of America: Marketing and Communications Committee (MSAMCC)
Colleagues:
MSA is a vibrant society which encompasses all facets of Microscopy and Microanalysis. The society and it's members are dedicated to the promotion and advancement of the knowledge of the science and practice of all microscopical imaging, analysis and diffraction techniques useful for elucidating the ultrastructure and function of materials in diverse areas of biological, materials, medical and physical sciences. To this end MSA is a resource which is second to none throughout the world. If you're not currently a member of MSA, then you're missing out on both the opportunities and the fun; and we would like to take this opportunity to invite you to join our family.
If you are already an MSA member, take a moment to talk to your colleagues and/or students about MSA's great benefits and encourage them to join the society. Invite them to be come part of our world wide microscopy community.
MSA Membership Benefits Include:
* A subscription to the high-quality journal: Microscopy & Microanalysis -included with your membership dues http://www.msa.microscopy.com/MM/MscopyManalysis.html * Receive advanced information and reduced fees for the Annual Meeting - the preeminent microscopy related meeting and exhibition http://www.microscopy.com/MSAMeetings/MMMeeting.html * Access to extensive educational materials & activities http://www.msa.microscopy.com/RefEdu.html * On-line Directory of MSA members http://www.msa.microscopy.com/MSAMembers/SearchMembersDB.html * Opportunities to have your accomplishments recognized as an Awards recipient. http://www.msa.microscopy.com/MSADocs/MSAAwards.html * MSA Placement Office http://www.msa.microscopy.com/PlacementOffice/JobListings.html * MSA Certification Program http://www.cvmbs.colostate.edu/emcenter/msa/certboard/ * MSA Technologist Forum http://www.cvmbs.colostate.edu/emcenter/msa/techforum/ * MSA Public Policy Committee -remain aware and influence governmental funding opportunities http://www.msa.microscopy.com/PublicPolicy/ * MSA Sustaining Members Program http://www.msa.microscopy.com/cgi-bin/SMDBListing.pl * MSA Vendor News and Surplus Equipment Listings http://www.msa.microscopy.com/News/NewsListings.html http://www.msa.microscopy.com/SurplusEquipment/SurplusListings.html * Reduced prices for books and journals * Subscription to the MSA Bulletin to keep abreast of Society activities * Specialized networking via participation in "Focussed Interest Groups" with microscopist's having similar interests to yours * and of course the International Networking facilitated in part by the Microscopy Listserver to keep abreast of worldwide microscopy activities and information.
So if your not a member of MSA, take a moment and join! It is easy and inexpensive [$15/Students, $ 45/Regular, $63/outside of North America and don't forget this includes the a subscription to the Journal].
You can become a Member On-Line at http://www.msa.microscopy.com/MSADocs/MSANewForm.html or request that an printed application form be sent to you by sending an Email to: MSABusinessOffice-at-msa.microscopy.com
Remember MSA needs YOU ! --------------------
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Agreed, a stage micrometer is the best way, but they are not that expensive. As always, shop around, but I expect that you will find the microscopy supply businesses offer the better deal. We list five such micrometers and all but one cost less than US$150 including freight. Disclaimer: ProSciTech sell stage micrometers and so has an obvious interest Cheers Jim Darley ProSciTech
On Tuesday, July 03, 2001 1:10 AM, Debby Sherman [SMTP:dsherman-at-purdue.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Roy, } Invest in a good micrometer slide and use it to determine magnification. It } may seem expensive initially but, since it can be used on any coumpound } microscope regardless of set-up and camera, it will pay for itself in } convenience (and accuracy of determining magnifications) for years to come. } You can purchase them through your microscope venders for both compound and } stereo microscopes. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } } S-099e Whistler Building } West Lafayette, IN 47907 } } -------------------------------------- } Date: Friday, June 29, 2001 4:16 PM } } From: Beavers, Roy {rbeavers-at-post.cis.smu.edu} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Need some help trying to figure the magnification of some photos taken with } the following: } } Nikon Optiphot-Pol Microscope } CF eyepiece 10X } CF objective 20X } } with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder } with a 4X marked on it. } } Focus for images was done through the monocular eyepiece on the camera } assembly. } } I assume that my magnification though the microscope eyepiece would be 30X } I assume that the magnification to the camera back would be 24X } } Sorry if this seems like a silly question but I do not do much optical } microscopy and got to thinking that it may be more than just the sum of the } optics magnification. } } Thanks } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu } }
Somebody just asked me the reference for toluidine blue/sodium borate (1% each) which is our routine LM stain - I cant find an attribution in any of about 10 books. Can anybody help? This person should not be forgotten! One suggestion was Alsop?
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
In an old (1976) book on botanical microtechnique, there is a reference for 1% tol blue in 1% borax - Trump BF, Smuckler EA, Benditt EP (1961) A method for staining epoxy sections for light microscopy. J Ultrastr Res 5:343-348.
There's also Mallory's borax methylene blue, maybe tol blue was a substitute for the methylene blue?
cheers, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
SEM service life is based on parts availability and support.
Many older SEMs are still supported because the failing parts/components are still manufactured. Fortunately manufacturers used many readily available parts. Also many institutions/businesses have purchased new equipment and "retired the old". Fortunately there are some enterprising individuals who recognized a niche and began collecting "abandoned" SEMs. These individuals now provide a necessary parts/service to SEM industry.
As for support....if you can do much of your own you are smart. For those of us who neither have the time or expertise we need to seek out the experts/entrepreneurs.
I currently manage three SEMs. The oldest is AMRAY1600 purchased in 1984. It is a workhorse for us. Despite minimal maintenance and no service contracts over the past years it has performed well. Recently a number of components began to fail. It was suggested that the system be retired and replaced.
I found expertise in SEMTech Solutions Inc. During the past year we have replaced several aging components, added digital imaging, and moved the SEM to another building for access to multiple users. EDS upgrade is also being considered. As long as parts are available I expect the system to perform another 20 years!
SEMTech provided expertise and parts. I would highly recommend them for AMRAY instrument service.
c/o Jim Peterson PO Box 2155 Natick MA 01760 tel: (978) 663-9822
I have no vested interest in the company. I am simply a truly satisfied customer with an excellent service provider.
Finding other experts for other SEM instrumentation may be difficult. However, if the SEM community needs/supports it, I think there are individuals prepared to provide it.
Perhaps the reason these smaller new EDS ventures have been able to establish themselves is because the larger established businesses no longer listened/responded to their customer needs. A similar circumstance could evolve with SEM maintenance.
The question poses another thought: How could the SEM community "inventory" all SEMs by brand, model, age, location, condition, and status. Thus all SEMs being accounted for could effectively provide the potential inventory parts list and availability.
I have been seeking a double-tilt holder for our 2000fx scope for some time with no success. This holder is to be used for general teaching in the up coming semester. Purchasing a new holder takes a bit too long to be delivered. Also thought 2000fx holders are compatible with those of JEOL100 type scopes. So I may have a better chance to acquire a used one. Anyway, if you have one sitting in your lab with no immediate use, please contact me off line. Thanks much. Have a great holiday!
********************************* Chaoying Ni Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
As the image is in a SEM is created by lines of information, many problems exhibit themselves by giving saw tooth edges to the lines. The slower you scan the better the instrument is at synchronizing out the interference.
The most common reasons for the above are:-
1. Vibration 2. External magnetic fields
If we consider operating parameters we may be able to track this down. Magnetic fields will have a greater affect at lower accelerating voltages and at longer working distances, in both cases the final lens strength is reduced allowing the external field to overcome what protection the lens field provides. Try working at the shortest possible working distance and at the highest kV, probably about 5mm at 30kV. If this reduces or removes the problem it would be an external field that is the culprit. Another indication of an external field problem is the TV rate scan will have a sub ripple that will work its way down the screen.
We need to know if you have been able to take good photographs above 5,000X in the past. If so we then can assume the problem is a result of a change in the laboratory; a new piece of equipment is giving off the field. This may be many feet away, the field effect is according to the inverse square law. Try going round switching off new equipment and see if that helps. I have seen problems due to incorrect wiring of new equipment so if you find a field source have a technician check the wiring before going any further.
Please come back if you need more help.
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
I would like to perform a series of immunostains on cultured neurons to detect membrane proteins using confocal microscopy. The animal used is mouse and the antibodies were generated in rabbit. I plan to use a biotinylated secondary antibody and an avidin/streptavidin conjugated fluorochome that can be excited by the argon (488 nm) or HeNe (543 nm) laser lines of a confocal microscope. I need information on the source of these fluorescence compounds and your experience with Alexa/Cy dyes for this type of work.
Thank´s for your help.
Félix Viana
============================================================= Dr. Felix Viana Instituto de Neurociencias Universidad Miguel Hernandez / CSIC Apartado 18 San Juan de Alicante 03550 Alicante (SPAIN)
We are doing an IEM for SEM with bacteria and would appreciate any comments. We are looking for surface label 5nm gold with a FIESEM. Our protocol is:
3% PF 0.05% GA fix PO4 buffer
rinse
Bock in 1%NGS
rinse
Primary ABY incubation O/N cold
rinse
Secondary gold conjucated ABY 1hr
rinse
Hard fix 2% GA same buffer
rinse
2% UA filtered DH2O
Dehydration
Critical Point Drying
Our questions are is the UA enough to stablize the lipds throughout the dehydration and cpd? We believe osmium would strip off the gold and tannic acid may make small precipitates similar in size to the gold. We will use in-lens and backscatter to view the gold. Any other suggesstions would be appreciated.
The position described below was recently announced at the University of Colorado. Those interested in applying should contact Dr. Andrew Staehelin directly.
Tom Giddings
Thomas H. Giddings Tel: (303) 492-8402 MCDB Microscopy Facility Fax: (303) 492-7744 University of Colorado giddings-at-spot.colorado.edu Boulder, CO 80309-0347
} From: Andrew Staehelin {staeheli-at-spot.Colorado.EDU}
OPENING FOR PROFESSIONAL RESEARCH ASSISTANT
I have a job opening for a Professional Research Assistant starting sometime in August.
The research is on structure function relationships of the Golgi apparatus in yeast (Pichia pastoris) and plant cells. The work will involve cell culturing, the growing of Arabidopsis plants, light microscopy studies of cells expressing GFP constructs, electron microscopy of high pressure frozen cells, EM tomography and immunolabeling experiments. The successful candidate will also be involved in some lab administration duties.
Starting salary is in the $24,000 to $27,000 range, depending on experience.
Please send application, including curriculum vitae and names of three references, to Andrew Staehelin, MCD Biology, UCB 347, University of Colorado, Boulder, CO 80309-0347, or email to: staeheli-at-spot.colorado.edu. Application deadline: July 15, or until position is filled.
Andrew Staehelin MCD Biology University of Colorado Boulder, CO 80309-0347 U.S.A.
Gee, I wonder how many of us wear the same, multiple hats? I have been the director of the EM facility here for 13 years, and for the past 4 have also managed the optical microscopy core which consists of a confocal and a widefield fluorescence. We hope to add a second confocal in the next year or so. Our users come from various points in the med. school complex and we have some that come from other institutions. Travelling with prepared, fixed samples is not a big deal. Its only the folks who need to look at live cells that have a problem. Our confocal was purchased with an NIH SIG, but the main PI on that grant, who is also the Chair of the dept. in which it resides, pays the same fees as everyone else. We limit each user to 2 blocks of up to 3 hours each per week. People who have been fully trained and "certified" by me are allowed access 24/7. (Our fees drop to half after hours to encourage them!) If you have the space in your lab, go for it. My EM facility is in one dept. at one end of the complex & the optical 'scopes are in another dept at the opposite end. I can log a mile a day just walking between the 2 facilities on days when the confocal or its users are acting up.
Our users are responsible for fully prepping their own samples. We supply immersion oil & lens paper. As a courtesy, we stock CD-Rs and bill for what they cost us. In additon to the microsocpes, we have 2 computers ( a PC & a MAC G4) for data analysis. This keeps the microscopes available for data acquisition.
Coming into confocal as a way to save my position a few years ago, I have only worked with a very old Bio-Rad, which was very reliable, and great for its vintage, and our current Zeiss LSM-510. considering how complex the software must be to do all the things it does, I am always amazed that any of the confocals work. As with any software-driven equipment, you have to expect a few glitches now & again but overall, we have been quite pleased. I can usually get my users up & running fairly independently after about 6-8 hours of training.
Good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
After checking the obvious (filters, water, etc.) try diluting the stain slightly--5-10 % with water. We have had success with this. I suspect the UA (in some cases, at least in ours) may be precipitating out. Also keep the staining dish covered to prevent evaporation (and precipitation). Finally, make sure the drop clinging to the inside of the forceps is drained from behind to prevent it's contaminating the grid.
Sara
On Mon, 2 Jul 2001, Monson, Frederick C. wrote:
} Date: Mon, 2 Jul 2001 12:10:27 -0400 } From: Monson, Frederick C. {fmonson-at-wcupa.edu} } To: 'Eric' {biology-at-ucla.edu} } Cc: 'Microscopy Listserver' {microscopy-at-sparc5.microscopy.com} } Subject: RE: UA ka ka on grids } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Eric, } You said nothing about the solvent.??!!!? While switching out a } filter on the building supply, did someone make an error? Etc. } } You didn't mention the morphology of the ka ka. Smaller than .45um? } } Are these the clumps of homogeneously very small crystals/grains/? } that often appear in older mixes/solutions? } } Has someone else taken sample from the bottle recently? } } Do you do your staining on an open bench? In a hood? In a Petrie } dish? } } Try .22um filter. If still present, then how old is jar of U salt? } } Some other things to think about: } } Biological processes involved with U depletion/removal in } environment may have applicability to lab situations. See: } (http://www.nttc.edu/env/Uranium/Uranium_chap8.html). Almost ALL UAc(Uranyl } acetate) has some trace of Fe in it - increasing the possibility of a slowly } developing linked biological process leading to production of "increasing" } amounts of UO (fine grain ppt?) in the reservoir of UAc. DID you know that } uranium (U) is 'pyrophoric'? } Then there is all that acetate! See: } (http://www.geol.vt.edu/research/gssrs/gssrs2000/abstracts/tkendall.html). } } What's different about the specimen, or is this happening } with ALL specimens? } } [With regard to your concern about your concern. No, you should } never become complacent. Such aggravations must be removed. You must } cultivate your crisis management skills. If you don't, you will feel guilty } and inadequate. It would be better to manifest OCD than to feel 'guilt and } 'inadequacy.'] } } } Regards and best wishes, } } Fred Monson } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI) } Schmucker II Science Center (Room: SS024(Basement)) } South Church Street } West Chester, PA, 19383 } MailDrop: Department of Geology/Astronomy } Phone: 610-738-0437 } Fax: 610-436-3036 } email: fmonson-at-wcupa.edu } Please call before visiting. } } } } } ---------- } } From: Eric } } Sent: Friday, June 29, 2001 6:24 PM } } To: 'Microscopy Listserver' } } Subject: UA ka ka on grids } } Importance: High } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi there fellow microscopists, } } } } Here in the Pathology EM lab we do not do rocket science, but standard } } thin-section TEM of renal biopsies. } } } } In the last two weeks though I have been getting some ka ka (technical } } term) on my grids from my UA stain. I have already made up two new } } batches } } of UA stain and use .45 micron filters just before use. Question is that } } all of the sudden I am getting UA stain artifact on my grids. I have not } } have any trouble since I have been here. } } I have changed everything, new staining plate, cleaned and scrubbed } } everything that I use for staining. I still get some UA artifact. Is } } this } } normal and should I just forget about it or is there something else wrong } } I } } am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I } } stain for usually 10-12 minutes. the grids before staining or clean, but } } after UA stain they are a bit dirty... } } } } Any suggestions? } } } } Thanks, } } } } Eric A. Rosen } } UCLA Medical Center } } Dept. Pathology } } EM Lab } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Good Morning Micha, First, I can only provide my experience on this matter. Second, I will summarize what I have trundled on about below for the purpose of those who are interested in the destination, not the journey.
0. Yes, I have seen "Permount" preps of this age, so I'm certain that mountant, properly applied, will work. The slides I made as a student in 1964 with many different dyes and stains - mountant Gum Damar - are still as 'perfect' as my "B" illustrated. 1. With respect to vapor pressure: Xylene {Toluene {Benzene 2. Three problems with mountants: a. Resin concentration must be close to saturation to minimize evaporative changes in volume after application. b. Mountant volume under cover glass must be close to minimum (I standardize this with weights after application!) c. Resin composition must NOT cause 'unacceptable' changes in specimen (e.g., acidity?).
For years, I have used a natural resin called Gum Damar (or Dammar!) (apparently 'mined' from the same quarter of the planet as hematoxylin), solubilized in Xylene (Xylene {Toluene {Benzene(vapor pressure). This preparation has failed for only one reason that I can discern - too much solvent in the drop(s) used to seal the cover glass. The major problem is loss of solvent via evaporation.
Most of the mountants that are commercially available are solubilized in Toluene, thus, they tend to 'dry' somewhat faster than the prep I have used. While the commercial preparations tend to dry faster, they do not appear to suffer from the other problem apparent in natural preps like Damar that is acidification and/or photochemically induced changes that sometimes lead to bleaching of the dyes or pigments in the mounted specimen.
I have not been in a situation where I needed to preserve a slide past my own estimate of maximum longevity, so Damar has sufficed. I have prepared three liters of Gum Damar since 1967, and the most recent is still fairly full. Each preparation has taken at least a year to finish. I do not expect to make another, and I suspect that you have just decided that you will NOT begin such an ordeal.
So! If you are using Balsam, you will have to wait for 6mos before you can stack the slides as the edge seal takes that long to set.
If you use any mountant with Xylene as the vehicle, you will have the advantage of longevity, but you must use the mountant resin at close to saturation, because that is the equilibrium 'sought' by the mixture and you may have to wait longer for complete setting. Too much vehicle, and the equilibrium volume will be smaller than desired and air may ultimately be drawn into the space between slide and cover glass. The concentration of resin is an important key to long-term success.
The other complication is the volume of the mountant used. This can be solved by careful application of the mountant so that the minimum required volume of saturated mountant is used. Following this, I have always weighted the cover glasses with rectangular lead blocks ( with a square cross section!) measuring 2-3mm less on a side than the cover glass dimensions. Steel blocks cut to appropriate dimensions can be used with fewer parenthetic comments and footnotes and subsequently cleaned when they become contaminated with resin. These weights insure that a standard volume of mountant seals each cover glass. NOTE A REAL DANGER POINT: When training a technician who may perform the bulk of the work, it is important to emphasize the archivists rationale for each step so that when the mountant reservoir (normally in a convenient, but NOT tightly stoppered container becomes too viscous, the proper, minimum, quantity of solvent is added to recalibrate the viscosity. Alternatively, you can choose a better reservoir than that which is commercially available or the dropper bottle that is supplied with some mountants. Do not attempt to seal the open reservoir with grease of any kind. I have often stored the reservoir in a fume hood in a screw capped (cap is Teflon lined!) glass jar, large enough to hold a small narrow-mouth container of vehicle-soaked cotton which I maintain with Pasteur pipetted vehicle. This works nicely when the mountant is not used up rapidly. In situations where a fume hood was NOT available, I have NOT used the jar WITH the cotton-soaked vehicle - just the jar alone! If I am to leave a container of mountant for any length of time, I record the last level and date on a small label applied to the side so that when I take it up again I can easily recalibrate the volume.
There may be some suggestions to paint the edges of the cover glasses. This can, of course, retard the evaporation of the solvent/vehicle, but, since most 'paints' are susceptible to the effects of the organic solvents mentioned here, a retardation is all that can be expected. AND, one must still wait until the edge seal is complete.
When properly prepared, and after edge sealing has completed, a cover glass and slide will behave, in most situations, as a unit object.
For all that you are considering, I can recommend the following book: Lillie, R.D.(1965), Histopathologic Technic and Practical Histochemistry(3d Ed.), McGraw-Hill Book, Co., London [for you], pp91-106 [section on mounting media INCLUDING a brief paragraph on mounting diatoms that is all about indices of refraction]. You will also find much if you search among Gurr's books and papers. When searching the old literature, pause when you find the term, cryptogamic.
My final recommendation is to check in town for a world-famous institute of learning. I bet that if browsing the stacks is still permitted, you can find something of real value in Lee, Vade Mecum(?); Gray, Microtomist's Formulary and Guide; and, again, something by Gurr.
Good luck, archiving is special.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Micha Bayer } Sent: Monday, July 2, 2001 10:52 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: DPX versus Canada Balsam } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } I am in the process of figuring out a way of permanently mounting } desmids (unicellular green algae with a cellulose cell wall) and was } wondering about the relative benefits of DPX and Canada balsam as } mounting media. } } It appears that no-one in our field has done this kind of thing before } (please tell me if I am wrong) and we have to guess our way through } this. } } Is Canada balsam still used? Which mountant has the better long } term and optical characteristics in your experience? Has anyone got } DPX slides which are older than say 20-30 years, and are they still } acceptable? } } many thanks } } Micha Bayer } } ______________________________ } } Dr. Micha Bayer } Royal Botanic Garden Edinburgh } 20A Inverleith Row } Edinburgh EH3 5LR } Scotland, U.K. } Tel. (+44) (0)131-248 2915 or 248 2965 } Fax (+44) (0)131-248 2901 } Project Homepage at http://www.rbge.org.uk/ADIAC/ } RBGE home page at http://www.rbge.org.uk } ______________________________ } }
The book HISTOLOGICAL TECHNIQUES FOR ELECTRON MICROSCOPY by Daniel C. Pease (second edition) published in 1964 by Academic Press, gives a brief history of the use of Toluidine Blue as a stain for survey sections on pages 259-260. It states, "In a symposium discussion of a paper by Mercer (1963), Charles and Meek separately emphasized the usefulness of alkaline toluidine blue. If a 1% solution dissolved in 1% borax is used at 85 degrees C, staining occurs in Araldite in 1-2 minutes."
I hope this helps answer your question.
====================================== Michael D. Standing BYU Microscopy Lab 401 WIDB Brigham Young University Provo, UT 84602
-----Original Message----- } From: Sally Stowe [mailto:STOWE-at-rsbs.anu.edu.au] Sent: Tuesday, July 03, 2001 1:38 AM To: Microscopy-at-sparc5.microscopy.com
Somebody just asked me the reference for toluidine blue/sodium borate (1% each) which is our routine LM stain - I cant find an attribution in any of about 10 books. Can anybody help? This person should not be forgotten! One suggestion was Alsop?
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
} -----Original Message----- } From: Petersen, Maureen A. } Sent: Tuesday, July 03, 2001 11:52 AM } To: 'Microspy-at-MSA.Microscopy.Com' } Subject: TEM available } } List members: } } The Department of Plant Pathology, University of Florida, has a Hitachi } 600 TEM available. } } This microscope has not been on a service contract in recent months, and } went down on me, so I can not say it is currently operating. I THINK I } know where the problem lies, but it needs an engineer. Please contact me } off list, and I will give you more specific information. } } University of Florida labs, then State of Florida labs, have first dibs. } But although the Hitachi has been my companion for years, I recognize that } it may not be a hot item, so no one should hesitate to inquire. } } The microscope is available for the cost of removing it. We have another } scope coming in, so time is limited to arrange to take it. If nothing } else, it could be a good parts scope. } } All interested parties, please contact me ASAP. } } Maureen Petersen } University of Florida } Department of Plant Pathology } Gainesville, FL 32611 } } phone (352) 392-0634
Please tell me more about this stain. Why are you using it and what are it's advantages? Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Sally Stowe [mailto:STOWE-at-rsbs.anu.edu.au] Sent: Tuesday, July 03, 2001 1:38 AM To: Microscopy-at-sparc5.microscopy.com
Somebody just asked me the reference for toluidine blue/sodium borate (1% each) which is our routine LM stain - I cant find an attribution in any of about 10 books. Can anybody help? This person should not be forgotten! One suggestion was Alsop?
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
} I have a Zeiss OPMI surgical microscope. I bought it used as-is. } It came with a bulb that didn't last. The filament broke away } from the support wires. A fine spiral of thinner wire wound around } the support wires seemingly to help support the filament, and it } had spiralled off the supports. } } Fortunately my junk pile revealed a replacement bulb. However, } after the 'scope took a trip in the car to demo for some } kindergarteners, its filament also broke and the thin wires } spiralled away. } } $45 at Bulbdirect.com and I'm back to a working bulb and a spare. } What should I be doing to protect myself against early lamp } failure?
The design of this lamp with the long filament holders makes this lamp prone to filament breakage from impact when the lamp is cold. If you have had problems when transporting the microscope try removing the bulb and storing it in its box. Are you using the 30 watt bulb? It was common for a 50 watt bulb to be installed in this microscope in error.
Cool-down without movement for 20 minutes? NOPE.
Remove } lamps before driving around with the 'scope? YEP.
Are short bursts } of usage (5-10 minutes each) to blame? NOT AT ALL.
Listers, I agree with Richard, it's a parts and service know-how issue. We also have an Amray (1820). Amray didn't discontinue our service, KLA-Tencor did and unfortunately, that's the end of the line for a fine brand of SEM's.
We also have added EDS to our instrument recently and expect to get many more years of service from it. We too, contracted with SEMtech Solutions for service (ex Amray guys) and get service from the same engineer that we had with Amray. A very comfortable situation for us.
If the instrument meets your needs and has parts and service talent available, keep it running as long as it is economical and reliable.
(No vested interest in SEMtech, unless you consider I want them to stay in business so they can fix my instrument!)
Kevin Battjes Impact Analytical Michigan Molecular Institute 1910 W. St Andrews Road Midland MI 48640
-----Original Message----- } From: Bertaska, Richard [mailto:BertaskaR-at-corning.com] Sent: Tuesday, July 03, 2001 7:07 AM To: 'Microscopy-at-MSA.Microscopy.Com'
SEM service life is based on parts availability and support.
Many older SEMs are still supported because the failing parts/components are still manufactured. Fortunately manufacturers used many readily available parts. Also many institutions/businesses have purchased new equipment and "retired the old". Fortunately there are some enterprising individuals who recognized a niche and began collecting "abandoned" SEMs. These individuals now provide a necessary parts/service to SEM industry.
As for support....if you can do much of your own you are smart. For those of us who neither have the time or expertise we need to seek out the experts/entrepreneurs.
I currently manage three SEMs. The oldest is AMRAY1600 purchased in 1984. It is a workhorse for us. Despite minimal maintenance and no service contracts over the past years it has performed well. Recently a number of components began to fail. It was suggested that the system be retired and replaced.
I found expertise in SEMTech Solutions Inc. During the past year we have replaced several aging components, added digital imaging, and moved the SEM to another building for access to multiple users. EDS upgrade is also being considered. As long as parts are available I expect the system to perform another 20 years!
SEMTech provided expertise and parts. I would highly recommend them for AMRAY instrument service.
c/o Jim Peterson PO Box 2155 Natick MA 01760 tel: (978) 663-9822
I have no vested interest in the company. I am simply a truly satisfied customer with an excellent service provider.
Finding other experts for other SEM instrumentation may be difficult. However, if the SEM community needs/supports it, I think there are individuals prepared to provide it.
Perhaps the reason these smaller new EDS ventures have been able to establish themselves is because the larger established businesses no longer listened/responded to their customer needs. A similar circumstance could evolve with SEM maintenance.
The question poses another thought: How could the SEM community "inventory" all SEMs by brand, model, age, location, condition, and status. Thus all SEMs being accounted for could effectively provide the potential inventory parts list and availability.
-----Original Message----- } From: Jensen, Karen Sent: Tuesday, July 03, 2001 2:30 PM To: 'Sally Stowe'
Somebody just asked me the reference for toluidine blue/sodium borate (1% each) which is our routine LM stain - I cant find an attribution in any of about 10 books. Can anybody help? This person should not be forgotten! One suggestion was Alsop?
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
Richardson, K.C., Jarret, L., Finke, E.H.: Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Techn. 35:313, 1960
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Sally Stowe [mailto:STOWE-at-rsbs.anu.edu.au] Sent: Tuesday, July 03, 2001 3:38 AM To: Microscopy-at-sparc5.microscopy.com
Somebody just asked me the reference for toluidine blue/sodium borate (1% each) which is our routine LM stain - I cant find an attribution in any of about 10 books. Can anybody help? This person should not be forgotten! One suggestion was Alsop?
thanks
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
It might be difficult to see the 5 nm gold with SEM (at least in my experience) -- try to put a drop of 1:10 diluted 5 nm gold on a sample holder/stub, air dry and to see if you can identify the gold particles with SEM, if yes, great! if not, try 10 nm or 15 nm. I am sure 15 nm will do it.
Check out the paper:"Localization of myosin on sperm-cell-associated membrances of tobacco" by Zhang Z, HQ Tian and SD Russell on Protoplasma (1999) 208:123-128, in which Immunogold SEM with 15 nm gold is used to localize myosin on cell surface. http://www.springer.at/springer.py?Page=47&Key=87&cat=9&id_abstract=5759&id_ volume=641&id_journal=4
Let me know if you could not find the paper. I will be happy to send you a copy.
Zhaojie Zhang Oklahoma Medical Research Foundation Oklahoma City, OK 73104
-----Original Message----- } From: Mike Delannoy To: microscopy-at-sparc5.microscopy.com Sent: 7/3/01 9:39 AM
Hello Microscopists,
We are doing an IEM for SEM with bacteria and would appreciate any comments. We are looking for surface label 5nm gold with a FIESEM. Our protocol is:
3% PF 0.05% GA fix PO4 buffer
rinse
Bock in 1%NGS
rinse
Primary ABY incubation O/N cold
rinse
Secondary gold conjucated ABY 1hr
rinse
Hard fix 2% GA same buffer
rinse
2% UA filtered DH2O
Dehydration
Critical Point Drying
Our questions are is the UA enough to stablize the lipds throughout the dehydration and cpd? We believe osmium would strip off the gold and tannic acid may make small precipitates similar in size to the gold. We will use in-lens and backscatter to view the gold. Any other suggesstions would be appreciated.
John, My old Zeiss dissecting scope (focal distance about 10" - I love it) has a voltage controller for the lamp. There is a detent in the middle of the range which gives the rated voltage (in this case 6V). It can be considerably over-driven for unusual circumstances, but if you leave it there, very short bulb-life is the result. Check your power supply output.
Ken Converse owner Quality Images third party SEM service Delta, PA
John Foust wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I have a Zeiss OPMI surgical microscope. I bought it used as-is. } It came with a bulb that didn't last. The filament broke away } from the support wires. A fine spiral of thinner wire wound around } the support wires seemingly to help support the filament, and it } had spiralled off the supports. } } Fortunately my junk pile revealed a replacement bulb. However, } after the 'scope took a trip in the car to demo for some } kindergarteners, its filament also broke and the thin wires } spiralled away. } } $45 at Bulbdirect.com and I'm back to a working bulb and a spare. } What should I be doing to protect myself against early lamp } failure? Cool-down without movement for 20 minutes? Remove } lamps before driving around with the 'scope? Are short bursts } of usage (5-10 minutes each) to blame? } } - John } } P.S. Dr. Monson: Love the stories. Keep in mind that all } seemingly newbies on the 'net might be learning very slowly } over time, just for fun, and without benefit of a degree program. } The Web is our Oracle, but I'll try to pay attention during class. } Please do not hit me with eraser, chalk or ruler. } } } }
----- Original Message ----- } From: "Ken Converse" {qualityimages-at-netrax.net} To: "MSA, listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, July 03, 2001 4:15 PM
Hello,
I have a query relating to a project, which is examining the effects of citrus rind oils (mainly terpenoid in composition) on rind tissue. Rind cells appear to 'plasmolyse', membranes degenerate, cell walls collapse and swell. Oil appears to move apo- and symplastically.
My question is: Has anyone examined the effects of similar oils on plant tissues before? Were observations similar to those described above? What is the detailed ultrastructural process of plasmolysis?
Let me add one possible source to the "saw-tooth edge" problem:
60 Hz noise from the power supply (50Hz in other parts of the world).
You can check that by looking up resolution and frame rate and measuring the "wavelength" of the oscillations. Example:
You are looking at 500 lines and 10 frames per second. This gives you around 500 lines per second. A 60 Hz signal should have a "wavelength" of around 8 lines, a 50 Hz signal of around 10 lines ( # of lines x frame rate / frequency). You can also take the measured wavelength and caculate the frequency of the vibration. This can give you some insight into the source of the vibration or interference.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Steve Chapman [mailto:PROTRAIN-at-compuserve.com] Sent: Tuesday, July 03, 2001 7:06 AM To: American Soc
Hi
As the image is in a SEM is created by lines of information, many problems exhibit themselves by giving saw tooth edges to the lines. The slower you scan the better the instrument is at synchronizing out the interference.
The most common reasons for the above are:-
1. Vibration 2. External magnetic fields
If we consider operating parameters we may be able to track this down. Magnetic fields will have a greater affect at lower accelerating voltages and at longer working distances, in both cases the final lens strength is reduced allowing the external field to overcome what protection the lens field provides. Try working at the shortest possible working distance and at the highest kV, probably about 5mm at 30kV. If this reduces or removes the problem it would be an external field that is the culprit. Another indication of an external field problem is the TV rate scan will have a sub ripple that will work its way down the screen.
We need to know if you have been able to take good photographs above 5,000X in the past. If so we then can assume the problem is a result of a change in the laboratory; a new piece of equipment is giving off the field. This may be many feet away, the field effect is according to the inverse square law. Try going round switching off new equipment and see if that helps. I have seen problems due to incorrect wiring of new equipment so if you find a field source have a technician check the wiring before going any further.
Please come back if you need more help.
Steve Chapman Senior Consultant, Protrain For professional training in SEM, TEM and EDX world wide www.emcourses.com
If there are any Philips EM300 series TEM users out there who are in need of sample holders, please contact me. I have four double-tilt holders and a double-tilt LN2 stage that need new homes.
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Another possible source of this is any "spare" oscillators. If The TV scan rate is not generated by the SEM's "main" scan oscillators, those oscillators can sometimes be left running when working in TV frame rate. These oscillators then put noise of their frequency on the power busses, from which it leaks into the scan drive circuits, contaminating the scan. With your scan coils doing "contaminated" scanning, and your monitor or video capture board doing "pure" scanning, you get sawtooth edges. Read the manual very carefully to see if there isn't some way to stop the "main" scan oscillators. Often this info is in the "testing & calibration" section, as you typically need to stop these oscillators to get an "uncontaminated" true reading of the power supply ripple, to verify the supply is OK.
Robert Wieland wieland-at-me.udel.edu No one of us is as smart as all of us. Japanese saying
.. and knowing Murphy, it's probably a combination of more than one effect.
You should be able to test the suggestion below by watching the ripple and change the scan settings on the "spare" oscillator. If the ripple changes, you found the problem.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Robert Wieland [mailto:wieland-at-ME.UDel.Edu] Sent: Wednesday, July 04, 2001 2:09 PM To: Mike Bode Cc: 'Microscopy-at-MSA.Microscopy.Com'
At 07:15 PM 7/4/01 -0700, sammy eze wrote: } ATTN: MANAGING DIRECTOR/CEO. } REQUEST FOR AN URGENT CONFIDENTIAL BUSINESS RELATIONSHIP
Here's the Nigerian Embassy's page on this years-old scam:
http://www.nigerianembassy.nl/letters.htm
This scam accounts for a significant percentage of their GNP.
List Members: Just in case it is not obvious to all, the posting below is a scam. It recently appeared on another list I subscribe to, also.
Maureen Petersen University of Florida
-----Original Message----- } From: sammy eze [mailto:kenno-at-37.com] Sent: Wednesday, July 04, 2001 10:13 PM To: kenayaya-at-amebo.com
Marian: I don't have the references directly available, but the papers of A. Kent Christensen are a very good place to start. I did, however, come across an unattributed method in Hayat's book "Fixation for Electron Microscopy" (badly damaged in a lab flood, but still legible for the most part). Glut (8%) 25ml Paraform (105) 25ml Picric Acid 0.02g 0.1M phosphate buffer to make 100ml (total fixative volume)
Collect first two drops directly into 20ml sol A (above) at RT. Discard proteinaceous seminal plasma which are formed rapidly due to coagulation. Centrifuge the remaining nearly pure cellular suspension at 1000rpm for 10min (total duration of fixation is ~25m). Discard the supernatant and wash the pellet in phosphate buffer for 15m. Postfix the pellet in 1% Os)4 in 0.1M phosphate buffer(pH 7.2) for 15 min at RT.
I have _no_ idea if this is an optimal method, but it's all I can find quickly. Again, I would refer you to Dr. Christensen's work.
Roger On Tue, 03 Jul 2001 13:17:27 -0400, marian miller wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | does someone have a good, and easy method for preserving mouse sperm for | transmission microscopy | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
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Very simple answer, but I, as usual, can't explain it briefly.
We all get married to what we learn originally, because what we learn is the foundation of our individualized empiric relationship with science. Changes have to be couched in terms that are similar to those used to explain our empiric base, OR, we must be forced!
"We use Canada Balsam, because...." Or, as in my case, "I have found Gum Damar to be the best!"
AND, I was just wondering about plastic mountants a minute ago in response to a message from John Bozzola, and was trying, again, to find the specific components of "EPON" and its index of refraction. No success.
Am I familiar with thermoplastics? Yes, from many years ago. The Cargille thermoplastics of today? Not specifically. Reason? Lack of incentive. They were much more expensive, and I was already 'married'. I had 15 year-old preparations with no observable defects, and I was not forced to investigate that alternative for safety reasons either by internal safety flags or external. I had been 'sniffing' for 20 years - one thing and another, and I got 'smarter' every day. ?????
If I had an archival project, such as the one that initiated this thread, I would surely investigate the thermoplastic mountants, in spite of the fact that the cost is MUCH higher/slide. The advantages are obvious.
My reason would be simple, and in concert with your consternation. I would want the insurance of vehicle absence to assure longevity without concern for evaporation and volume shrinkage. I would still minimize usage, but application volumes presumably could be rigidly controlled by applying to each coverglass a pre-determined volume that would form the correct seal, without the otherwise obligatory weight and trimming of excess, and then 'picking' up the cover glass with the inverted slide.
Pathologists would NOT change unless forced to contend with a vehicle/solvent safety issue. Real long-term archiving apparently is NOT the greatest issue with them, and pathology is a REAL customer.
Sometimes, but not often, I am shocked by a loss of memory. In this case, I would certainly abbreviate my previous to Micha even more and suggest that he look carefully at the thermoplastic mountants. Further, now that a safety issue has been raised, I can't think why I would not consider/try them for my routine work.
Thanks for the reminder and the gentle wake-up,
Regards,
Fred Monson
} ---------- } From: Garber, Charles A. } Sent: Tuesday, July 3, 2001 6:22 PM } To: Monson, Fred C. } Subject: RE: DPX versus Canada Balsam } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Thanks for the great lesson, you must be a fantastic teacher in front of a } class. I have a question: Are you familiar with the Cargille Optical } Quality Thermoplastics, as shown on URL } http://www.2spi.com/catalog/ltmic/opt-thermo.html } } Now these are all thermoplastic materials, there is no solvent that can } evaporate. I know that people use these materials for doing some of the } same kinds of mounting you described in your posting. I am trying to } understand why, if the solvent can be the potential cause for so many } problems, why people don't use these optical quality thermoplastics? } } Presumably there is some simple answer, and then I would understand...... } } Chuck } -------- REPLY, Original message follows -------- } } } Date: Tuesday, 03-Jul-01 12:02 PM } } } } From: Monson, Fred C. \ Internet: (fmonson-at-wcupa.edu) } } To: MICROSCOPY BB \ Internet: } } (microscopy-at-sparc5.microscopy.com) } } cc: Micha Bayer \ Internet: (m.bayer-at-rbge.org.uk) } } } } Subject: RE: DPX versus Canada Balsam } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To } } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On- } Line } } Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Good Morning Micha, } } First, I can only provide my experience on this matter. } } Second, I will summarize what I have trundled on about below for } the } } purpose of those who are interested in the destination, not the journey. } } } } 0. Yes, I have seen "Permount" preps of this age, } so } I'm } } certain that mountant, properly applied, will work. The slides I made } as } a } } student in 1964 with many different dyes and stains - mountant Gum Damar } - } are } } still as 'perfect' as my "B" illustrated. } } 1. With respect to vapor pressure: } Xylene {Toluene {Benzene } } 2. Three problems with mountants: } } a. Resin concentration must be close to } saturation } } to minimize evaporative changes in volume after application. } } b. Mountant volume under cover glass must } be } close } } to minimum (I standardize this with weights after application!) } } c. Resin composition must NOT cause } 'unacceptable' } } changes in specimen (e.g., acidity?). } } } } For years, I have used a natural resin called Gum Damar (or } Dammar } !) } } (apparently 'mined' from the same quarter of the planet as hematoxylin), } } solubilized in Xylene (Xylene {Toluene {Benzene(vapor pressure). This } } preparation has failed for only one reason that I can discern - too much } } solvent in the drop(s) used to seal the cover glass. The major problem } is } loss } } of solvent via evaporation. } } } } Most of the mountants that are commercially available are } solubilized } } in Toluene, thus, they tend to 'dry' somewhat faster than the prep I } have } used. } } While the commercial preparations tend to dry faster, they do not } appear } to } } suffer from the other problem apparent in natural preps like Damar that } is } } acidification and/or photochemically induced changes that sometimes lead } to } } bleaching of the dyes or pigments in the mounted specimen. } } } } I have not been in a situation where I needed to preserve a } slide } past } } my own estimate of maximum longevity, so Damar has sufficed. I have } prepared } } three liters of Gum Damar since 1967, and the most recent is still } fairly } full. } } Each preparation has taken at least a year to finish. I do not expect } to } make } } another, and I suspect that you have just decided that you will NOT } begin } such } } an ordeal. } } } } So! If you are using Balsam, you will have to wait for 6mos } before you } } can stack the slides as the edge seal takes that long to set. } } } } If you use any mountant with Xylene as the vehicle, you will } have } the } } advantage of longevity, but you must use the mountant resin at close to } } saturation, because that is the equilibrium 'sought' by the mixture and } you may } } have to wait longer for complete setting. Too much vehicle, and the } } equilibrium volume will be smaller than desired and air may ultimately } be } drawn } } into the space between slide and cover glass. The concentration of } resin } is an } } important key to long-term success. } } } } The other complication is the volume of the mountant used. This } can be } } solved by careful application of the mountant so that the minimum } required } } volume of saturated mountant is used. Following this, I have always } weighted } } the cover glasses with rectangular lead blocks ( with a square cross } section!) } } measuring 2-3mm less on a side than the cover glass dimensions. Steel } blocks } } cut to appropriate dimensions can be used with fewer parenthetic } comments } and } } footnotes and subsequently cleaned when they become contaminated with } resin. } } These weights insure that a standard volume of mountant seals each cover } glass. } } NOTE A REAL DANGER POINT: When training a technician who may perform } the } bulk } } of the work, it is important to emphasize the archivists rationale for } each } } step so that when the mountant reservoir (normally in a convenient, but } NOT } } tightly stoppered container becomes too viscous, the proper, minimum, } quantity } } of solvent is added to recalibrate the viscosity. Alternatively, you } can } } choose a better reservoir than that which is commercially available or } the } } dropper bottle that is supplied with some mountants. Do not attempt to } seal } } the open reservoir with grease of any kind. I have often stored the } reservoir } } in a fume hood in a screw capped (cap is Teflon lined!) glass jar, } large } } enough to hold a small narrow-mouth container of vehicle-soaked cotton } which I } } maintain with Pasteur pipetted vehicle. This works nicely when the } mountant is } } not used up rapidly. In situations where a fume hood was NOT available, } I } have } } NOT used the jar WITH the cotton-soaked vehicle - just the jar alone! If } I } am } } to leave a container of mountant for any length of time, I record the } last } } level and date on a small label applied to the side so that when I take } it } up } } again I can easily recalibrate the volume. } } } } There may be some suggestions to paint the edges of the cover } glasses. } } This can, of course, retard the evaporation of the solvent/vehicle, } but, } since } } most 'paints' are susceptible to the effects of the organic solvents } mentioned } } here, a retardation is all that can be expected. AND, one must still } wait } } until the edge seal is complete. } } } } When properly prepared, and after edge sealing has completed, a } cover } } glass and slide will behave, in most situations, as a unit object. } } } } For all that you are considering, I can recommend the following } book: } } Lillie, R.D.(1965), Histopathologic Technic and Practical Histochemistry } (3d } } Ed.), McGraw-Hill Book, Co., London [for you], pp91-106 [section on } mounting } } media INCLUDING a brief paragraph on mounting diatoms that is all about } indices } } of refraction]. You will also find much if you search among Gurr's } books } and } } papers. When searching the old literature, pause when you find the } term, } } cryptogamic. } } } } My final recommendation is to check in town for a world-famous } } institute of learning. I bet that if browsing the stacks is still } permitted, } } you can find something of real value in Lee, Vade Mecum(?); Gray, } Microtomist's } } Formulary and Guide; and, again, something by Gurr. } } } } Good luck, archiving is special. } } } } Fred Monson } } } } Frederick C. Monson, PhD } } West Chester University of Pennsylvania } } Center for Advanced Scientific Imaging (CASI) } } Schmucker II Science Center (Room: SS024(Basement)) } } South Church Street } } West Chester, PA, 19383 } } MailDrop: Department of Geology/Astronomy } } Phone: 610-738-0437 } } Fax: 610-436-3036 } } email: fmonson-at-wcupa.edu } } Please call before visiting. } } } } } } } ---------- } } } From: Micha Bayer } } } Sent: Monday, July 2, 2001 10:52 AM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: DPX versus Canada Balsam } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } Hi all, } } } } } } I am in the process of figuring out a way of permanently mounting } } } desmids (unicellular green algae with a cellulose cell wall) and was } } } wondering about the relative benefits of DPX and Canada balsam as } } } mounting media. } } } } } } It appears that no-one in our field has done this kind of thing before } } } } (please tell me if I am wrong) and we have to guess our way through } } } this. } } } } } } Is Canada balsam still used? Which mountant has the better long } } } term and optical characteristics in your experience? Has anyone got } } } DPX slides which are older than say 20-30 years, and are they still } } } acceptable? } } } } } } many thanks } } } } } } Micha Bayer } } } } } } ______________________________ } } } } } } Dr. Micha Bayer } } } Royal Botanic Garden Edinburgh } } } 20A Inverleith Row } } } Edinburgh EH3 5LR } } } Scotland, U.K. } } } Tel. (+44) (0)131-248 2915 or 248 2965 } } } Fax (+44) (0)131-248 2901 } } } Project Homepage at http://www.rbge.org.uk/ADIAC/ } } } RBGE home page at http://www.rbge.org.uk } } } ______________________________ } } } } } } } } } } -------- REPLY, End of original message -------- } } }
The MSA/Microsanalysis journal is seeking someone to become part of News and Commentary by writing the "Net Notes" section for the journal. If you are interested in being a part please send reply to JoAn Hudson at hjoan-at-clemson.edu
Thanks to those who pointed out that this is a scam! The two misspellings in the subject heading gave them away (along with pretty much the whole message contained within the email). Maybe they should not have sent it to a list composed of people whose main talent is likely to be well-honed powers of observation and visual data analysis. Kristen
} } From popserve Wed Jul 4 21:23:06 2001 } Date: Wed, 4 Jul 2001 19:03:50 -0700 } X-Mailer: MIME-tools 4.104 (Entity 4.116) } X-Originating-Ip: [63.103.140.102] } From: "sammy eze" {kenno-at-37.com} } To: kenayaya-at-amebo.com } Subject: CONFIDENCIAL BUSSINES PROPOSAL } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
This topic was discussed a few months ago with regards to a question about fixing and embedding human sperm. Here's the result of that, along with the name and information of the generous soul who shared. Kristen
Our lab routinely embeds rabbit and horse spermatozoa. You really will need to centrifuge the ejaculate into a reasonably dense pellet. I wouldn't worry about excess "junk". We fix the ejaculate in 4% glutaraldehyde, 5% sucrose in 1M cacodylate for 24hr. Wash with buffer. Osmicate in 1% OsO4 for 1.5hr. Wash in buffer, centrifuge then embed in 2% low melting point agarose(Sigma A5030), 2% sucrose in buffer. This does very well in 1cc syringes that have been cut off at both ends and the plunger cut even with the bottom so that you can fill them to about 0.5cc and they will fit in a centrifuge tube. You can warm and mix the sperm and agarose in the syringe, keep warm and centrifuge, then push out a perfect pellet. We then use a standard dehydration and epon protocol for TEM. Sorry I don't have published reference for this. I have it as a hand-me down protocol. Jennifer } } ==================================== Jennifer Palmer ARBL Colorado State Univ Ft. Collins, CO 80523-1683 , USA voice:(970)491-1770 fax:(970)491-3557 jpalmer-at-cvmbs.colostate.edu ====================================
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
Thank you Rosemary, I have been looking for that reference for years. I lost it a long while ago, and I think that it IS the ONE!
By the way, I have always thought that 1865 was old! What happened? Also, don't be surprised to discover in that article that the 1% tetraborate buffer wasn't even new then.
For those who want more information, here's a little. [Choice: you can go on with me or divert to a very instructive site: http://members.pgonline.com/~bryand/dyes/class/dyeclass.htm]
There is also a good discussion with many references, though not Rosemary's, in (another OLD book?): Lewis, P.R. and Knight, D.P.(1977), Staining Methods for Sectioned Material, North-Holland Pub. Co., NY,NY, ISBN: 0-7204-0606-4. Part of Series by Audrey M. Glauert, "Practical Methods in Electron Microscopy", same publisher, series ISBN: 0-7204-4250-8.
Toluidine blue O is an thiazine dye that is related to methylene blue(http://members.pgonline.com/~bryand/dyes/azures.htm). Many of these dyes have long been known as "metachromatic" dyes, because they have the property/characteristic ability to provide very nice multi-colored (polychromatic) specimens. The mechanism is NOT perfectly understood, but it is said to rest on the capacity of these dyes to form dye-dye aggregates (pH sensitive because they are charge oriented and the dyes themselves are usually amphoteric) which form generally on more acid and less acid or more basic substances. Everything can be altered by regulating pH. For example, I have used plain 1% aqueous thionin to stain DNA a 'crystal' azure and RNA a muddy purple. I made slides in 1964 that are the same today as when I produced them.
Azure B was once famous for it's use for this purpose. I didn't have any of that, so I used the Organic chemistry that everyone else hated. I think, from my brief, and cursory, comparative studies, that the toluidine blue O/borax mixture 'cooks' much better than most others I tested. Look carefully at the diversity of hue and color in the toluidine blue O (used alone with borate) stained thin sections, and you will see why it was (is with me and others(?)) so popular. I thought everyone knew about it and just chose not to use it, though I have just discovered that Hayat apparently doesn't mention it in his most recent general book on EM, nor in others of his that I have on hand. Don't know why.
Hope this add help.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Rosemary White } Sent: Tuesday, July 3, 2001 4:53 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Tol blue/borax } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Sally, } } In an old (1976) book on botanical microtechnique, there is a reference } for } 1% tol blue in 1% borax - Trump BF, Smuckler EA, Benditt EP (1961) A } method } for staining epoxy sections for light microscopy. J Ultrastr Res } 5:343-348. } } } There's also Mallory's borax methylene blue, maybe tol blue was a } substitute for the methylene blue? } } cheers, } Rosemary } } } Rosemary White } Microscopy Centre } CSIRO Plant Industry } GPO Box 1600 } Canberra, ACT 2601 } Australia } } phone 61-2-6246 5475 or 61-0402 835 973 } fax 61-2-6246 5000 } email r.white-at-pi.csiro.au } } } }
Dear Sally, To address the REAL attribution issue, one of us is going to have to go to a REALLY good library that has these OLD journals back to the late '50's. We may have to divide the labor, but the question is sufficiently interesting and meritorious that we ALL should get it right, when asked. Someone was responsible for the use of borax (Sodium tetraborate) with Toluidine blue O (1% in 1%). Here are all the citations that Pease provides and his [attributions]:
1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc Res., 5: 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in Lewis and Knight below)] 2. Charles and Meek , G.R. separately in a discussion of paper by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O in 1% borax used at 85 degrees C] 3. Pease, D.C.(1964), Histological Techniques for Electron Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 in 2st paragraph the exact method that I use for staining as 'his' method - "...flood the slide, cook until [edge] of drop [froths/]steams [or dries]....". [This is a long discussion to which Pease devotes an entire section, but one can just FEEL his impatience with the unimportance to HIM of this issue. The book, after all, is NOT about light microscopy. The semi-thin section is for orientation only, and hardly deserves any more notice - especially if the light microscopists cannot SEE what is to be seen in the best material ever handed them.] 4. In Kay, D.H.(ed)(1965), Techniques for electron Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this book pre-historic?!] The following is cited as THE reference for using Toluidine blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. cytol, 5: 508.
I have just looked in Geoffrey Meek's book (1970), Practical Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. 456-457. ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he has purposely NOT used citations at all.
I have also consulted the Sorvall manual on "Thin Sectioning"(3d Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated Technics for Electron Microscopy"(1959) which apparently was distributed with the MT-1. In the latter, only staining of Methacrylate sections was given and no mention of 1% *bor* of any kind. In the former, 1% borate at pH 4.5 is mentioned, but that is not relevant to this discussion.
I would bet that you might have some of these references. If you do not, let me know, and I will dig them up, in old Philadelphia, in Ben Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I will do it anyway, because I want to know, but I will do it quicker if you tell me I'm keeping you waiting.
Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Sally Stowe } Sent: Tuesday, July 3, 2001 3:37 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Tol blue/borax } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Somebody just asked me the reference for toluidine blue/sodium borate (1% } each) which is our routine LM stain - I cant find an attribution in any of } about 10 books. Can anybody help? This person should not be forgotten! } One suggestion was Alsop? } } thanks } } Sally } } } } Dr Sally Stowe } Facility Coordinator, } ANU Electron Microscopy Unit } Research School of Biological Sciences } Australian National University } Canberra ACT0200 } AUSTRALIA } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } ph 02 6125 2743 } http://www.anu.edu.au/EMU } } } } }
Question Has anyone seen references or know about research in which dose responses of binding between fluorescently-labeled conjugates and cells have been quantitatively analyzed using light microscopy with a camera detection system and computer software. Thanks in advance Mike D
You might want to consider enhancing the gold with silver, a la, Hayat, M.A.(1995), Immunogold-Silver Staining, etc., CRC Press, Boca Raton, FL, ISBN: 0-8493-2449-1. LC #: QR187.I482H39. I haven't done that, however.
Comforted that you are going to use BSD, I have used it for Ag impregnated intercellular spaces, but the Ag concentrations were quite high. Can you do EDS mapping as well?
Other source: http://www.ebsciences.com/papers/immusem.htm
Hermann R, Schwarz H, Müller M (1991). High precision immuno-scanning electron microscopy using Fab fragments coupled to ultra-small colloidal gold. J. Struct. Biol., 107(1): 38 - 47. Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Mike Delannoy } Sent: Tuesday, July 3, 2001 10:39 AM } To: microscopy-at-sparc5.microscopy.com } Subject: IEM for SEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Microscopists, } } We are doing an IEM for SEM with bacteria and would appreciate any } comments. We are looking for surface label 5nm gold with a FIESEM. Our } protocol is: } } 3% PF 0.05% GA fix PO4 buffer } } rinse } } Bock in 1%NGS } } rinse } } Primary ABY incubation O/N cold } } rinse } } Secondary gold conjucated ABY 1hr } } rinse } } Hard fix 2% GA same buffer } } rinse } } 2% UA filtered DH2O } } Dehydration } } Critical Point Drying } } Our questions are is the UA enough to stablize the lipds throughout the } dehydration and cpd? We believe osmium would strip off the gold and } tannic acid may make small precipitates similar in size to the gold. } We will use in-lens and backscatter to view the gold. Any other } suggesstions would be appreciated. } } Thank you } } Mike D } } }
Does anyone have experience with embedding, in resin, tissue fixed in 4%PF that was labelled with an alkaline phospatase secondary ab reacted with the substrate BCIF/NBT. The concern is the fading/dissolving/diffusion of the substrate during processing.
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
I wrote about modifying a Nikon Diaphot 80/20 prism and received some good suggestions (the 100/0) prism is no longer available from Nikon). When I took the cover off the side of the microscope and removed the prism, I discovered that one solution might be simply covering the binocular output surface of the 80/20 prism. Would there be a problem with this approach? I realize that I would only have 80% going to the specimen but that may be tolerable. What would be the best way to block the laser beam from exiting from the 20% prism surface? Black tape? Black paint? Black metal ? Silver tape covered with black tape?
The following position is open at Honeywell Engines and Systems in Phoenix, AZ.
Microscopy Engineer/Technician
Honeywell Engines and Systems Materials Laboratory is looking for an experienced engineer/technician knowledgeable in the operation of an electron microprobe (EPMA) and scanning electron microscope (SEM) to support production and development of aerospace materials. In depth knowledge of and micro-x ray analysis methods using EDS and WDS is required.
Contacts:
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or
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Your question is a good one and I also would be interested to know of any experience with this. I know that Vector Red (another alkaline phosphatase substrate) can be dehydrated through ethanol and the tissue cleared and it is supposed to survive. One would think then that embedding should also be possible. If epoxy gives a problem perhaps LR white??
We have a Dynavac Xenosput Cr coater which has recently used up the last of the Ti in the Ti sub pump target. When we ordered 2 replacement targets (with part numbers specified) from a local dealer (I don't want to mention company names) we received two pieces of metal (presumably Ti) which had been cut from a sheet of metal using an oxy/acetylene torch. The pieces were not finished in any way. Needless to say we were not overly impressed!!
Does anyone know of sources (either local or international) for these targets?
Many thanks for any information.
Colin Veitch
Instrumentation Scientist Late Stage Innovation Group CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
To contribute to the search for the roots of the Toluidine blue procedure:
There is an interesting paper by Kramer & Windrum (J. Histochem Cytochem, 1955, Vol 3, 227-237, The Metachromic Staining Reaction) that probably was instrumental in initiating the wider use of Tol Blue. In this paper the effect of stain concentration and residual water is shown (amongst many other factors) to be important for the development of the multiplicity of colours.
The use of Toluidine Blue as a stain for monitor sections of epoxy- and acrylic-embedded tissue probably stems from these metachromic effects that this stain produces, with the result that some deductions regarding the chemical composition of the sample can be made after a simple Tol Blue staining.
Even though we, in this lab, have since forever used Tol Blue for the staining of monitor sections of resin-embedded tissues, every batch of Tol Blue that we make up seems to have slightly different staining characteristics. This was such a source of irritation that Chris van der Merwe recently decided to systematically investigate some of the variables.
He found that the most important parameters are: 1. Tol blue concentration (0.1 or 0.5%) 2. Buffer or vehicle type, pH and concentration (0.1 - 1% Na-carbonate, Na-benzoate buffer at pH4, Na-tetraborate (borax) at pH 8.8) 3. Embedding resin formulation (Spurr, Quetol, Polarbed 812, LR-White) 4. Section thickness.
All the above have an effect on the intensity, as well as the degree of metachromicity, of the stain. Resin formulation is probably the major influencer of the results.
This may be a reason for the existence of the wide variety of Tol Blue recipes that abound in the literature.
More information on Tol Blue staining can be had directly from Chris at cvdmerwe-at-scienti.up.ac.za
Regards
Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
"Monson, Frederick C." wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Sally, } To address the REAL attribution issue, one of us is going to have to } go to a REALLY good library that has these OLD journals back to the late } '50's. We may have to divide the labor, but the question is sufficiently } interesting and meritorious that we ALL should get it right, when asked. } Someone was responsible for the use of borax (Sodium tetraborate) with } Toluidine blue O (1% in 1%). Here are all the citations that Pease provides } and his [attributions]: } } 1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc Res., 5: } 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in Lewis } and Knight below)] } 2. Charles and Meek , G.R. separately in a discussion of paper } by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O in } 1% borax used at 85 degrees C] } 3. Pease, D.C.(1964), Histological Techniques for Electron } Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 in } 2st paragraph the exact method that I use for staining as 'his' method - } "...flood the slide, cook until [edge] of drop [froths/]steams [or } dries]....". [This is a long discussion to which Pease devotes an entire } section, but one can just FEEL his impatience with the unimportance to HIM } of this issue. The book, after all, is NOT about light microscopy. The } semi-thin section is for orientation only, and hardly deserves any more } notice - especially if the light microscopists cannot SEE what is to be seen } in the best material ever handed them.] } 4. In Kay, D.H.(ed)(1965), Techniques for electron } Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this book } pre-historic?!] The following is cited as THE reference for using Toluidine } blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. } cytol, 5: 508. } } I have just looked in Geoffrey Meek's book (1970), Practical } Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. 456-457. } ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he has } purposely NOT used citations at all. } } I have also consulted the Sorvall manual on "Thin Sectioning"(3d } Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated } Technics for Electron Microscopy"(1959) which apparently was distributed } with the MT-1. In the latter, only staining of Methacrylate sections was } given and no mention of 1% *bor* of any kind. In the former, 1% borate at } pH 4.5 is mentioned, but that is not relevant to this discussion. } } I would bet that you might have some of these references. If you do } not, let me know, and I will dig them up, in old Philadelphia, in Ben } Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I will } do it anyway, because I want to know, but I will do it quicker if you tell } me I'm keeping you waiting. } } Regards, } } Fred Monson } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI) } Schmucker II Science Center (Room: SS024(Basement)) } South Church Street } West Chester, PA, 19383 } MailDrop: Department of Geology/Astronomy } Phone: 610-738-0437 } Fax: 610-436-3036 } email: fmonson-at-wcupa.edu } Please call before visiting. } } } } ---------- } } From: Sally Stowe } } Sent: Tuesday, July 3, 2001 3:37 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Tol blue/borax } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Somebody just asked me the reference for toluidine blue/sodium borate (1% } } each) which is our routine LM stain - I cant find an attribution in any of } } about 10 books. Can anybody help? This person should not be forgotten! } } One suggestion was Alsop? } } } } thanks } } } } Sally } } } } } } } } Dr Sally Stowe } } Facility Coordinator, } } ANU Electron Microscopy Unit } } Research School of Biological Sciences } } Australian National University } } Canberra ACT0200 } } AUSTRALIA } } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } } ph 02 6125 2743 } } http://www.anu.edu.au/EMU } } } } } } } } } }
Continuing on this thread, I had always thought Richardson's stain was the methylene blue/azure II combination, but not so, it seems, since the 1960 paper refers to both this stain and to tol blue. Someone emailed that methylene blue must be used with azure II, but I don't think this is necessary, though it may give better staining.
Re metachromicity, as Fred Monson and Jan Coetzee note, it depends quite a bit on pH of the tol blue (for plant material at least), and used to depend on the source of stain but I guess this is not such a problem these days(?). We generally use a benzoate buffered solution of tol blue at pH 4.5 for fresh tissue and tissue embedded in hydrophilic resins like LR White and other methacrylates, and a pH 10 solution (buffered, if necessary, in sodium carbonate) for sections of hydrophobic epoxy resins - e.g. Spurr's. Tol blue metachromicity isn't so good at high pH (at least in my hands). The pH of tap water is usually slightly acidic, so aqueous tol blue is generally OK for fresh tissue and methacrylate resins. Since borate buffers are alkaline, perhaps this was the original reason they were used for the hydrophobic resins(?) - and they do produce a much more permanent stain.
Re. "old" references and books, most people here do molecular/genetic type work and literature older than about 1995 is considered venerable..... I am amazed (and appalled) at the great old histochemistry and cytochemistry books thrown out by libraries - the source of most of my "old" books.
Friday night, burning a few CDs..... Looked up another reference on histochemistry - A.G. Everson Pearse (1968) Histochemistry Vol 1 3rd edn (SBN 7000-1326-1 for Fred Monson - perhaps ISBN came after this time....) - a method from 1947 is given that uses a solution of 0.25% tol blue in 0.25% borax followed by 0.5% potassium dichromate in sat'd mercuric chloride, for permanent staining of paraffin sections prior to dehydration and mounting in Canada Balsam. Hess M, Hollander F (1947) J Lab Clin Med 32:905
Mallory, whose name used to be associated with various staining protocols, put it all together in a book: Mallory FB (1938) Pathological Technique. WB Saunders, Philadelphia. This is where Mallory's borax methylene blue comes from - 1% methylene blue plus 1% azure II in 1% borax. Lillie refers to it in his book - Histopathologic Technic - we have the 2nd edition, published in 1954, LC number 53-5622 - again, pre-ISBN.
Will finish now, in case this is getting too bor-ing!
cheers, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
I have a Link detector with AN1000 electronics and a Dapple Systems analyzer. This has been a very stabile system for many years. I recently lost count rate with a broad peak near zero. I ran a few cycles with my conditioner and then was able to see ice at the bottom of the dewar (it has been humid here). I proceeded to empty the dewar, dry it out with alcohol and acetone and re-install.
I still have a low count rate and signal noise. Can anyone offer some pointers or know of a local service engineer. I am in the Chicago area.
Dear Hank, There are lots of books that explain the histochemical methods for alkaline phosphatase, but few that concentrate on EM. NBT is converted to a formazan that is insoluble, but not electron dense. I'm not certain about the effect of OsO4 on the formazan but it has been used to osmicate the ppt when DAB is used. There is no need to go further here, except that phosphatase histochemistry at the EM level has been accomplished by using metal capture methods for some time.
I recommend a book that has the information you need beyond what I have given above. One with protocols is:
Ogawa, K. and Barka, T.(1993), Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine, CRC Press, Boca Raton, FL, ( for the ILL reprint: pp. 140-144("Alkaline Phosphatase")), ISBN: 0-8493-6012-9, LC Card #: 92-9241, LC RB46.7.E43 (In the library). In truth, this book should be in the library at Baylor. If not, perhaps you might get it ordered. It is not out of date. Most of the recent (since '80's) innovations in enzyme histochemistry have concerned buffering and fixing to retain maximum enzymatic activity while not sacrificing structural integrity. In addition, as in LM, you will need to use controls for the EM specimens.
You might also want to consider something like the following protocol/approach.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: hpadams } Sent: Thursday, July 5, 2001 4:10 PM } To: microscopy-at-sparc5.microscopy.com } Subject: bcip/NBT substrate and resin } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have experience with embedding, in resin, tissue fixed in 4%PF } } that was labelled with an alkaline phospatase secondary ab reacted with } the } substrate BCIF/NBT. The concern is the fading/dissolving/diffusion of the } substrate during processing. } } Hank Adams } Manager } Integrated Microscopy Core } Molecular and Cellular Biology } Baylor College of Medicine } Houston, Tx 77030 } } }
Dear Damian, Seems to me you have gotten to the nexus of the problem. Which optical path do you want? Straight ahead or 45 degrees. If straight ahead, store the part and cover the opening. If 45 degrees then spend a few dollars (i.e., http://www.mwkindustries.com/A-f/miru1.htm + a 45degree block and rod) so you can get all 100%. Then, when the current project is over, you can put the scope back together so someone else can use the 80/20. I'm assuming, from what you say that you don't want to use the oculars at all - ever.
Regards,
Fred Monson
} ---------- } From: Damian } Sent: Thursday, July 5, 2001 5:24 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Modifying a Prism } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I wrote about modifying a Nikon Diaphot 80/20 prism and received some good } } suggestions (the 100/0) prism is no longer available from Nikon). When I } took the cover off the side of the microscope and removed the prism, I } discovered that one solution might be simply covering the binocular output } } surface of the 80/20 prism. Would there be a problem with this } approach? I realize that I would only have 80% going to the specimen but } that may be tolerable. What would be the best way to block the laser beam } } from exiting from the 20% prism surface? Black tape? Black paint? Black } } metal ? Silver tape covered with black tape? } } Thanks for the help } } Damian } } }
ProSciTech was not THAT supplier. We have excellent suppliers for targets, including Ti targets - I have no idea about the Xenosput target dimensions. Targets seem to be a simple product but there are great price and quality variations, yet high price frequently does not result in high quality. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, July 06, 2001 2:37 PM, "colin.veitch-at-tft.csiro.au"-at-sparc5.microscopy.com [SMTP:"colin.veitch-at-tft.csiro.au"-at-sparc5.microscopy.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } We have a Dynavac Xenosput Cr coater which has recently used up the last of } the Ti in the Ti sub pump target. When we ordered 2 replacement targets } (with part numbers specified) from a local dealer (I don't want to mention } company names) we received two pieces of metal (presumably Ti) which had } been cut from a sheet of metal using an oxy/acetylene torch. The pieces } were not finished in any way. Needless to say we were not overly } impressed!! } } Does anyone know of sources (either local or international) for these } targets? } } Many thanks for any information. } } Colin Veitch } } Instrumentation Scientist } Late Stage Innovation Group } CSIRO Textile and Fibre Technology } PO Box 21, BELMONT, Vic. 3216. Australia. } } E-mail: colin.veitch-at-tft.csiro.au } Web: http://www.tft.csiro.au } } Tel: +61 (0) 3 5246 4000 } Fax: +61 (0) 3 5246 4811 } } } The information contained in this e-mail message may be privileged or } confidential information. If you are not an intended recipient, you may not } copy, distribute or take any action in reliance on it. If you have received } this message in error, please telephone CSIRO Textile and Fibre Technology } on +61 3 5246 4000.
If they are in a format such as TIFF that can be opened with a picture editing programs like Photoshop (including Photoshop LE) or NIH image or Paintshop Pro or probably others, just change the color mode to indexed color and apply appropriate color palette or LUT - they come with samples palettes. Paintshop Pro has the advantage of using ASCII format palettes which can be made in Excel (Red, Green Blue values 0-255) and imported.
} From: "Bharesh_Mandalia-at-gillette.com"-at-sparc5.microscopy.com } Subject: False Colour SEM Images } To: Microscopy-at-sparc5.microscopy.com } X-Mailer: Lotus Notes Release 5.0.6 December 14, 2000 } Date: Fri, 6 Jul 2001 07:36:57 +0100 } X-MIMETrack: Serialize by Router on GBO827N/BO/Gillette(Release 5.0.5 } |September 22, 2000) at } 07/06/2001 02:34:29 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
----------------------------------------------------------- Glenn Holm {mailto:gholm-at-mit.edu} Graybiel Lab (617)253-5780;fax (617)253-1599 M.I.T Dept. of Brain + Cog. Sci. Cambridge, MA 02139 ----------------------------------------------------------
We're looking for suggestions for SEM standards to calibrate brightness/contrast settings (for example, setting an auto B/C feature in the SEM software, so that the desired repeatable B/C level is obtained by multiple users). Our customers' goal is to do correction/matching of gray scale histograms between images of different samples, and they would like to make the process of adjusting B/C levels at the start of image acquisition to be as standardized as possible. The imaging might be done in secondary or backscattered mode, so suggestions for both would be appreciated.
Thanks vey much for you input,
Elaine
Elaine F. Schumacher Analytical Specialist UOP LLC 25 East Algonquin Road Des Plaines, IL 60017-5017 Phone: 847-391-3403 Fax: 847-391-3719 Email: efschuma-at-uop.com
Adobe photoshop is an excellent application for manipulating both SEM and TEM images. Conversion of gray scale images to color requires a little practice with the software but the results are well worth the effort. I have also used Adobes Illustrator to design and produce presentation posters of my images.
Pete Polsgrove pjp6-at-dana.ucc.nau.edu
} ===== Original Message From "Bharesh_Mandalia-at-gillette.com"-at-sparc5.microscopy.com ===== ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Is there a way to convert greyscale SEM images to colour, like the ones at the MicroAngelo website?
} Is there a way to convert greyscale SEM images to colour, like the ones at } the MicroAngelo website?
I personally "paint" each SEM image with Photoshop, rather than use lookup tables. For TEMs, though, I sometimes use LUTs.
I have a graphics tablet and stylus and I enlarge each image so that I can practically see each pixel and I apply color with the paintbrush in Color mode, so this is a slightly more "artistic" endeavor, rather like paint-by-numbers! It takes hours and hours for each image. However, I have seen some great results with applying LUTs, which is certainly a lot faster!
Experiment. Have fun!
Aloha, Tina (MicroAngela)
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Dear Alan, It is probably worth getting your detector serviced. My experience with ice in the dewar was that I had to warm it up thoroughly to get rid of ice that had condensed inside the crystal/preamp part, where you can't get at it with solvents and then the moisture in the vacuum caused poor holding time for the LN2, so I had the dewar pumped. Jim Nicolino of AAT (JNicolino-at-xraydetectors.com), which services all makes of detectors, says it is worth getting the whole thing properly, professionally checked out, the molecular sieve replaced and the detector brought back up to factory specs. He is probably right. I have no affiliation with AAT except as a satisfied customer. At 05:56 AM 7/6/01 -0500, you wrote: } } I have a Link detector with AN1000 electronics and a Dapple Systems } analyzer. This has been a very stabile system for many years. I recently } lost count rate with a broad peak near zero. I ran a few cycles with my } conditioner and then was able to see ice at the bottom of the dewar (it has } been humid here). I proceeded to empty the dewar, dry it out with alcohol } and acetone and re-install. } } I still have a low count rate and signal noise. Can anyone offer some } pointers or know of a local service engineer. I am in the Chicago area. } } Alan Stone } ASTON Metallurgical Services } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hi All: I was told that there exists image matching software that allows one to calibrate the image one wants for the specific paper one is printing on.
Has anyone used this type of software and if so, have any recommendations The less expensive, the better? Thanks, Mike Coviello UTA
Unfortunately it may not be as easy as that. If you change the LUT, the changes are applied to all pixels of the image. If your object has similar gray values than other parts of the image, you end up with colors all over the image.
You basically have to segment the image first into parts that you want to change and parts you want to stay untouched, then color one part (for example using the LUT functions), then put the two parts together again.
If you want to get some recipes on how to do this, you might want to ask Dr. Gary Gaugler about it. He has used our software (analySIS) and other to the kind of coloring you are interested in. Go to www.photoweb.net to find out more.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Glenn Holm [mailto:gholm-at-mit.edu] Sent: Friday, July 06, 2001 9:20 AM To: Microscopy-at-sparc5.microscopy.com
If they are in a format such as TIFF that can be opened with a picture editing programs like Photoshop (including Photoshop LE) or NIH image or Paintshop Pro or probably others, just change the color mode to indexed color and apply appropriate color palette or LUT - they come with samples palettes. Paintshop Pro has the advantage of using ASCII format palettes which can be made in Excel (Red, Green Blue values 0-255) and imported.
} From: "Bharesh_Mandalia-at-gillette.com"-at-sparc5.microscopy.com } Subject: False Colour SEM Images } To: Microscopy-at-sparc5.microscopy.com } X-Mailer: Lotus Notes Release 5.0.6 December 14, 2000 } Date: Fri, 6 Jul 2001 07:36:57 +0100 } X-MIMETrack: Serialize by Router on GBO827N/BO/Gillette(Release 5.0.5 } |September 22, 2000) at } 07/06/2001 02:34:29 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
----------------------------------------------------------- Glenn Holm {mailto:gholm-at-mit.edu} Graybiel Lab (617)253-5780;fax (617)253-1599 M.I.T Dept. of Brain + Cog. Sci. Cambridge, MA 02139 ----------------------------------------------------------
Hi Liststers! I would appreciate any information on the safety of hexamethyldisilazane (HMDS). I didn't get much out of the data sheet. I would like to know what I am working with... Is it hazardous to one's health? I don't have a hood in my room, but I could move to another room which does have a hood. Thanks!
We always use HMDS in a hood. I'm home and info is at work though.
Ron ----- Original Message ----- } From: "Barbara Plowman" {Bplowman-at-sfmail.dental.uop.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, July 06, 2001 7:18 PM
Dear ALL,
Is there a version of MAcTempas/CrystalKit for a Windows/PC platform?
Is there nay other windows platform Package doing a similat job?
The nearest program in the PC world is JEMS. I recently examined what was available from PC's particularly a program that could simulate electron diffraction.
I purchased Pierre Stadelman's JEMS - runs in JAVA - and it is beautiful for both images and diffraction. Runs very nicely on a Compaq 1.6 GHz machine with 512 MB and will also run on a Mac or Sun. I have already set up all the amphibole minerals mainly for electron diffraction work. Very useful so far. JEMS also plays well with triclinic phases something many other programs have difficulty with. Something to do with right angles I guess.
Check it out at {http://cimewww.epfl.ch/people/stadelmann}
MacTempas and CrystalKit together were(?) unique however in how they were able to treat interfaces such as twins. I don't think JEMS does this very easily.
I found however that if you are interested in looking at crystal structures neither CrystalKit nor JEMS can compete with David Palmer's CrystalMaker program which currently runs only on a Mac. He is however working on a C++ version that will run on PC's.
Check it out at {http://www.crystalmaker.co.uk} and email him that you are anxiously awaiting the PC version.
My advice is purchase an $800 iMac and run the Mac programs and give JEMS a good look for your PC. Can't have too many platforms.
Cheers, Gordon Nord US Geological Survey - Emeritus
Maria Lada wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear ALL, } } Is there a version of MAcTempas/CrystalKit for a Windows/PC platform? } } Is there nay other windows platform Package doing a similat job? } } with thanks } } Maria Lada } EEE department } University of Sheffield
HMDS *MUST* be used in a hood. I'm at home as well, so don't have the MSDS, but there is no question about this.
Phil
} Hi Liststers! } I would appreciate any information on the safety of } hexamethyldisilazane (HMDS). I didn't get much out of the data } sheet. I would like to know what I am working with... Is it } hazardous to one's health? I don't have a hood in my room, but I } could move to another room which does have a hood. Thanks!
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Maria, Thierry Epicier has written SIMPLY-S code for HREM image simulation (and more) for the PC. The simulation part is based on the original SHRLI (simulated high-resolution lattice image) code. Since MacTempas is a port to the Mac of the TEMPaS code (developed from SHRLI to run on a Vax system with a Tektronics GUI), the results of SIMPLY-S and MacTempas should be the same. You can reach Thierry at Thierry.Epicier-at-insa-lyon.fr Mike O'Keefe
----- Original Message ----- } From: "Maria Lada" {m.lada-at-sheffield.ac.uk}
Hi Barbara,
I received my HMDS from Agar Scientific and it came with a 6 page material safety data sheet.
In short:
it is highly flammable, toxic, harmful by inhalation, in contact with skin and if swallowed, irritating to eyes, causes severe skin irritation, is readily absorbed through the skin - and the target organ is nerves.
In case of contact with eye, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing.
You find MSDS in the internet by doing a keyword search and the major EM suppliers have MSDS of their products on their web sites.
I always use it in the fume cupboard, with gloves - and make sure that their is no hot plate near by. (in fact I handle it as I would handle Propylen oxide).
Regards
Claudia
Date sent: Fri, 06 Jul 2001 16:18:12 -0700 } From: Barbara Plowman {Bplowman-at-sfmail.dental.uop.edu} To: Microscopy {Microscopy-at-sparc5.microscopy.com}
Hi Liststers! I would appreciate any information on the safety of hexamethyldisilazane (HMDS). I didn't get much out of the data sheet. I would like to know what I am working with... Is it hazardous to one's health? I don't have a hood in my room, but I could move to another room which does have a hood. Thanks!
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk Fax: 44(0)208 547 7562
Baresh, there is a very easy, quick and effective way if you have Adobe PhotoShop:
1) Create a new layer above your image. 2) Select the region you want to colour. 3) Fill it with the colour you want. 4) Change the layer properties from 'Normal' to Overlay. Done!
You may do step (2) by drawing round the region with the 'freehand select' or 'polygon select' tool, or on the lower layer using the magic wand.
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone." Marconi Optical Components Limited Registered in England No. 4113798 Registered Office: One Bruton Street London W1J 6AQ
A BSE standard can be made from almost any materials (suitable for SEM). The std should be ground and polished to remove topography contrast. What materials you choose will depend on the examination goals.
This brings me to a point. I would not stress BSE "calibration" as a generally useful technique. Under VERY limited circumstances, such calibration/standardization can be beneficial. Routine use of this technique will guarantee that sooner or later an important phase will be missed when examining an unknown. When examining with BSE, I find it imperative to be "all over the map" with the BSE B&C to be sure I am not missing something. It is not unheard of to need two or more images to define what is happening. Sometimes there is simply not enough dynamic range presented in one image to perceive all variations in composition.
Regards, Woody White, McDermott Technology, Inc.
} -----Original Message----- } From: Schumacher, Elaine [mailto:efschuma-at-uop.com] } Sent: Friday, July 06, 2001 12:27 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Brightness/Contrast Calibration Standards } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Fellow Microscopists, } } We're looking for suggestions for SEM standards to calibrate } brightness/contrast settings (for example, setting an auto } B/C feature in } the SEM software, so that the desired repeatable B/C level is } obtained by } multiple users). Our customers' goal is to do } correction/matching of gray } scale histograms between images of different samples, and } they would like to } make the process of adjusting B/C levels at the start of } image acquisition } to be as standardized as possible. The imaging might be done } in secondary } or backscattered mode, so suggestions for both would be appreciated. } } Thanks vey much for you input, } } Elaine } } Elaine F. Schumacher } Analytical Specialist } UOP LLC } 25 East Algonquin Road } Des Plaines, IL 60017-5017 } Phone: 847-391-3403 } Fax: 847-391-3719 } Email: efschuma-at-uop.com } }
Dear Smartech, If someone published such an article comparing EDS detectors as to their ability to detect light elements it should be the SEM manufacturers. The light element sensitivity is determined by the geometry of the detector not the entrance window. Basically the higher the take-off angle of the emerging photon, the better the sensitivity due to the absorption of the matrix. The high voltage of the electron beam also changes the sensitivity due to the depth of penetration and therefore the depth of the x-ray source. Operating a EDX repair facility, I work on all of the EDX manufacturer detectors. With one possible exception, all the manufacturers use the same entrance window material, using various trade mark names. There also seems to be very little difference is the Si(Li) crystals or FET's which are the two internal components of the detectors. Since all of the manufacturers use a resolution specification for Fe55 photons and a peak to background ratio for the 1.0 Kev channel, there would be no comparison specifications for light element sensitivity. I hope this answers your question. Jim Nicolino
I concur with Woody's comments, and offer a couple of more.
It is often possible for the same sample type to set BSE brightness to be consistent between images, and there are many time you would want to do that. We do image analysis of concrete and use calcite and graphite to repeatably set our signal. Some images look good that way, but even then, variation in polishing can greatly affect what the final image looks like. Sometimes it is not so good.
Last week I was looking at samples of a Cr-Al alloy. (It required much different contrast than did the concrete.) Some areas of some samples had three phases present (Al, Al-Cr, and Cr); others had only two (Al and Al-Cr). My inclination was naturally to first turn up the contrast to spread the signal throughout the grayscale to show the character of what was present. I later went back to take some images at consistent signal levels to show that the Cr-rich phase was missing.
Now "calibration" is a reasonable process for BSE. It is much less tractable for SE. SE signals are just not that consistent. Since they are so sensitive to topography, I find myself changing contrast all the time. I expect to need to reduce contrast as I increase magnification as more details come into view. That is the nature of it.
I wonder, do you have a waveform monitor on your scope to help in setting contrast? We use ours religiously and I have been surprised to hear that some scopes don't have them. (I guess the one we had on our JEOL U3 was an add-on, but since they have been part of our spec.) We have determined where white and black are on our waveform display, and before each photo we check the waveform to be sure that the signal is within our recordable range.
There is still some experience involved in setting the level so the picture looks good afterwards. A picture may be completely within the recordable range and still "look" too dark or light. I tend to prefer my background away from full black or white even if there is nothing to see in it. That just takes practice since there is such a wide variety of samples out there.
Digital imaging with gamma control after the fact has done a lot to make sure the first picture will be a keeper. If you don't have that yet, please consider it, or figure that your operators will just have to practice more.
Cheers, Warren
At 07:18 AM 7/9/2001 -0500, you wrote: } Elaine, } } A BSE standard can be made from almost any materials (suitable for SEM). } The std should be ground and polished to remove topography contrast. What } materials you choose will depend on the examination goals. } } This brings me to a point. I would not stress BSE "calibration" as a } generally useful technique. Under VERY limited circumstances, such } calibration/standardization can be beneficial. Routine use of this } technique will guarantee that sooner or later an important phase will be } missed when examining an unknown. When examining with BSE, I find it } imperative to be "all over the map" with the BSE B&C to be sure I am not } missing something. It is not unheard of to need two or more images to } define what is happening. Sometimes there is simply not enough dynamic } range presented in one image to perceive all variations in composition. } } Regards, } Woody White, McDermott Technology, Inc. } } } -----Original Message----- } } From: Schumacher, Elaine [mailto:efschuma-at-uop.com] } } Sent: Friday, July 06, 2001 12:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Brightness/Contrast Calibration Standards } } } } Fellow Microscopists, } } } } We're looking for suggestions for SEM standards to calibrate } } brightness/contrast settings (for example, setting an auto } } B/C feature in } } the SEM software, so that the desired repeatable B/C level is } } obtained by } } multiple users). Our customers' goal is to do } } correction/matching of gray } } scale histograms between images of different samples, and } } they would like to } } make the process of adjusting B/C levels at the start of } } image acquisition } } to be as standardized as possible. The imaging might be done } } in secondary } } or backscattered mode, so suggestions for both would be appreciated. } } } } Thanks vey much for you input, } } } } Elaine } } } } Elaine F. Schumacher } } Analytical Specialist } } UOP LLC ---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The main tricks are to low-angle mill both sides of the specimen and to not use Cu grids. We switched to Mo grids years ago and we rarely see back sputtered material on samples milled on both sides (from the substrate direction, of course). If you want to use up your stock of Cu grids, cut the back side of the grid away so that there is no grid material beyond the specimen to sputter from. Grid becomes the "C" shaped grid used for FIB prepped specimens. Sometimes some sputtered material persists after very low angle milling. In that case, pop the sample into a conventional high-angle mill and mill at 25 degrees for 30 seconds at 2 or 3 KeV to clean off the sputter artifact.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Richard Beanland +44 1327 356363 {richard.beanland-at-marconi.com} on 07/02/2001 07:02:41 AM
To: Feng Wu {fwu-at-bgumail.bgu.ac.il} cc: Microscopy Listserver {microscopy-at-sparc5.microscopy.com}
Feng, the trick is not to have a hole in your specimen where the ions can go through and back-sputter the Cu on to your sample. So make your specimen big enough to cover up the hole in the support grid and stop ion milling immediately when a hole appears. The problem will go away to some extent if you use the two-sided specimen holder. Alternatively, use an inert grid and dissolve the Cu with an acid after specimen prep. (Most acids are unlikely to affect GaN or sapphire.)
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } }
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Another potential cause of variation is dye concentration. To be certified by the Biological Stain Commission, toluidine blue dye lots must have at least 50% dye content. Unless corrected each time, toluidine blue solutions will vary in dye content.
Gary Gill
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-pi.csiro.au] Sent: Friday, July 06, 2001 5:04 AM To: microscopy-at-sparc5.microscopy.com
Continuing on this thread, I had always thought Richardson's stain was the methylene blue/azure II combination, but not so, it seems, since the 1960 paper refers to both this stain and to tol blue. Someone emailed that methylene blue must be used with azure II, but I don't think this is necessary, though it may give better staining.
Re metachromicity, as Fred Monson and Jan Coetzee note, it depends quite a bit on pH of the tol blue (for plant material at least), and used to depend on the source of stain but I guess this is not such a problem these days(?). We generally use a benzoate buffered solution of tol blue at pH 4.5 for fresh tissue and tissue embedded in hydrophilic resins like LR White and other methacrylates, and a pH 10 solution (buffered, if necessary, in sodium carbonate) for sections of hydrophobic epoxy resins - e.g. Spurr's. Tol blue metachromicity isn't so good at high pH (at least in my hands). The pH of tap water is usually slightly acidic, so aqueous tol blue is generally OK for fresh tissue and methacrylate resins. Since borate buffers are alkaline, perhaps this was the original reason they were used for the hydrophobic resins(?) - and they do produce a much more permanent stain.
Re. "old" references and books, most people here do molecular/genetic type work and literature older than about 1995 is considered venerable..... I am amazed (and appalled) at the great old histochemistry and cytochemistry books thrown out by libraries - the source of most of my "old" books.
Friday night, burning a few CDs..... Looked up another reference on histochemistry - A.G. Everson Pearse (1968) Histochemistry Vol 1 3rd edn (SBN 7000-1326-1 for Fred Monson - perhaps ISBN came after this time....) - a method from 1947 is given that uses a solution of 0.25% tol blue in 0.25% borax followed by 0.5% potassium dichromate in sat'd mercuric chloride, for permanent staining of paraffin sections prior to dehydration and mounting in Canada Balsam. Hess M, Hollander F (1947) J Lab Clin Med 32:905
Mallory, whose name used to be associated with various staining protocols, put it all together in a book: Mallory FB (1938) Pathological Technique. WB Saunders, Philadelphia. This is where Mallory's borax methylene blue comes from - 1% methylene blue plus 1% azure II in 1% borax. Lillie refers to it in his book - Histopathologic Technic - we have the 2nd edition, published in 1954, LC number 53-5622 - again, pre-ISBN.
Will finish now, in case this is getting too bor-ing!
cheers, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Yes geometry, accelerating voltage, sample composition and thickness have a huge effect, but you have to compare things correctly. Start with the detector and its window, if it isn't configured properly to start with then the sample and operating conditions can be irrelevant.
There are different windows out there, and not all windows are identical even if they are made from the same nominal material. Variation in thickness of the different components can make a significant difference in the performance of a window. This was shown in a round robin test run by Larry Thomas a long while back
Comparison of UTW/WL Detectors.... L.E. Thomas etal Analytical Electron Microscopy - 1984 San Francisco Press. pge 333-336.
It is true that the windows are more uniform today than they were back then, but there is still variation. In addition any contamination deposited on the window by the instrument environment (or ice formation inside the cryostat), and of course electronic noise will significantly deteriorate a detectors low energy performance.
If you wish to see a plot of detection efficiency vs detector window type (which DOES make a difference if the windows are different) lookup the following reference
N.J. Zaluzec , Evolutionary Developments in X-ray and Electron Energy Loss Microanalysis Instrumentation .... MicroBeam Analysis, 1990 San Francisco Press. pge 330-333.
Some of the figures from these two papers can also be found in this PDF documnet
Hello, My protein sample does not adsorb to the grid even after glow discharg, it is a membrane protein complex with the membrane lipids froming vescicles. Nothing show up after negative staining. What should I do to improve the adsorption? I have tried quick dring with filter paper after put the sample on the grid, but it does not work.
perfect microscope for home or garage or for any small lab needind an addition al imaging microscope. This scope is in great shape and needs a home with someone who can appreciate the fantastic imaging capabilities and the user serviceable nature of the scope!
Please check the Central Michigan University Purchasing Department Web site for details: http://www.purchasing.cmich.edu/frametemp/sales_main.html (go to the sealed bid items under "Pop Stops" or check the link below)
Nestor Zaluzec has a PPT presentation about EDS on the Argonne National Lab website that he advertised a few weeks back.
Part of that presentation deals with sensitivities for the various window materials and detector types. There are significant differences. No one detector is best for all situations and all elements. You may have to determine which detector is best for your combination of elements.
There should be similar critiques available through any number of sources, but they will probably have to be a general discussion. Manufacturers have a way of changing things and there may be something new, or at least slightly different on the market this week.
Warren
At 06:46 PM 7/6/2001 -0400, you wrote:
} Does anybody know of a good article that compares EDS detectors, from a } variety of manufacturers, w/ respect to their light element sensitivity? } } SMARTech } 860-491-3299 } www.semguy.com } 19 Cornwall Drive } Goshen CT 06756
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Hello, My protein sample does not adsorb to the grid even after glow discharg, it is a membrane protein complex with the membrane lipids froming vescicles. Nothing show up after negative staining. What should I do to improve the adsorption? I have tried quick dring with filter paper after put the sample on the grid, but it does not work.
Dear Mike, Unfortunately, the appropriate conditions for optimal coverage vary from sample to sample. You can change the hydrophobicity/hydrophyllicity of a thin carbon layer by "aging" over a few days for more hydrophobic carbon, glow discharging for more hydrophyllic carbon, and glow discharging in the presence of a volatile amine (we have amyl amine) for even more hydrophyllicity. You can also use either the side of the thin carbon--if, indeed, that is your substrate--which was in contact with mica--if the carbon was vacuum-evaporated onto freshly-cleaved mica--or the side away from the mica. Each of these has somewhat different properties. If you are using carbon-coated formvar as a substrate, you may still be able to alter its properties, but not in as many ways as with a thin carbon film. You do not say whether you are testing a preparation for cryo-EM by looking at a negatively-stained grid or whether you just want to look at a neg-st specimen. If the former, try blotting from one side only. Blotting from the back should leave more of the vesicles on the grid (although the thickness of the specimen when frozen can be hard to control). Also, you need to assure yourself that you can identify your specimen after mock cryo preparation followed by warming up and staining; it may look very little like a typical neg-st prep. If the latter, try letting the specimen with the stain sit on the grid for several minutes before blotting. This kind of preparation can be very tricky, and you just need to try different combinations of conditions until you get good coverage. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I guess that my original post was too curt. We have gone away from Tol Blue, to methylene blue due to safety reasons. There is no toxicology data on the MSDS for Tol Blue, but there is for Meth Blue. As I have a pregnant technician, I feel better knowing the LD 50 information, which is on the MSDS for Meth Blue. But, methylene blue is not as good for a general stain as Tol Blue, so I was just curious if there was an overriding reason to use it. I am presently experimenting with food based stains, which eliminates the issue completely.
The issue of pregnant (or possibly pregnant) workers in the lab has turned out to be fairly complicated. If anyone else out there has beem through this situation, I would like to know how you dealt with it.
Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] Sent: Thursday, July 05, 2001 10:20 AM To: 'Sally Stowe' Cc: 'Microscopy Listserver'
Dear Sally, To address the REAL attribution issue, one of us is going to have to go to a REALLY good library that has these OLD journals back to the late '50's. We may have to divide the labor, but the question is sufficiently interesting and meritorious that we ALL should get it right, when asked. Someone was responsible for the use of borax (Sodium tetraborate) with Toluidine blue O (1% in 1%). Here are all the citations that Pease provides and his [attributions]:
1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc Res., 5: 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in Lewis and Knight below)] 2. Charles and Meek , G.R. separately in a discussion of paper by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O in 1% borax used at 85 degrees C] 3. Pease, D.C.(1964), Histological Techniques for Electron Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 in 2st paragraph the exact method that I use for staining as 'his' method - "...flood the slide, cook until [edge] of drop [froths/]steams [or dries]....". [This is a long discussion to which Pease devotes an entire section, but one can just FEEL his impatience with the unimportance to HIM of this issue. The book, after all, is NOT about light microscopy. The semi-thin section is for orientation only, and hardly deserves any more notice - especially if the light microscopists cannot SEE what is to be seen in the best material ever handed them.] 4. In Kay, D.H.(ed)(1965), Techniques for electron Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this book pre-historic?!] The following is cited as THE reference for using Toluidine blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. cytol, 5: 508.
I have just looked in Geoffrey Meek's book (1970), Practical Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. 456-457. ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he has purposely NOT used citations at all.
I have also consulted the Sorvall manual on "Thin Sectioning"(3d Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated Technics for Electron Microscopy"(1959) which apparently was distributed with the MT-1. In the latter, only staining of Methacrylate sections was given and no mention of 1% *bor* of any kind. In the former, 1% borate at pH 4.5 is mentioned, but that is not relevant to this discussion.
I would bet that you might have some of these references. If you do not, let me know, and I will dig them up, in old Philadelphia, in Ben Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I will do it anyway, because I want to know, but I will do it quicker if you tell me I'm keeping you waiting.
Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Sally Stowe } Sent: Tuesday, July 3, 2001 3:37 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Tol blue/borax } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Somebody just asked me the reference for toluidine blue/sodium borate (1% } each) which is our routine LM stain - I cant find an attribution in any of } about 10 books. Can anybody help? This person should not be forgotten! } One suggestion was Alsop? } } thanks } } Sally } } } } Dr Sally Stowe } Facility Coordinator, } ANU Electron Microscopy Unit } Research School of Biological Sciences } Australian National University } Canberra ACT0200 } AUSTRALIA } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } ph 02 6125 2743 } http://www.anu.edu.au/EMU } } } } }
Woody, I certainly agree with you, but it seems Elaine was talking about qualitative or even quantitative comparison of the specimens histograms (I hope the specimens are flat). I think, the most straightforward thing is to use one of the measured specimens with wide brightness and contrast range. It makes no sense to calibrate BSE image for range from Si to Pb just to look at specimens containing, say, Cu and Au. The same for SE image. Polished specimen has much narrower signal range for brightness and contrast than fractured specimen. And, of course, for a microscope with a tungsten gun it is better to work well in the saturation range to insure brightness stability.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-mcdermott.com] } Sent: Monday, July 09, 2001 7:19 AM } To: 'Schumacher, Elaine'; Microscopy-at-sparc5.microscopy.com } Subject: RE: Brightness/Contrast Calibration Standards } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Elaine, } } A BSE standard can be made from almost any materials } (suitable for SEM). } The std should be ground and polished to remove topography } contrast. What } materials you choose will depend on the examination goals. } } This brings me to a point. I would not stress BSE "calibration" as a } generally useful technique. Under VERY limited circumstances, such } calibration/standardization can be beneficial. Routine use of this } technique will guarantee that sooner or later an important } phase will be } missed when examining an unknown. When examining with BSE, I find it } imperative to be "all over the map" with the BSE B&C to be } sure I am not } missing something. It is not unheard of to need two or more images to } define what is happening. Sometimes there is simply not } enough dynamic } range presented in one image to perceive all variations in } composition. } } Regards, } Woody White, McDermott Technology, Inc. } } } -----Original Message----- } } From: Schumacher, Elaine [mailto:efschuma-at-uop.com] } } Sent: Friday, July 06, 2001 12:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Brightness/Contrast Calibration Standards } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } } ---------. } } } } } } Fellow Microscopists, } } } } We're looking for suggestions for SEM standards to calibrate } } brightness/contrast settings (for example, setting an auto } } B/C feature in } } the SEM software, so that the desired repeatable B/C level is } } obtained by } } multiple users). Our customers' goal is to do } } correction/matching of gray } } scale histograms between images of different samples, and } } they would like to } } make the process of adjusting B/C levels at the start of } } image acquisition } } to be as standardized as possible. The imaging might be done } } in secondary } } or backscattered mode, so suggestions for both would be appreciated. } } } } Thanks vey much for you input, } } } } Elaine } } } } Elaine F. Schumacher } } Analytical Specialist } } UOP LLC } } 25 East Algonquin Road } } Des Plaines, IL 60017-5017 } } Phone: 847-391-3403 } } Fax: 847-391-3719 } } Email: efschuma-at-uop.com } } } } } }
Sputter deposition of Cu on the back side of holed samples was a common problem in one-sided thinning with the PIPS. The Cu came from the sample support posts that the sample was waxed onto. As Richard Beanland suggested, a simple acid cleaning (e.g., dilute nitric acid) removes the Cu effectively if the sample can take this treatment. Alternatively, you could vapor deposit a thin coating of rocksalt on the sample and later clean it off with water. Two-sided milling will avoid gross sputter contamination, but--
As a general rule in ion milling, it's highly recommended to avoid using any material in the sample fabrication or holder that you can't tolerate having in your sample. Whether you use a Cu or Mo grid, or fabricate samples by sandwiching the sample between Si blocks, for example, the nature of the ion milling process just about guarantees some of this material will collect in the sample cracks and crevices.
Larry Thomas Pacific Northwest National Laboratory, Richland, WA, USA larry.thomas-at-pnl.gov
--------- From: Ronald Anderson Sent: Monday, July 9, 2001 6:45 AM To: richard.beanland-at-marconi.com Cc: Feng Wu; Microscopy Listserver Subject: Re: GaN/Sapphire thinned by PIPS
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
The main tricks are to low-angle mill both sides of the specimen and to not use Cu grids. We switched to Mo grids years ago and we rarely see back sputtered material on samples milled on both sides (from the substrate direction, of course). If you want to use up your stock of Cu grids, cut the back side of the grid away so that there is no grid material beyond the specimen to sputter from. Grid becomes the "C" shaped grid used for FIB prepped specimens. Sometimes some sputtered material persists after very low angle milling. In that case, pop the sample into a conventional high-angle mill and mill at 25 degrees for 30 seconds at 2 or 3 KeV to clean off the sputter artifact.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Richard Beanland +44 1327 356363 {richard.beanland-at-marconi.com} on 07/02/2001 07:02:41 AM
To: Feng Wu {fwu-at-bgumail.bgu.ac.il} cc: Microscopy Listserver {microscopy-at-sparc5.microscopy.com} Subject: Re: GaN/Sapphire thinned by PIPS
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Feng, the trick is not to have a hole in your specimen where the ions can go through and back-sputter the Cu on to your sample. So make your specimen big enough to cover up the hole in the support grid and stop ion milling immediately when a hole appears. The problem will go away to some extent if you use the two-sided specimen holder. Alternatively, use an inert grid and dissolve the Cu with an acid after specimen prep. (Most acids are unlikely to affect GaN or sapphire.)
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } }
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Hello, My protein sample does not adsorb to the grid even after glow discharg, it is a membrane protein complex with the membrane lipids froming vescicles. Nothing show up after negative staining. What should I do to improve the adsorption? I have tried quick dring with filter paper after put the sample on the grid, but it does not work.
Thank you ......was written by a student with good intent and fair grammar/spelling. I apologize, lesson learned. Thanks for the help.
We as microscopists often receive requests for information about specific chemicals or chemical derivatives which we are requested to investigate or required to use in studies that are requested. The amount of information available is staggering, but relevant information is not always immediately available, and, for many of us, the means of access is not always apparent. The one datum that will provide you with the quickest access to information is the Chemical Abstracts Registry Number(CAS Reg. or just CAS). It is usually located in the catalog from which a chemical was ordered and can be obtained from the supplier and used to search any chemical database with specificity and speed. What follows is what I believe to be a reasonable strategy for free, rapid, and reasonable access to information about any chemical that raises a question. I have limited the list to four, because I have found what I need using no more. There are other sources, but my object was speed and utility.
A. NIST Chemistry WebBook(ver. 2000): http://webbook.nist.gov/
B. ToxLine Hazardous Substance Database: http://toxnet.nlm.nih.gov/cgi-bin/sis/htmlgen?HSDB
C. INTERNATIONAL CHEMICAL
SAFETY CARDS (ICSCs) (NIOSH/WHO): http://www.cdc.gov/niosh/ipcs/nicstart.html (US National Version)
D. Material Safety Data Sheets: http://siri.org/msds/index.html (This is the only site that is NOT supported by a government agency!)
For masters and students, I believe that the three URL's listed above provide access to the best information about chemicals that are considered hazardous.
DISCLAIMER: To assume that any chemical is harmless is foolish and probably wrong. Many years ago there was a news report of a pre-adolescent who woke up on 1November with diabetic acidosis after eating a grocery bag full of candy collected the night before. Even NaCl or KCl can be toxic!!! The Moral Is: If you get no return from any of the four above URL's for a chemical name with which you are NOT familiar, check the name and require a copy of the MSDS (Material Safety Data Sheet) BEFORE you permit it in your lab.
To date, I have used Google for over a year to search for info on everything including many organic chemicals. I have always received returns on chemical searches.
Respectfully submitted,
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
Below is the result of your feedback form. It was submitted by (drcline-at-flash.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, July 9, 2001 at 17:07:43 ---------------------------------------------------------------------------
Email: drcline-at-flash.net Name: Diane Cline
Organization: George Washington University
Education: Graduate College
Location: Washington DC
Question: My grandmother, Mrs. Ethel Schwartz, is trying to find an appropriate museum or library to donate about 500 slides of my grandfather's, Dr. J.B. Robbins, who developed the technique of photography through a triocular microscope of chemical crystals on slides in the late 196o's through early 1970's. Might you be able to suggest a collection (museum, university archive, or Library) that might be especially interested? Thank you, Sincerely Diane Harris Cline
I am interested in counting the total number of glomeruli in the kidneys in } rats with obstructive uropathy. To perform stereology, I have to cut thick } sections at 20 micrometers. I found that the kidney itself breaks down and } get very damaged at this thickness (kidneys fixed in 10% formalin and } embedded in parrafin), probably because the kidney is much harder than the } paraffin. In the litterature, some people embed kidneys in glycol } methacrylate. It might be better (harder), but then I do not know how to cut } my sections which are about 2.5 to 3 cm long, if I need a glass/diamond knife } that long. Any ideas about cutting thick sections in paraffin?, or is it } worth re-embedding in glycolmethacrylate, and any ideas about cutting my } serial sections at 20 micrometers in that resin? } } Thank you a lot for your help! } } F. Cachat } University of Virginia } fc6b-at-virginia.edu
Bill - here is the animal toxicity data according to the MSDS sheets available from our site. Methylene Blue: Oral rat LD50: 1180mg/kg Mutation references cited. Animal reproductive effects cited Toluidine Blue O: IPR-Rat LD50: 215mg/kg; IVN-Rat LD50: 28930?g/kg; IVN-Mus LD50: 45mg/kg; UNR-Mus LD50: 28mg/kg; IVN-Rbt LD50: 13440?g/kg. In an ideal world that data would be comparable, but this is what we have and I assume that its based on some realities. I would not overly worry about the LD50 figures, unless they indicate great toxicity. More concern is warranted if a toxin is cumulative and if its carcinogenic or mutagenic. Mutagenicity is indicated for methylene blue and not for toluidine blue O. Since mutagenicity is one of the great fears during pregnancy, I would suggest to revert to toluidine blue O. I think that most people in this forum would understand that carcinogenic and mutagenic molecules are ingested daily and in huge numbers. Clearly the body wards off all of these and it is a very rare event that a cancer actually forms. It is well known that a person with a healthy life-style and on a good diet (particularly one rich in the anti-oxidant vitamins) is more likely to avoid cancer. That healthy life-style includes minimizing, but never excluding contact with "adverse" substances. I suggest that one should be circumspect, but avoid paranoia. On balance, I regard a well run EM lab as one of the safer workplaces about; it would be nice to have comparative data of sick days or better still accident data, but I doubt that they exist. While on the topic of safety and carcinogens I like to tell a little story. It happened at an unmentionable university about 15 years ago. The ceiling in one large public room was found to contain fibres, which were confirmed to be asbestos. After some agitation the university's response was to invite an "expert", who was used by various organisations to "resolve" asbestos problems. The expert soon had his first meeting with union officials during which he insisted that asbestos was not very dangerous. To make his point, he opened a jar allegedly containing powdered asbestos and threw a handful into the air. Even 15 years ago, such action identified that man as an idiot. Educated perhaps, but an idiot nonetheless. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, July 10, 2001 5:05 AM, William McManus [SMTP:billemac-at-biology.usu.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To Sally and all: } } I guess that my original post was too curt. We have gone away from Tol } Blue, to methylene blue due to safety reasons. There is no toxicology } data on the MSDS for Tol Blue, but there is for Meth Blue. As I have a } pregnant technician, I feel better knowing the LD 50 information, which } is on the MSDS for Meth Blue. But, methylene blue is not as good for a } general stain as Tol Blue, so I was just curious if there was an } overriding reason to use it. I am presently experimenting with food } based stains, which eliminates the issue completely. } } The issue of pregnant (or possibly pregnant) workers in the lab has } turned out to be fairly complicated. If anyone else out there has beem } through this situation, I would like to know how you dealt with it. } } Bill } } William McManus } Supervisor } Electron Microscopy Facility } Department of Biology } Utah State University } Logan UT 84322-5305 } } billEMac-at-cc.usu.edu } 435-797-1920 } } -----Original Message----- } } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] } Sent: Thursday, July 05, 2001 10:20 AM } To: 'Sally Stowe' } Cc: 'Microscopy Listserver' } Subject: RE: Tol blue/borax } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Sally, } To address the REAL attribution issue, one of us is going to } have to } go to a REALLY good library that has these OLD journals back to the } late } '50's. We may have to divide the labor, but the question is } sufficiently } interesting and meritorious that we ALL should get it right, when asked. } Someone was responsible for the use of borax (Sodium tetraborate) with } Toluidine blue O (1% in 1%). Here are all the citations that Pease } provides } and his [attributions]: } } 1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc } Res., 5: } 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in } Lewis } and Knight below)] } 2. Charles and Meek , G.R. separately in a discussion of } paper } by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O } in } 1% borax used at 85 degrees C] } 3. Pease, D.C.(1964), Histological Techniques for Electron } Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 } in } 2st paragraph the exact method that I use for staining as 'his' method - } "...flood the slide, cook until [edge] of drop [froths/]steams [or } dries]....". [This is a long discussion to which Pease devotes an } entire } section, but one can just FEEL his impatience with the unimportance to } HIM } of this issue. The book, after all, is NOT about light microscopy. The } semi-thin section is for orientation only, and hardly deserves any more } notice - especially if the light microscopists cannot SEE what is to be } seen } in the best material ever handed them.] } 4. In Kay, D.H.(ed)(1965), Techniques for electron } Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this } book } pre-historic?!] The following is cited as THE reference for using } Toluidine } blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. } cytol, 5: 508. } } I have just looked in Geoffrey Meek's book (1970), Practical } Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. } 456-457. } ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he } has } purposely NOT used citations at all. } } I have also consulted the Sorvall manual on "Thin Sectioning"(3d } Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated } Technics for Electron Microscopy"(1959) which apparently was distributed } with the MT-1. In the latter, only staining of Methacrylate sections } was } given and no mention of 1% *bor* of any kind. In the former, 1% borate } at } pH 4.5 is mentioned, but that is not relevant to this discussion. } } I would bet that you might have some of these references. If } you do } not, let me know, and I will dig them up, in old Philadelphia, in Ben } Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I } will } do it anyway, because I want to know, but I will do it quicker if you } tell } me I'm keeping you waiting. } } Regards, } } Fred Monson } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI) } Schmucker II Science Center (Room: SS024(Basement)) } South Church Street } West Chester, PA, 19383 } MailDrop: Department of Geology/Astronomy } Phone: 610-738-0437 } Fax: 610-436-3036 } email: fmonson-at-wcupa.edu } Please call before visiting. } } } } } } ---------- } } From: Sally Stowe } } Sent: Tuesday, July 3, 2001 3:37 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Tol blue/borax } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } Somebody just asked me the reference for toluidine blue/sodium borate } (1% } } each) which is our routine LM stain - I cant find an attribution in } any of } } about 10 books. Can anybody help? This person should not be } forgotten! } } One suggestion was Alsop? } } } } thanks } } } } Sally } } } } } } } } Dr Sally Stowe } } Facility Coordinator, } } ANU Electron Microscopy Unit } } Research School of Biological Sciences } } Australian National University } } Canberra ACT0200 } } AUSTRALIA } } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } } ph 02 6125 2743 } } http://www.anu.edu.au/EMU } } } } } } } } } }
Hi Jan, Just to keep the historical stuff together on this thread, the well-known Giemsa blood stain was first published, as a footnote, in "...an obscure journal...." in (see Gray, P.(1954), The Microtomist's Formulary and Guide, Blakiston, NY,NY. (reprinted by Krieger(1975), ISBN 0-88275-247-2). The dry Giemsa stain is an eosinate of methylene blue [acid + base = insoluble salt (in HOH but not in MeOH! Later, on the slide, soluble (mass action) in large excess of MeOH-water or buffer mix yields differential dyeing].
I searched the Krieger site (http://www.web4u.com/krieger-publishing/) for ISBN and came up with the following:
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THE MICROTOMIST'S FORMULARY AND GUIDE Peter Gray, , 0-88275-247-2
Pages: 808, Binding: Cloth, Price: $81.50
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This is a known and recognized source reference work. The book includes a treatise on the art of making microscopic slides from biological specimens, as well as a classified list of the formulas and techniques used in this art.
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Orig. Ed 1954, Reprint Ed. 1975
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Fred Monson
} ---------- } From: Jan Coetzee } Reply To: janc-at-ccnet.up.ac.za } Sent: Friday, July 6, 2001 3:55 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Tol blue/borax } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } } To contribute to the search for the roots of the Toluidine blue } procedure: } } There is an interesting paper by Kramer & Windrum (J. Histochem } Cytochem, 1955, Vol 3, 227-237, The Metachromic Staining Reaction) } that probably was instrumental in initiating the wider use of Tol } Blue. In this paper the effect of stain concentration and residual } water is shown (amongst many other factors) to be important for the } development of the multiplicity of colours. } } The use of Toluidine Blue as a stain for monitor sections of epoxy- } and acrylic-embedded tissue probably stems from these metachromic } effects that this stain produces, with the result that some deductions } regarding the chemical composition of the sample can be made after a } simple Tol Blue staining. } } Even though we, in this lab, have since forever used Tol Blue for the } staining of monitor sections of resin-embedded tissues, every batch of } Tol Blue that we make up seems to have slightly different staining } characteristics. This was such a source of irritation that Chris van } der Merwe recently decided to systematically investigate some of the } variables. } } He found that the most important parameters are: } 1. Tol blue concentration (0.1 or 0.5%) } 2. Buffer or vehicle type, pH and concentration (0.1 - 1% } Na-carbonate, Na-benzoate buffer at pH4, Na-tetraborate (borax) at pH } 8.8) } 3. Embedding resin formulation (Spurr, Quetol, Polarbed 812, LR-White) } 4. Section thickness. } } All the above have an effect on the intensity, as well as the degree } of metachromicity, of the stain. } Resin formulation is probably the major influencer of the results. } } This may be a reason for the existence of the wide variety of Tol Blue } recipes that abound in the literature. } } More information on Tol Blue staining can be had directly from Chris } at cvdmerwe-at-scienti.up.ac.za } } } Regards } } Jan Coetzee } Lab for Microscopy and Microanalysis } University of Pretoria, South Africa. } Tel: 012-420-2075, Fax 012-362-5150 } www.up.ac.za/academic/electron/emunit1.htm } } } } "Monson, Frederick C." wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Sally, } } To address the REAL attribution issue, one of us is going to } have to } } go to a REALLY good library that has these OLD journals back to the } late } } '50's. We may have to divide the labor, but the question is } sufficiently } } interesting and meritorious that we ALL should get it right, when asked. } } Someone was responsible for the use of borax (Sodium tetraborate) with } } Toluidine blue O (1% in 1%). Here are all the citations that Pease } provides } } and his [attributions]: } } } } 1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc } Res., 5: } } 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in } Lewis } } and Knight below)] } } 2. Charles and Meek , G.R. separately in a discussion of } paper } } by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O } in } } 1% borax used at 85 degrees C] } } 3. Pease, D.C.(1964), Histological Techniques for Electron } } Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 } in } } 2st paragraph the exact method that I use for staining as 'his' method - } } "...flood the slide, cook until [edge] of drop [froths/]steams [or } } dries]....". [This is a long discussion to which Pease devotes an } entire } } section, but one can just FEEL his impatience with the unimportance to } HIM } } of this issue. The book, after all, is NOT about light microscopy. The } } semi-thin section is for orientation only, and hardly deserves any more } } notice - especially if the light microscopists cannot SEE what is to be } seen } } in the best material ever handed them.] } } 4. In Kay, D.H.(ed)(1965), Techniques for electron } } Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this } book } } pre-historic?!] The following is cited as THE reference for using } Toluidine } } blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. } } cytol, 5: 508. } } } } I have just looked in Geoffrey Meek's book (1970), Practical } } Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. } 456-457. } } ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he } has } } purposely NOT used citations at all. } } } } I have also consulted the Sorvall manual on "Thin Sectioning"(3d } } Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated } } Technics for Electron Microscopy"(1959) which apparently was distributed } } with the MT-1. In the latter, only staining of Methacrylate sections } was } } given and no mention of 1% *bor* of any kind. In the former, 1% borate } at } } pH 4.5 is mentioned, but that is not relevant to this discussion. } } } } I would bet that you might have some of these references. If } you do } } not, let me know, and I will dig them up, in old Philadelphia, in Ben } } Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I } will } } do it anyway, because I want to know, but I will do it quicker if you } tell } } me I'm keeping you waiting. } } } } Regards, } } } } Fred Monson } } } } Frederick C. Monson, PhD } } West Chester University of Pennsylvania } } Center for Advanced Scientific Imaging (CASI) } } Schmucker II Science Center (Room: SS024(Basement)) } } South Church Street } } West Chester, PA, 19383 } } MailDrop: Department of Geology/Astronomy } } Phone: 610-738-0437 } } Fax: 610-436-3036 } } email: fmonson-at-wcupa.edu } } Please call before visiting. } } } } } } } ---------- } } } From: Sally Stowe } } } Sent: Tuesday, July 3, 2001 3:37 AM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: Tol blue/borax } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Somebody just asked me the reference for toluidine blue/sodium borate } (1% } } } each) which is our routine LM stain - I cant find an attribution in } any of } } } about 10 books. Can anybody help? This person should not be } forgotten! } } } One suggestion was Alsop? } } } } } } thanks } } } } } } Sally } } } } } } } } } } } } Dr Sally Stowe } } } Facility Coordinator, } } } ANU Electron Microscopy Unit } } } Research School of Biological Sciences } } } Australian National University } } } Canberra ACT0200 } } } AUSTRALIA } } } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } } } ph 02 6125 2743 } } } http://www.anu.edu.au/EMU } } } } } } } } } } } } } } } } }
You may have better luck using a vibratome. The kidney, being sharply demarcated and well encapsulated, lends itself to processing in this manner. I would fix before sectioning, (dependent on species donating the kidney-probably not bigger than rat) but I have seen it work either way. The fixative is dictated by stains/probes of interest. Using a cryotome and a water soluble embedding media has also worked well for me in the past. good luck
Ramin
-----Original Message----- } From: "CachatF-at-aol.com"-at-sparc5.microscopy.com [mailto:"CachatF-at-aol.com"-at-sparc5.microscopy.com] Sent: Monday, July 09, 2001 11:19 PM To: Microscopy-at-sparc5.microscopy.com
I am interested in counting the total number of glomeruli in the kidneys in } rats with obstructive uropathy. To perform stereology, I have to cut thick
} sections at 20 micrometers. I found that the kidney itself breaks down and
} get very damaged at this thickness (kidneys fixed in 10% formalin and } embedded in parrafin), probably because the kidney is much harder than the
} paraffin. In the litterature, some people embed kidneys in glycol } methacrylate. It might be better (harder), but then I do not know how to cut } my sections which are about 2.5 to 3 cm long, if I need a glass/diamond knife } that long. Any ideas about cutting thick sections in paraffin?, or is it } worth re-embedding in glycolmethacrylate, and any ideas about cutting my } serial sections at 20 micrometers in that resin? } } Thank you a lot for your help! } } F. Cachat } University of Virginia } fc6b-at-virginia.edu
At 1:04 PM -0600 7/9/01, William McManus wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When I was pregnant, 9 years ago, I was the entire staff of the EM facility here (still was until a month ago)and so could not avoid things. The way I handled safety concerns was to be extra cautious: I worked in the hood, wore double gloves when working with fixatives & resins and gloves for other times when i would not usually wear them( like using Tol Blue), safety glasses if splashing was a possibility, washed my hands almost compulsively. In short, I used common sense. I was, and am, lucky that (UrAc aside) I don't have to deal with radioactivity. colleagues of mine who were also pregnant at the time (we had a rash of pregnancies that year) who did need to work with isotopes minimized their use, got new shields, and at times convinced labmates to do those steps for them!
We all, thankfully had healthy babies who are now all approaching adolescence and driving us nuts, but that's another issue for another forum!
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} I am interested in counting the total number of glomeruli in the kidneys in } } rats with obstructive uropathy. To perform stereology, I have to cut thick } } sections at 20 micrometers. I found that the kidney itself breaks down and } } get very damaged at this thickness (kidneys fixed in 10% formalin and } } embedded in parrafin), probably because the kidney is much harder than the } } paraffin. In the litterature, some people embed kidneys in glycol } } methacrylate. It might be better (harder), but then I do not know how to } cut } } my sections which are about 2.5 to 3 cm long, if I need a glass/diamond } knife } } that long. Any ideas about cutting thick sections in paraffin?, or is it } } worth re-embedding in glycolmethacrylate, and any ideas about cutting my } } serial sections at 20 micrometers in that resin? } } } } Thank you a lot for your help! } } } } F. Cachat } } University of Virginia } } fc6b-at-virginia.edu
How about frozen sections? Fix in formalin, cryoprotect with ascending concentrations of sucrose (10%, 20%, 30%) then snap freeze and section at 20 microns on a sliding microtome. Collect sections in ice-cube trays to keep them in order. In my experience, thick paraffin sections are much more difficult than thinner ones. Embedding tissue in methacrylates (and other plastics) is usually intended for thinner sections (1-3 microns).
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
We read your message with great interest.i routinely fix kidney of approximately the same size but usually cut for stereological work -at-10u. you do not state any info regarding processing times etc but if your tissue is brittle this could be due to numerous factors over fixation leaving in alcohol/clearing agent too long (especially chloroform as clearing agent) ( i come from a laboratory that hand processes, which gives more control over times processed compared to that from machines.) i also would question the fixative i always use Bouins fixative with kidney and have never had any problems in sectioning. i would suggest that you do not embed in glycolmethacrylate as this is for cutting thin's and also there is no way you would be able to get diamond knife to cut the size if tissue you want to. you could make ralph knives but again this will probably not work on the thickness you are after.
my last suggestion to you is regarding you 'already brittle blocks' try bakers soaking fluid (mixture of alcohol/glycerine) this should cure your problem but it is slow going as you will only get a few sections before they will need resoaking
i hope this works and is helpful to you yours
Phil
Mr P R Mutch Senior Imaging/Research Technician Nottingham University School of Biomedical Sciences E Floor, Medical School Queens Medical Centre Clifton Boulevard Nottingham NG7 2UH UK Tel 0115 9709415 E-mail philip.mutch-at-nottingham.ac.uk
Mr P R Mutch Senior Imaging/Research Technician Nottingham University School of Biomedical Sciences E Floor, Medical School Queens Medical Centre Clifton Boulevard Nottingham NG7 2UH UK Tel 0115 9709415 E-mail philip.mutch-at-nottingham.ac.uk
Hello, my name is Chen Chen, in Johns Hopkins University School of Medicine. Mike DeLannoy introduced me to subscribe to this group. Thank you Chen Chen
This is what I have done so far: 1. Removed all stored chemicals from the lab. 2. Moved all chemical mixing into fume hood. 3. Met for 3 hours with environmental safety officer. 4. Met for 2 hours with radiation safety officer. 5. Eliminated all protocols which involve teratogens from pregnant workers routine. 6. Established a personal hygiene plan for pregnant worker, which is being signed of by worker, myself, director of the lab, safety officer, and her physician. 7. Reiterated safety protocols for worker, coat , gloves, safety glasses, etc. 8. Had lab monitored for radiation, passed. 9. Had lab monitored for formaldehyde gas, passed. 10. Planning to monitor xylene exposure to worker when thin sectioning (cannot do this in the hood, obviously). Heat pen( modified soldering iron) has not worked well so far, any ideas how to eliminate xylene? 11. Still looking for a non toxic general 1 micron stain for Spurrs and LR White. Any ideas? I am experimenting with food related stains (grape juice, etc.). 12. Employee has completed, lab safety training course, formaldehyde training course and radiation training course. 13. Will continue to monitor lab for chemical and radation exposure.
To some this may seem like over kill, but I believe that it is very important that every stone is turned in regards to safety. Another issue that I face is that I have numerous grad students coming through the lab, and there is always a possibility that they are pregnant and do not realize it or keep it from me. I also have a nursing mother who was visiting the lab and never bothered to tell me, until the subject of why the lab was reorganized came up. I now demand that all female student, techs, visitors, ect. tell me of their "motherhood" status.
Bill
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Tuesday, July 10, 2001 7:28 AM To: William McManus; Monson, Frederick C.; Sally Stowe Cc: Microscopy Listserver
Every summer at our institute we sponsor a summer camp program called Adventures in Science. We invite inner-city fifth graders to come and hopefully see how much fun science can be. It has been rather successful and we all seem to enjoy the week.
One of the scientists here wants to give a demonstration involving vortices and fluids. He has a few short demonstrations to show the kids, but I told him I think he needs more to keep their interest and to keep them busy. He has five groups of 6 children for about an hour.
I told him about the MSA newsgroup and how helpful you all are and said I would post this question. I know this is not a microscopy question but I thought some of you might have some suggestions.
Thanks in advance for any help.
Peggy Bisher --
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
I commend you for your concern about safety. I went through two pregnancies as a biological electron microscopist. Since I was the solitary person doing TEM, I didn't have the choice of delegating anything. I actually chose to delay an R&D project for my first trimester because I didn't want to use uranyl acetate, osmium tetroxide/ruthenium tetroxide and especially lead citrate and tannic acid stains all involved in TEM. Other than that, I just used extra caution, especially when weighing/handling powdered chemicals. I never paid much attention to the Tol. blue - you have obviously looked into this in great detail.
I missed your original post. Are you trying to stain epoxy-embedded sections for light microscopy? I've tried a number of food dyes in the past and had very limited success. Also just because they are FD&C or D&C approved doesn't guarantee safety. I've worked with Erioglaucine (Brilliant Blue FCF, FD&C Blue #1) and the MSDS states it is a possible carcinogen.
I've tried some non-traditional dyes on epoxy sections of mammalian tissue. The standard Tol. Blue, "Epoxy Tissue Stain" (mixture with Basic Fuchsin) and/or Azur II always worked the best. BTW, my MSDS's listed Toluidine Blue as an irritant, but Methylene Blue as an irritant and a mutagen. Therefore I would think Tol. Blue was a little safer.
Excerpts from an experiment I did in '97 trying different stains on ~1micron sections of skin embedded in Spurr's resin (also not a good safety choice): Dyes tested: 0.1% aqueous Phloxine B. [related to eosin, a useful D&C approved dye for fresh tissue and fluorescence. ] 0.15% Sudan Black B in 70% ethanol. 0.5% aqueous Neutral Red. [a fairly nontoxic dye (although MSDS states possible mutagen) that we use a lot in fresh tissue.] Zymed Methyl Green. All were done on a hotplate for 30-60 seconds
Staining Results: Phloxine B seemed to stain only the dermal collagen with a fairly bright pink stain. Sudan Black B produced an interesting pattern of staining small elongated strips throughout the dermis, as well as intracellular staining of basal epidermal cells. The fat cell staining was non-uniform. Neutral Red stained in a very similar pattern to Sudan Black B in the dermis, only much more faint. Methyl Green produced a strong general stain similar but not exactly the same as Toluidine Blue.
Hope this is useful, Karen -------------------------------------------------------------------------------- Karen S. Zaruba BioMaterials Technology Center 3M Center Bldg. 270-1S-01 Tel: (651) 737-2971 Fax: (651) 737-5645
To: "Monson, Frederick C." {fmonson-at-wcupa.edu} "Sally Stowe" {STOWE-at-rsbs.anu.edu.au} cc: "Microscopy Listserver" {microscopy-at-sparc5.microscopy.com} (bcc: Karen S. Zaruba/US-Corporate/3M/US) Subject: RE: Tol blue/borax
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To Sally and all:
I guess that my original post was too curt. We have gone away from Tol Blue, to methylene blue due to safety reasons. There is no toxicology data on the MSDS for Tol Blue, but there is for Meth Blue. As I have a pregnant technician, I feel better knowing the LD 50 information, which is on the MSDS for Meth Blue. But, methylene blue is not as good for a general stain as Tol Blue, so I was just curious if there was an overriding reason to use it. I am presently experimenting with food based stains, which eliminates the issue completely.
The issue of pregnant (or possibly pregnant) workers in the lab has turned out to be fairly complicated. If anyone else out there has beem through this situation, I would like to know how you dealt with it.
Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
What I do is after they are hired, or at the beginning of the class, in front of witnesses, I explain that for their own safety that I must know if they are pregnant, so that all the appropriate precautions are taken. I also tell them that this information will be kept in strict confidence. It is pretty easy for people to figure out what is going on, however, once you start treating one person differently than the rest.
Bill
-----Original Message----- } From: Chuck Butterick [mailto:cbutte-at-ameripol.com] Sent: Tuesday, July 10, 2001 1:31 PM To: William McManus
This is what I have done so far: 1. Removed all stored chemicals from the lab. 2. Moved all chemical mixing into fume hood. 3. Met for 3 hours with environmental safety officer. 4. Met for 2 hours with radiation safety officer. 5. Eliminated all protocols which involve teratogens from pregnant workers routine. 6. Established a personal hygiene plan for pregnant worker, which is being signed of by worker, myself, director of the lab, safety
officer, and her physician. 7. Reiterated safety protocols for worker, coat , gloves, safety glasses, etc. 8. Had lab monitored for radiation, passed. 9. Had lab monitored for formaldehyde gas, passed. 10. Planning to monitor xylene exposure to worker when thin sectioning (cannot do this in the hood, obviously). Heat pen( modified soldering iron) has not worked well so far, any ideas how to eliminate xylene? 11. Still looking for a non toxic general 1 micron stain for Spurrs and LR White. Any ideas? I am experimenting with food related stains (grape juice, etc.). 12. Employee has completed, lab safety training course, formaldehyde training course and radiation training course. 13. Will continue to monitor lab for chemical and radation exposure.
To some this may seem like over kill, but I believe that it is very important that every stone is turned in regards to safety. Another issue that I face is that I have numerous grad students coming through the lab, and there is always a possibility that they are pregnant and do not realize it or keep it from me. I also have a nursing mother who was visiting the lab and never bothered to tell me, until the subject of why
the lab was reorganized came up. I now demand that all female student, techs, visitors, ect. tell me of their "motherhood" status.
Bill
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Tuesday, July 10, 2001 7:28 AM To: William McManus; Monson, Frederick C.; Sally Stowe Cc: Microscopy Listserver
At 1:04 PM -0600 7/9/01, William McManus wrote: } -----------------------------------------------------------------------
- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi,
I'm looking for anyone who may have images of red blood cells, white blood cells, and platelets. I'm also looking for an illustration of haemoglobin binding. My mother-in-law is working on an educational web page about blood for grades 4-6. All images will be referenced and properly credited. If you can help, I would greatly appreciate it.
Postdoctoral position(s): Dynamics of biological machines
Postdoctoral position(s) are available in the McGough lab at Purdue University to study biological machines by a variety of microscopic, computational, and biochemical approaches. Current projects in the lab include structure-function studies of the cytoskeleton and bacteriophage assembly.
Successful candidate(s) will work in close collaboration other laboratories at Purdue in cross-disciplinary studies using a variety of approaches including protein biochemistry, video light microscopy, time-resolved electron cryomicroscopy, atomic force microscopy, magnetic tweezers, and molecular modeling. Highly qualified candidates will be encouraged to develop new collaborations involving members of the cytoskeleton, structural biology, and single molecule groups at Purdue.
Applicants should have a Ph.D. in engineering or in one of the physical or life sciences. The ideal candidate will be a hard-working, enthusiastic individual with strong communication skills, experience in protein biophysics, and a commitment to applying an integrated structure-function approach to study biological machines.
Salary is negotiable.
Interested individuals should contact:
Dr. Amy McGough Department of Biological Sciences Purdue University West Lafayette, IN 47907-1392 USA phone: +1 (765) 496-7501 fax: +1 (765) 496-7501 amcgough-at-bilbo.bio.purdue.edu http://bilbo.bio.purdue.edu/~amcgough/
Purdue is an equal access/equal opportunity university.
} On the other hand, if newer technologies could improve your performance } significantly, then you might want to consider upgrading. The } downside? When the PC that is running the latest system becomes obsolete } (3 years? 5 years?) and the rest of your system (vacuum, High Voltage, } etc.) is still fine, what are you going to do? Try to find a new 486 with } Win 3.1 to replace a crashed but fairly recent Amray, for example. Their } boards and software won't run on anything newer. I'm not picking on them, } I'm just familiar with them.
I'm running an Amray 1910FE with PC control over the Nibblenet. The system came with a Dell 486 and 250MB hard drive. I am actually using a Pentium III/450 with Win95 or Win98 (either works fine). The 8741 MPU on the Nibblenet board in the PC needs the latest firmware version (2.3 or 2.4) to run with a Pentium. The board is hardwired to IRQ9. With a simple PC system, there are no IRQ conflicts.
I do not use the Image Control software/hardware (Matrox IP8) but rather use Soft Imaging ADDA II for active beam control and image capture and the GrabBit frame grabber to capture TV video. Console electronics are un-changed. PC-10 video goes to one of the GrabBit RS-190 inputs. The other input is from the chamber view camera.
So far, KLA/Amray is still supporting the FE systems.
} One of the scientists here wants to give a demonstration involving } vortices and fluids. He has a few short demonstrations to show the } kids, but I told him I think he needs more to keep their interest and } to keep them busy. He has five groups of 6 children for about an } hour.
Hi, Peggy --
I'll give you a great idea that takes about ten bucks worth of parts and a source of compressed air. How about a gizmo made of two 1/2-inch (OK, Ok, 12mm) pipes, a washer, and a ball valve which appears to contain Maxwell's demon. Blow compressed air in the middle and presto, hot air comes out one pipe and cold air comes out the other! It's called a Hilsch tube, and here's a link:
http://www.visi.com/~darus/hilsch/
If you do a search on "hilsch tube" you can find some commercial suppliers of ready-made units.
I remember seeing it in an old issue of Scientific American, the Amateur Scientist column. An outfit called Tinker's Guild, which I believe to be the creation of Shawn Carlson, current author of the Amateur Scientist, is offering a CD containing *every Amateur Scientist column written since 1928* for a snappy $70. Quite the bargain:
http://www.tinkersguild.com
Any of you who are science educators should get a copy of this! No commercial interest, just a fascinated customer who remembers actually trying quite a few of these as a kid.
We have a student who has been looking at biofilms on artificial membranes and in the process found a strange-looking inclusion. If there's anyone out there who has the time to have a look at this (it is displayed on http://www.ru.ac.za/emu/lechbug.htm) and give any suggestions this will be much appreciated.
Regards
Rob
======================================================= Robin H Cross (Mr) Director : Electron Microscopy Unit Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/index.htm
** remember ICEM-15 in Durban in September 2002 **
} From: "Rick Mott" {rickmott-at-alumni.princeton.edu} } } Peggy Bisher wrote: } } } One of the scientists here wants to give a demonstration involving } } vortices and fluids. He has a few short demonstrations to show the } } kids, but I told him I think he needs more to keep their interest and } } to keep them busy. He has five groups of 6 children for about an } } hour. } } Hi, Peggy -- } } I'll give you a great idea that takes about ten bucks worth of parts } and a source of compressed air. How about a gizmo made of two } 1/2-inch (OK, Ok, 12mm) pipes, a washer, and a ball valve which } appears to contain Maxwell's demon. Blow compressed air in } the middle and presto, hot air comes out one pipe and cold air } comes out the other! It's called a Hilsch tube, and here's a link: } } http://www.visi.com/~darus/hilsch/ } } If you do a search on "hilsch tube" you can find some commercial } suppliers of ready-made units. } } I remember seeing it in an old issue of Scientific American, the } Amateur Scientist column. An outfit called Tinker's Guild, which } I believe to be the creation of Shawn Carlson, current author of } the Amateur Scientist, is offering a CD containing *every Amateur } Scientist column written since 1928* for a snappy $70. Quite } the bargain: } } http://www.tinkersguild.com } } Any of you who are science educators should get a copy of } this! No commercial interest, just a fascinated customer who } remembers actually trying quite a few of these as a kid. } The Amateur Scientist is no more. It got cut along with most of the other columns. It looks like a very large short fall in advertising revenues are a problem. The adds are almost all consumer advertising. Not much that match the readership. With some of the politically correct stuff they have been publishing the last few years I expect that the scientific readership is being run off and it is over the lay readerships head. I was particularly put off by their piece on terrorism in the USA with out a any mention of environmental terrorism.
The Amateur Scientist got a great number of us started on the road to science it is a shame that it is being let go.
I have heard Fry's has the CD-ROM on sale for $25 with a $25 mail in rebate. Now that's a deal.
Cold Spring Harbor Laboratory is seeking an experienced and responsible microscopy technician for the laboratory's core biological microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, and digital imaging. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org -- David L. Spector Cold Spring Harbor Laboratory One Bungtown Road Cold Spring Harbor, New York 11724 Tel. (516) 367-8456 Fax (516) 367-8876 email: spector-at-cshl.org
} Dear Sir, } } I am a media artist based in Canada. I am presently working on a low } budget, art funded (Canada Council for the Arts) DVD project destined for } non-profit art gallery use. } } The work is a philosophic and poetic look at evolution and bio-diversity. } } I am very interested in getting and using some of the video your members may } have made on primitive life forms such as viruses, bacteria, etc. } I would be also interested in } acquiring from you a high quality beta-sp or preferable dv tape you might } have of these studies as well as other studies you may have of microscopic } life forms/elements dna, diatoms, etc. Full credit to your organization } (and the videographer if desired) would be given in the final piece. I } would pay for all shipping and handling costs. } } } } The video and images would be a small component of a much longer work. } } My last work recently screened at the Canadian Embassy in Washington, D.C. } and I have had shows at the Museum of Modern Art in New York City and at } many other prestigious venues around the world. I include my bio. below. } } Please get back to me regarding my request. } } } Best Wishes, } Oliver Hockenhull } } BIO - 2001 Oliver Hockenhull } } Home Site: http://www.crosswinds.net/~ideoplastic/ } (From this site link to various works in VRML, hypertext, etc.) } } Oliver Hockenhull is an award-winning media artist, director, communication } theorist, and lecturer. } } He has completed five feature length works and numerous dramatic and } experimental shorts. The films have shown internationally at such film } festivals and venues as -(selected) The Kerala-India International Film } Festival, The Montréal World Film Festival, The Leipzig International } Documentary Film Festival, The Vancouver International Film Festival, The } European Media Arts Festival, The Nouveau Cinema Festival , The } International Documentary Film Festival of Amsterdam, The Museum of Modern } Art in New York City, the National Gallery of Art in Washington, D.C., and } LACE (Los Angeles Contemporary Exhibitions).
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
A neat topic is eddy currents. There are a number of ways to experiment with them.
My son did a project that I helped him with. I first showed him the effect by getting a magnet and tying a string to it and used it as a pendulum. We counted the number of periods for it to stop. Then we got a an aluminum tray and put it under the swinging magnet and counted again. We changed the distance of the tray to see the effect.
Then we got fancy. We built a swinging pendulum in which we could put sheets of metals that we got from the hobby store. We used aluminum, brass, and titanium. The sheets could pass through magnets that we set up in epoxy on a board that fit onto the base of the pendulum with pegs so that they put in the same place every time. We made two sets of magnets with different gaps. The magnets were bought at Radio Shack. We experimented with different thickness of sheets, and stacks of sheets to make the same thickness. We experimented with sheets of paper that isolated the sheets of metal. It was really a nice little project that illustrated a number of effects.
To get even more fancy, you could do the same things with disks of metal rotating on an inexpensive dc motor. You could then monitor the current with a meter to measure the power change with different magnetic fields.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Peggy Bisher [mailto:peggy-at-research.nj.nec.com] Sent: Tuesday, July 10, 2001 12:07 PM To: Microscopy-at-sparc5.microscopy.com
Dear Microscopists,
Every summer at our institute we sponsor a summer camp program called Adventures in Science. We invite inner-city fifth graders to come and hopefully see how much fun science can be. It has been rather successful and we all seem to enjoy the week.
One of the scientists here wants to give a demonstration involving vortices and fluids. He has a few short demonstrations to show the kids, but I told him I think he needs more to keep their interest and to keep them busy. He has five groups of 6 children for about an hour.
I told him about the MSA newsgroup and how helpful you all are and said I would post this question. I know this is not a microscopy question but I thought some of you might have some suggestions.
Thanks in advance for any help.
Peggy Bisher --
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
} Sorry, forgot to include his email. } } } } Can anyone help this guy?? } } } } } Dear Sir, } } } } I am a media artist based in Canada. I am presently working on a low } } budget, art funded (Canada Council for the Arts) DVD project destined for } } non-profit art gallery use. } } } } The work is a philosophic and poetic look at evolution and bio-diversity. } } } } I am very interested in getting and using some of the video your members may } } have made on primitive life forms such as viruses, bacteria, etc. } } I would be also interested in } } acquiring from you a high quality beta-sp or preferable dv tape you might } } have of these studies as well as other studies you may have of microscopic } } life forms/elements dna, diatoms, etc. Full credit to your organization } } (and the videographer if desired) would be given in the final piece. I } } would pay for all shipping and handling costs. } } } } } } } } The video and images would be a small component of a much longer work. } } } } My last work recently screened at the Canadian Embassy in Washington, D.C. } } and I have had shows at the Museum of Modern Art in New York City and at } } many other prestigious venues around the world. I include my bio. below. } } } } Please get back to me regarding my request. } } } } } } Best Wishes, } } Oliver Hockenhull hereticfilms-at-home.com
} } BIO - 2001 Oliver Hockenhull } } } } Home Site: http://www.crosswinds.net/~ideoplastic/ } } (From this site link to various works in VRML, hypertext, etc.) } } } } Oliver Hockenhull is an award-winning media artist, director, communication } } theorist, and lecturer. } } } } He has completed five feature length works and numerous dramatic and } } experimental shorts. The films have shown internationally at such film } } festivals and venues as -(selected) The Kerala-India International Film } } Festival, The Montréal World Film Festival, The Leipzig International } } Documentary Film Festival, The Vancouver International Film Festival, The } } European Media Arts Festival, The Nouveau Cinema Festival , The } } International Documentary Film Festival of Amsterdam, The Museum of Modern } } Art in New York City, the National Gallery of Art in Washington, D.C., and } } LACE (Los Angeles Contemporary Exhibitions). } } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } Scientific Ditrector, ICBR EM CORE } University of Florida Ph. 352-392-1295 } PO Box 118525 Fax 352-846-0251 } Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl }
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues } } We have a student who has been looking at biofilms on artificial } membranes and in the process found a strange-looking inclusion. If } there's anyone out there who has the time to have a look at this (it } is displayed on http://www.ru.ac.za/emu/lechbug.htm) and give any } suggestions this will be much appreciated. } } Regards } } Rob } } } ======================================================= } Robin H Cross (Mr) } Director : Electron Microscopy Unit } Rhodes University, PO Box 94, Grahamstown, South Africa } tel: +27 46 603 8168/9, fax: +27 46 622 4377 } email: r.cross-at-ru.ac.za } http://www.ru.ac.za/emu/index.htm } } ** remember ICEM-15 in Durban in September 2002 ** }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
} Hello listers, } I recall seeing that thread late last week about the business opportunity. } This just came in from a Canadian news source, CBC. } Nick } RCMP make arrests in international phone scam } WebPosted Tue Jul 10 17:27:29 2001 } TORONTO - The RCMP, along with the FBI and the U.S. Secret Service, have } cracked a multi-million dollar scam originating in Nigeria and arrested } three people in the Toronto area. } More than 300 people around the world - most of them Americans - were } duped into giving money to a fake organization. The RCMP say some people } lost more than $50,000. Others lost millions. } Police say the fraud was sophisticated, high tech and organized. } Typically, victims received a letter either by mail or fax allegedly from } officials of a large Nigerian institution, such as the Nigerian National } Petroleum Corporation or the Central Bank of Nigeria. } The letters would contain an urgent request for help in transferring } millions of dollars out of Nigeria to financial institutions in the } victim's country. If the victims would help, they'd be in line for a } handsome profit, in some cases as much as 30 per cent of the monies being } transferred. } The letter says foreign accounts are needed and victims were asked to wire } personal banking information and $10,000 US to cover administration fees. } 'Boiler room' in Toronto } Then, victims were contacted by a person posing as a merchant banker of } the Central Bank of Nigeria. They were told the funds were now in a } clearing house or mercantile bank in North America. } Phone numbers given for the fake banks actually rang in a "boiler room" in } the Toronto area. } The scam continued with the victim asked to pay taxes, duties and other } fees, all with the promise of a windfall, until the victim was bankrupt. } The RCMP began a co-ordinated effort with U.S. police forces in 1998. } According to an RCMP news release, police detachments in the Toronto area } have received at least one complaint a week for the past eight years from } victims who lost money in the scam. } Staff Sergeant Darryl Ross says victims lost more than money. Some } suffered health problems, mental breakdowns, divorce and the break-up of } their families. } }
Can anyone recommend a good/cost-effective type of wall shielding to eliminate 60 and 180 cycle interferences coming into an SEM room from recently-installed large motors on the opposite side of the wall (that can't be moved now)?
Much obliged, Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
As chairperson of the Traveling Poster Exhibit I am looking for judges for the Poster Award Program. Anyone who will be attending the MSA meeting at Long Beach and would be interested in helping please contact me. I need five judges for each discipline, Biological and Material Sciences. Thank you, Caroline Miller
Dear Goeff, Paraffin embedding works best on tissues that have some internal support, such as a collagen matrix (e.g., skin, etc.). If you think about the matter, the treatment to which tissues are subjected is harsh. Kidney tends to dehydrate, clear and infiltrate more slowly. Further, any imperfection in those steps can lead to a hard, fractious tissue. I understand what you want to do, and, while I should not overextend, I feel if I'm going to help with one, I may take the liberty of helping with all.
First. Whole kidney is very hard to infiltrate with paraffin without either segmenting it or removing the capsule (which is not always easy). I always, at least, opened the capsule. This is probably necessary, because as dehydration, clearing and infiltration proceed the capsule shrinks with the organ. If the capsule shrinks more, which is my conclusion on the matter, the internal pressure might be sufficient to retard the processes. Second. If 'First' is true, then the heat will essentially dry the parenchyma of the organ and leave you with the impression that it is 'hard'. It is interesting, that such blocks, when faced into the tissue always respond positively to immersion to glycerinated water (10-20%). Of course, all this manipulation does not leave one in the mood to try to finish with something quantitative, so some means to avoid the problem must be devised.
Solutions. 1. I agree with the frozen section idea, if you can take serial sections, then you can dye/stain and count every section if you wish or some statistical sample designed for glomeruli. 2. I would try a milder dehydration routine and vacuum embedding/infiltrating, at the lowest temperature, for 48 hours. My experience has been with whole organs, and these can cause problems. A milder dehydration does not take more than 1.5 the time but softens the steps (e.g., instead of 30,70,95,100 EtOH, benzene you could use 20% steps in EtOH followed by 1:2,1:1,2:1 (Benzene:EtOH) and 1:1(Benzene:Paraffin) sub-steps (no longer than 1.25hr) and all but the benzene:paraffin steps at 4oC.
NOTE: Tt has been said that the most arduous transition is to 70%EtOH, thus, if you accept that, then one ought to be able to leave tissue for long periods at higher concentrations. I once experimented with the final dehydration step in 100EtOH (and I mean 100%(200 Proof)) by placing a piece of tissue in absolute EtOH in my -10oC freezer with a pint bottle of 200 Proof. I changed the fluid each day for a week and then finished the processing. I saw no difference between the tissue processed at RT and that dehydrated at -10oC, but if I had to try a whole kidney, I would remember that brief experiment in the execution of the art of tissue processing if I had infiltration problems.
Solution (My own approach). I would take control kidneys, since rat kidney is unipolar as I recall, and take transverse segments of known thickness (single-edge razor blade) through the fixed kidney (laying on its side or held 'up' with the 'ends of the bean' down) being careful to isolate the segments in order. I would then process each segment between leaves of fine mesh, using vacuum infiltration for the shortest time and at the lowest temperature. Following installation of each segment in a block, I would carefully orient each block to permit me the largest return of serial sections {10um. The ribbons of serial sections would be placed on poster board in order. Knowing the mean diameter of glomeruli, I would choose a sub-serial sample of sections from each segment and mount them on slides, in order. Meanwhile, my notebook would be filling with the numbers of the sections from the series of each segment (A4-20-1)=Slide Label = Segment A, Start w/section 4, every 20th section, slide 1), etc. Each slide with a mounted series would be marked with a diamond scribe as 1,2,... and then as above - record in notebook. I would then stain each slide with the PAS (periodic acid Schiff) reaction for carbohydrates (BUT in kidney, really does a great job on glomeruli, and one can use a green filter to count and photograph). A nuclear counterstain is optional depending on your purpose, BUT then, there are hundreds of other, adjacent sections waiting for such elaboration if you need them. If the PA(to generate aldehydes from vicinyl glycols) and the S(especially!) are fresh and tested (20ml of S with a drop of formaldehyde to prove the color change from none to intense magenta), the PAS is almost entirely predictable and effortless(?). I would then count the glomeruli in each section or in a statistically predetermined sub-sample, multiply, and get ready for the pathologic specimens. NOTE: Tissue dryness can occur even with the best laid plans and execution, so I have one more suggestion - which will, of course, eliminate the PAS as an appropriate stain. Consider using Polyethylene Glycol(PEG) as an embedment. For those who have learned the method, it is easy and productive, though I know of no reconstructive or statistical methods that have been based on sections of PEG-infiltrated material. NOTE: In any morphometric analysis, it is the sampling method that makes the entire process meaningful or not. Take for example the alternate experiment on a sheep. Would you have the same plan as you proposed to us? What if you wanted to compare mouse and elephant with respect to the structure of a single cell type. What are the statistical implications of the problem? [RHETORICAL QUESTION!!!!!!!!] What you might really like is to be able to take 10 random sections from each serially sectioned segment of a kidney and demonstrate not only the differences between normal and pathologic organs but also the presence or absence of any variation in the distribution of pathology in any organ. Would you expect the same result in a mouse or rabbit which also have unipolar kidneys? What about your expectation in a multipolar kidney like yours? How about a horseshoe multipolar like mine?
I have a protocol for PAS which you may have for the asking, but it is to be found in any version of Pearse, A.G.E. and many others for the looking. And by elaborating as I have done, I remain open to further questions that will be welcome.
Regards,
Fred Monson Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Geoff McAuliffe } Sent: Tuesday, July 10, 2001 10:47 AM } To: CachatF-at-aol.com } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: stereology on thick sections } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } "CachatF-at-aol.com"-at-sparc5.microscopy.com wrote: } } } I am interested in counting the total number of glomeruli in the kidneys } in } } } rats with obstructive uropathy. To perform stereology, I have to cut } thick } } } sections at 20 micrometers. I found that the kidney itself breaks down } and } } } get very damaged at this thickness (kidneys fixed in 10% formalin and } } } embedded in parrafin), probably because the kidney is much harder than } the } } } paraffin. In the litterature, some people embed kidneys in glycol } } } methacrylate. It might be better (harder), but then I do not know how } to } } cut } } } my sections which are about 2.5 to 3 cm long, if I need a } glass/diamond } } knife } } } that long. Any ideas about cutting thick sections in paraffin?, or is } it } } } worth re-embedding in glycolmethacrylate, and any ideas about cutting } my } } } serial sections at 20 micrometers in that resin? } } } } } } Thank you a lot for your help! } } } } } } F. Cachat } } } University of Virginia } } } fc6b-at-virginia.edu } } How about frozen sections? Fix in formalin, cryoprotect with ascending } concentrations of sucrose (10%, 20%, 30%) then snap freeze and section at } 20 } microns on a sliding microtome. Collect sections in ice-cube trays to keep } them } in order. } In my experience, thick paraffin sections are much more difficult than } thinner } ones. Embedding tissue in methacrylates (and other plastics) is usually } intended } for thinner sections (1-3 microns). } } Geoff } -- } ********************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane, Piscataway, NJ 08854 } voice: (732)-235-4583; fax: -4029 } mcauliff-at-umdnj.edu } ********************************************** } } } }
Research Associate position(s) available immediately to study roles of calcium dynamics during somite formation and myofibril assembly. Primary training in real-time digital fluorescence microscopy (both wide-field and confocal) using calcium tools, FP-tagged constructs, and FRET approaches. Experience in protein biochemistry or molecular biology highly desirable. Salaries commensurate with qualifications and experience, start date is negotiable. Research and position descriptions can be found at http://biology.uark.edu/ferrari. Research and resources at the University of Missouri - Kansas City School of Biological Sciences can be reviewed at http://sgi.bls.umkc.edu/research.html. Please send cover letter, CV, and names of three references to either address below or, preferably, attach via e-mail: ferrari-at-uark.edu. The University of Missouri-Kansas City is an affirmative action/equal opportunity employer.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Michael B. Ferrari Department of Biological Sciences University of Arkansas Fayetteville, Arkansas 72701 Ph: 501-575-6372 Fax: 501-575-5349 http://biology.uark.edu/ferrari
*****Note address change effective Jan. 2002***** School of Biological Sciences University of Missouri - Kansas City Kansas City, Missouri 64110-2499 Ph: 816-235-2247 Fax: 816-235-5595
} Peggy Bisher wrote: } } } One of the scientists here wants to give a demonstration involving } } vortices and fluids. He has a few short demonstrations to show the } } kids, but I told him I think he needs more to keep their interest and } } to keep them busy. He has five groups of 6 children for about an } } hour. } Hi, Peggy -- } The Exploratorium, an excellent hands-on science museum in San Francisco, sells manuals on building interactive demo equipment; do a web search. For example, here's a description of their optics manual (from the MICRO bibliography - URL below): ---------------------- Doherty, P. and Rathjen, D. 1995 The Magic Wand and other Bright Experiments on Light and Color 125pp, 7.5x9", $10.95. ISBN 0-471-11515-0 John Wiley & Sons, NY The Exploratorium in San Francisco is famous for its interactive science exhibits. They have published a series of "science snackbooks" that give adults detailed instructions on how to duplicate those exhibits for classrooms or science museums. This one collects 25 easy-to-construct optics demonstrations that investigate reflection, refraction, polarization, and magnification. Many could be good science fair projects. All grade levels. RECOMMENDED ----------------------
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Is anyone familiar with the Golgi-Cox Method for LM of brain? Any help would be appreciated! The method I was referred to uses Amfix(?) to decolorise the background. Has anyone heard of this and where can I order it? Or if anyone has a better method that has worked for them, please let me know.
Thanks! Peggy Sherwood -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
We've recent acquired a Bomar SPC-900/EX Critical Point Dryer, but of course it didn't come with an operator's manual. Does anyone out there have a manual for this instrument, and if so, would you be willing to send or fax me a copy? Is this company still in business?
Thanks in advance,
Steve
Steven Verhey, Ph.D Assistant Professor Department of Biology Central Washington University Ellensburg, WA 98926 509.963.3431 office 509.963.2730 fax verheys-at-cwu.edu
Unfortunately, good and cost effective may not be go together when speaking of large area magnetic shielding. The best material for magnetic shielding is known as Mu-Metal. This material is typically supplied fully annealed which provides the most effective shielding. FWIW, If strained by forming, it is common to re-anneal. This probably won't affect you if you simply "wallpaper" with sheet stock using contact cement. I would bond joints well (pop rivets, solder?). The web site for a supplier of Mu-Metal is: http://www.advancemag.com/
A poor second choice might be fully annealed silicon steel used in electrical transformers.
Regards, Woody White McDermott Technology, Inc. ---------------------------------------------------------------------
} -----Original Message----- } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] } Sent: Wednesday, July 11, 2001 11:07 AM } To: microscopy-at-sparc5.microscopy.com } Subject: magnetic field shielding } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi colleagues, } } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Much obliged, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) } } }
I remember receiving the same scam from the mail. I am surprised that people can and will succumb to these things. Oh well, stupid is as stupid does.
Earl ----- Original Message ----- } From: "Nicol Aitken" {nicol-at-semiconductor.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, July 11, 2001 7:05 AM
} } Peggy Bisher wrote: } } } } } One of the scientists here wants to give a demonstration involving } } } vortices and fluids. He has a few short demonstrations to show the } } } kids, but I told him I think he needs more to keep their interest and } } } to keep them busy. He has five groups of 6 children for about an } } } hour. } } } Hi, Peggy -- } } } The Exploratorium, an excellent hands-on science museum in San Francisco, } sells manuals on building interactive demo equipment; do a web search. For } example, here's a description of their optics manual (from the MICRO } bibliography - URL below):
I would like to second Caroline's recommendation for the exploratium. They have put a lot of the interactive demos on their website,check out: http://www.exploratorium.edu/snacks/snacksbysubject.html There are other great informal science education tools on their website, try using their search engine to find what you need.
Bomar is a name that will live in infamy.....at least in this facility. More than 15 yrs ago, the former director returned a CPD unit to Bomar for repair. After not hearing from them for many months, and after her phone calls were not returned, she demanded the return of the unit. No response. The company had gone bankrupt, its assets were seized and our unit was gone. We did file a lawsuit against the company but were told to forget pursuing the matter since it would cost more than the unit was worth. If you do find that they are in business again, let me know. I will look around to see if any information on the CPD.
Good luck......
JB
} We've recent acquired a Bomar SPC-900/EX Critical Point Dryer, but } of course it didn't come with an operator's manual. Does anyone out } there have a manual for this instrument, and if so, would you be } willing to send or fax me a copy? Is this company still in business? } } Thanks in advance, } } Steve } } Steven Verhey, Ph.D } Assistant Professor } Department of Biology } Central Washington University } Ellensburg, WA 98926 } 509.963.3431 office } 509.963.2730 fax } verheys-at-cwu.edu
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
We routinely cut 20-50 micron sections from methacrylate blocks for stereology and as far as we are concerned it is the only way to go for stereology studies. We use technovit 7100 kits and deal with samples as large as whole ovaries from sheep.If the whole ovary is to large we simply cut it in half and process and section seperately, for stereology studies we simply count the 2 halves seperately and the sum of the 2 halves = the whole organ.The cutting is a bit of an art but with practice becomes fairly easy, fortuneately we are blessed here with an artist and the following is a description of how she does the cutting.We have also used a number of different stains on the thick sections, if you decide to go down this path and need any further info don't hesitate to e-mail us, alternatively look for any references by HJG Gundersen, in my opinion the guru of stereology, a good review article is in APMIS 96:1988 p 379-394 and p857-881.
The following is how we cut thick sections (30 Microns), using Technovit Resin. Before cutting thick sections, blocks have to be completely hard, either been made for months or hardened in a 37 degree oven for 3-5 days. Unlike paraffin sections which have a right and wrong side, Resin/Plastic sections can be placed in the waterbath on either side, so for this reason I cut the right hand corner off the block, so I can regulate the mounting of sections onto slides. I use a Microm Rotary Microtome HM 350 with a Ralph Glass Knife with a concave knife profile, knives are made on a LKB Histo Knifemaker. The microtome is programmed with a cutting window (selected according to the size of the block) and a cutting speed (selected speed is 2.2). As I mainly do serial sections I have a continuous stroke programmed on the microtome (this means that I pick up a section every 5-10seconds), the microtome is controlled with the foot/hand switch. My waterbath is a large glass staining dish with distilled water at room temperature which is placed in a black box (so that you can see the sections on the bottom of the waterbath). I also have a small glass beaker with distilled water and a pipette, this is so that I can put water onto the glass knife as the section is being cut. Cut sections are taken to forceps and placed into the waterbath were they rest on the bottom of the waterbath until they are picked up onto cleaned glass slides and placed into a 60 degree oven for at least 30 minutes (sections are very wrinkled at first but straighten out with the heat from the oven).
Hope this is of some help
Peter Smith Reproduction Group AgResearch Wallaceville Research Centre P.O. Box 40063, Upper Hutt, New Zealand
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I am not familiar with the Golgi-Cox method, but if it involves silver in any shape or form, then the "Amfix" most probably refers to a photographic rapid fixer (ammonium thiosulphate). As far as I know this is still available, but Agfa Agefix and Ilford Hypam would be equivalent. Kodak Unifix would probably work as well, but it does contain a photographic emulsion hardener.
Jan Coetzee
Peggy Sherwood wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all! } } Is anyone familiar with the Golgi-Cox Method for LM of brain? Any } help would be appreciated! The method I was referred to uses } Amfix(?) to decolorise the background. Has anyone heard of this and } where can I order it? Or if anyone has a better method that has } worked for them, please let me know. } } Thanks! } Peggy Sherwood } -- } Peggy Sherwood } Lab Associate, Photopathology } Wellman Laboratories of Photomedicine (W224) } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-724-4839 (voice mail) } 617-726-6983 (lab) } 617-726-3192 (fax) } sherwood-at-helix.mgh.harvard.edu
-- Prof Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Dee Breger wrote: Can anyone recommend a good/cost-effective type of wall shielding to eliminate 60 and 180 cycle interferences coming into an SEM room from recently-installed large motors on the opposite side of the wall (that can't be moved now)?
Hi, Have you as an alternative considered to use the magnetic field cancellation systems that are available nowadays? The first I found by searching the web for "magnetic field cancellation" was:
http://www.spicerconsulting.com/sc12.htm
but there are quite a few other producers.
Best regards, Jorgen.
{:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::}
Joergen B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I am too confront with this problem. But we can move the SEM (witch we have just moved... in a electromagnetic "dirty" room!).
As Woody said it, mu-metal is the best way, but with terrible difficulties and cost. One of our collegues, magnetism specialist, proposed to use thick aluminium or cooper plates (10 mm or more thick). Aluminium (dural) has the big advantage to be light, copper is better for electrical continuity, but heavier and expensive (with 10 to 15 mm thick...), soft iron or silicon iron is not too expensive, but heavy. You must build a box around the SEM, with a door on entry lock side and a good electrical continuity. It looks nice ! It's not a simple problem, but as consolation, remember that you can recycle the plates when the interferences are eliminated. But of course the best way is to eliminate the source of interferences.
By the way, did you mesure the AC field ? In case, how much do you have ? I am interested on the solution you will find. Our building is VERY noisy.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Unfortunately, good and cost effective may not be go together when speaking of large area magnetic shielding. The best material for magnetic shielding is known as Mu-Metal. This material is typically supplied fully annealed which provides the most effective shielding. FWIW, If strained by forming, it is common to re-anneal. This probably won't affect you if you simply "wallpaper" with sheet stock using contact cement. I would bond joints well (pop rivets, solder?). The web site for a supplier of Mu-Metal is: http://www.advancemag.com/
A poor second choice might be fully annealed silicon steel used in electrical transformers.
Regards, Woody White McDermott Technology, Inc. ---------------------------------------------------------------------
} -----Original Message----- } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] } Sent: Wednesday, July 11, 2001 11:07 AM } To: microscopy-at-sparc5.microscopy.com } Subject: magnetic field shielding } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi colleagues, } } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Much obliged, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) } } }
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 10 - October 19, 2001
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2150 (Includes room and board, text, handouts, supplies)
Application Deadline: July 25, 2001
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual
At 3:53 PM -0400 7/11/01, Peggy Sherwood wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all! } } Is anyone familiar with the Golgi-Cox Method for LM of brain? Any } help would be appreciated! The method I was referred to uses } Amfix(?) to decolorise the background. Has anyone heard of this and } where can I order it? Or if anyone has a better method that has } worked for them, please let me know. } } Thanks! } Peggy Sherwood } -- *****************
Hi Peggy, Here is the Golgi-Cox protocol, with formulae. Its out of:"Histological & Histochemical Methods: Theory & Practice" by JA Kiernan (1981)Pergamon Press.
A. Golgi-Cox Fixative-prepared from 3 stable stock solutions: (i) 5% aqueous mercuric chloride (HgCl2) (ii) 5% aqueous potassium dichromate (K2Cr2O7) (iii)5% aqueous potassium chromate (K2CrO4)
Working s olution} Mix, IN ORDER (i) 20ml (ii) 20ML Water: 40ml (iii) 16ml Mix before useing.
B. Alkaline developer Strong ammonia solution: (17% NH3) 5 ml Water 95 ML Mix just before using.
Procedure:
1. fix pieces of nervous tissue (not more than 10mm thick) in the working sol'n of the golgi-Cox fix for 6-8 weeks in a tightly capped container at 37 deg. C. Pour off fix and replace with fresh after first day. 2. Wash in water, many changes, for 6-8 hours, then dehydrate and embed in nitrocellulos. Cut sections at 100 microns thick. 3. treat sections with the alkaline developer (sol'n B) for 2-3 minutes. 4. Wash in water, dehydrate, clear & mount in a resinous medium. (see NOTE)
Result: some neurons & neuroglial cells, including cytoplasmic processes--black. Background, pale yellow.
NOTE: A mixture of 33ml chloroform, 33ml xylene and 33ml abs,. alcohol is preferred as the lat stage of dehydration. This can be followed by clearing for 10-15 min in creosote or in cedarwood oil, followed by a rinse in xylene.
thae black color fades with time if a coverslip is applied, so it is usual to mount sections in THICK canada Balsam, and allow to set at 40-45 deg C without a coverslip. Such preps. are stable for many years. If a synthetic mounting medium is used (such as DPX), coverslips may be applied, but fading occurs after about 6 months.
Its a lot of work, using reagents most of us are not familiar with, and which will make your Health & Safety officers cringe! good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
We are looking for a Philips 400 series hot stage. The intent is to make some modifications to it so that even one that is broken (depending upon what is broken) might be acceptable. If you have one that you might be willing to part with, please communicate to me its state and under what conditions you might let it go.
Richard
Richard Vanfleet Assistant Professor Advanced Materials Processing and Analysis Center (AMPAC) and Department of Physics
Mail: AMPAC University of Central Florida 12443 Research Parkway, Suite 404 Orlando, FL 32826
I thought this might be worth drawing to your attention if you scan e.m. negatives in with a flat bed scanner.
We currently use an Epson Perfection 1200 with transparency adapter. I know the spec is considered to be pretty low in terms of O.D., greyscale and DPI but it produces very good pictures about 98% of the time. One problem that I have encountered, however, from time to time is one or rarely more than one extremely fine dark lines appears on the scanned image. This can be very annoying because they are only visible if you look very carefully at an enlarged image. I had assumed that there was a problem with one or two points on the CCD scanner detector array because the lines were straight and at right angles to the scanner head.
The lines are actually caused because this particular scanner has a small reference slot to calibrate the CCDs for fluctuations in the light. If a small particle of dust falls in this area of the glass top it causes the the scanner to overcompensate for the CCD on the corresponding point of the scanner head. This usually results in a very fine white line on the negative image or black on the positive image. I confirmed this using a series of varying density dots on film and got a perfect series of lines.
I'm sorry if people knew this already but thought it just might save someone tearing their hair out or throwing a perfectly functional scanner away. I don't know if some of the other flatbed glass topped scanners with transmitted light sources use a similar calibration system but it's worth taking extra care if they do.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland SUNDERLAND UK
One other very interesting aspect of the scam. The letters sent via snailmail used counterfeit (forged) stamps. So many of these letters were sent that copies of these fakes still show up in packets of stamps sold to collectors. And the envelopes with stamps on them (called covers in the hobby) are now considered to be relatively valuable collectors' items. (Oh, you mean it's obvious that I'm one of those stamp collectors? Too true.)
Roger On Wed, 11 Jul 2001 15:38:24 -0700, Earl Weltmer wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | I remember receiving the same scam from the mail. | I am surprised that people can and will succumb to these things. | Oh well, stupid is as stupid does. | | Earl | ----- Original Message ----- | } From: "Nicol Aitken" {nicol-at-semiconductor.com} | To: {Microscopy-at-sparc5.microscopy.com} | Sent: Wednesday, July 11, 2001 7:05 AM | Subject: RE: more on that scam | | | } ------------------------------------------------------------------------ | } The Microscopy ListServer -- Sponsor: The Microscopy Society of America | } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | } -----------------------------------------------------------------------. | } | } | } | } | } | } } Hello listers, | } } I recall seeing that thread late last week about the business | opportunity. | } } This just came in from a Canadian news source, CBC. | } } Nick | } } RCMP make arrests in international phone scam | } } WebPosted Tue Jul 10 17:27:29 2001 | } } TORONTO - The RCMP, along with the FBI and the U.S. Secret Service, have | } } cracked a multi-million dollar scam originating in Nigeria and arrested | } } three people in the Toronto area. | } } More than 300 people around the world - most of them Americans - were | } } duped into giving money to a fake organization. The RCMP say some people | } } lost more than $50,000. Others lost millions. | } } Police say the fraud was sophisticated, high tech and organized. | } } Typically, victims received a letter either by mail or fax allegedly | from | } } officials of a large Nigerian institution, such as the Nigerian National | } } Petroleum Corporation or the Central Bank of Nigeria. | } } The letters would contain an urgent request for help in transferring | } } millions of dollars out of Nigeria to financial institutions in the | } } victim's country. If the victims would help, they'd be in line for a | } } handsome profit, in some cases as much as 30 per cent of the monies | being | } } transferred. | } } The letter says foreign accounts are needed and victims were asked to | wire | } } personal banking information and $10,000 US to cover administration | fees. | } } 'Boiler room' in Toronto | } } Then, victims were contacted by a person posing as a merchant banker of | } } the Central Bank of Nigeria. They were told the funds were now in a | } } clearing house or mercantile bank in North America. | } } Phone numbers given for the fake banks actually rang in a "boiler room" | in | } } the Toronto area. | } } The scam continued with the victim asked to pay taxes, duties and other | } } fees, all with the promise of a windfall, until the victim was bankrupt. | } } The RCMP began a co-ordinated effort with U.S. police forces in 1998. | } } According to an RCMP news release, police detachments in the Toronto | area | } } have received at least one complaint a week for the past eight years | from | } } victims who lost money in the scam. | } } Staff Sergeant Darryl Ross says victims lost more than money. Some | } } suffered health problems, mental breakdowns, divorce and the break-up of | } } their families. | } } | } } | } | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
} } Is anyone familiar with the Golgi-Cox Method for LM of brain? Any } } help would be appreciated! The method I was referred to uses } } Amfix(?) to decolorise the background. Has anyone heard of this and } } where can I order it? Or if anyone has a better method that has } } worked for them, please let me know.
I am not familiar with any need to decolorize the background in most of the Golgi impregnation methods, the Cox variant included. OTOH, my experience has not been with wax embedding but only with the harder (celloidin, araldite, etc.) media do not require any clearing.
There is, however, a need to add an oxident, like NaOH, to the alkalinization step after impregnation and before embedding to prevent background blackening.
I have a Sorvall JB4 microtome that is not advancing at all. If anyone has any suggestions, please contact me. Thanks.
Phoebe
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Department of Physiological Sciences 264 McElroy Hall Oklahoma State University Stillwater, OK 74078 405-744-6765
My two cent's worth: as a former Spicer Consulting SC12 user, I was highly satisfied with the unit. We too had EMI from large power cables for water pumps and some other unknown source affecting our SEM images and the SC12 cleared it up. I don't remember how much the unit cost, but remember that is was less than a room full of Mu-Metal. With the added advantages of easy installation and if the SEM ever changes locations, the EMI shielding (the unit) can go with it.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dee Breger wrote: } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Hi, } Have you as an alternative considered to use the magnetic field cancellation } systems that are available nowadays? The first I found by searching the web } for "magnetic field cancellation" was: } } http://www.spicerconsulting.com/sc12.htm } } but there are quite a few other producers. } } Best regards, } Jorgen. } } {:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::} } } Joergen B. Bilde-Soerensen } Senior Research Scientist, Ph. D. } Materials Research Department } Risoe National Laboratory } DK-4000 Roskilde } Denmark } } e-mail: j.bilde-at-risoe.dk } phone: +45 4677 5802 (direct) } phone: +45 4677 4677 (switchboard) } fax: +45 4677 5758 } website: http://www.risoe.dk/afm/Personal/jqbi/jqbi.htm
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I am intrigued by the recommendation of the referenced magnetism specialist to use aluminum and/or copper plate. The principle in using high conductivity non ferromagnetic shielding (such as Al and Cu) is that of eddy current shielding. This is very effective for high frequency magnetic fields, but becomes worthless as you approach DC (except for the special case of super conducting material, which does shield DC fields). Line-frequency magnetic fields (50-60Hz) are typically dealt with via ferromagnetic shielding because of the relatively low frequency.
Presumably your specialist has done the calculations and feels that eddy current shielding will do the job. This suggests one of three things to me: (1) he knows the problems are due to relatively high frequencies; (2) your fields are very severe and he is only trying to knock them down to a more tractable level; or (3) he knows something I don't. I would personally be interested in learning more about how he arrived at this solution. Specifically, what level of attenuation does he anticipate achieving for 50 Hz magnetic fields with this kind of shielding?
Fred Schamber
Faerber Jacques wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am too confront with this problem. But we can move the SEM (witch we } have just moved... in a electromagnetic "dirty" room!). } } As Woody said it, mu-metal is the best way, but with terrible difficulties } and cost. One of our collegues, magnetism specialist, proposed to use } thick aluminium or cooper plates (10 mm or more thick). Aluminium (dural) } has the big advantage to be light, copper is better for electrical } continuity, but heavier and expensive (with 10 to 15 mm thick...), soft } iron or silicon iron is not too expensive, but heavy. You must build a box } around the SEM, with a door on entry lock side and a good electrical } continuity. It looks nice ! It's not a simple problem, but as } consolation, remember that you can recycle the plates when the } interferences are eliminated. But of course the best way is to eliminate } the source of interferences. } } By the way, did you mesure the AC field ? In case, how much do you have ? } I am interested on the solution you will find. Our building is VERY noisy. } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess } 67037 Strasbourg CEDEX } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } ---------- Forwarded message ---------- } Date: Wed, 11 Jul 2001 16:49:16 -0500 } } From: "White, Woody N." {nwwhite-at-mcdermott.com} } To: 'Dee Breger' {micro-at-ldeo.columbia.edu} , microscopy-at-sparc5.microscopy.com } Subject: RE: magnetic field shielding } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Dee, } } Unfortunately, good and cost effective may not be go together when speaking } of large area magnetic shielding. The best material for magnetic shielding } is known as Mu-Metal. This material is typically supplied fully annealed } which provides the most effective shielding. FWIW, If strained by forming, } it is common to re-anneal. This probably won't affect you if you simply } "wallpaper" with sheet stock using contact cement. I would bond joints well } (pop rivets, solder?). } The web site for a supplier of Mu-Metal is: http://www.advancemag.com/ } } A poor second choice might be fully annealed silicon steel used in } electrical transformers. } } Regards, } Woody White } McDermott Technology, Inc. } --------------------------------------------------------------------- } } } -----Original Message----- } } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] } } Sent: Wednesday, July 11, 2001 11:07 AM } } To: microscopy-at-sparc5.microscopy.com } } Subject: magnetic field shielding } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } } ---------. } } } } } } Hi colleagues, } } } } Can anyone recommend a good/cost-effective type of wall shielding to } } eliminate 60 and 180 cycle interferences coming into an SEM room from } } recently-installed large motors on the opposite side of the wall (that } } can't be moved now)? } } } } Much obliged, } } Dee } } } } } } } } } } *************************************************************** } } Dee Breger } } Mgr. SEM/EDX Facility } } Lamont-Doherty Earth Observatory } } 61 Route 9W } } Palisades, NY 10964 USA } } T: 845/365-8640 } } F: 845/365-8155 } } } } http://www.ldeo.columbia.edu/micro } } http://www.discovery.com/area/science/micro/micro1.html } } http://www.lsc.org/antarctica/front.html } } Journeys in Microspace (Columbia University Press, 1995) } } } } } }
Our supplier of negative films (Agfa) for the Philips CM-200 EM in the Microscopy Service of the University of Seville does not fabricate these negatives any more. According to Philips Spain, Kodak does not sell those negatives in Spain (Kodak Electron Microscopy 4489, Dimensions: 6.5 x 9 cm) and they do not give us an immediate solution (?).
Could somebody help us with this silly problem and let us know of a supplier close to Spain?
May be I am wrong, but aluminum is "transparent" to the magnetic fields (as well as copper). Since the physics course in the middle school, I had strong feelings that soft iron is a good material for shielding. In Russia we had special requirements for EM rooms (like "floating" basement and proper magnetic/electric shielding). As I remember, our EM room has iron mesh immersed into the walls/ceilings/floors. That mesh was properly grounded (copper sheet 1x1 m buried 5 meters deep into the ground, 20 meters away of the building with very thick copper "conductor"; this "conductor" was mounted across the perimeter of the room and was connected to the mesh at many points). It also seems to me that the thickness of shielding material do not so important. I think, described "shielding" schedule was designed mostly against electric fields, but it should works against magnetic fields as well. I would be happy to listen the comments of the specialist on this matter.
Sergey.
At 10:29 AM 7/12/01 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Does anyone know who makes them and/or sells them on a regular basis?
rtch
} } } } Any opinions of this $195 device? } http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=1164152801 } } - John } P.S. I have no association with this auction. } }
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Its the Kodak dealer that has chosen not to stock that size. We no longer stock boxes of 100 sheets because there was too little demand and its very expensive when film goes out of date. ProSciTech and half a dozen other EM suppliers would be delighted to supply you. Just ask for a quote including freight. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, July 13, 2001 6:30 AM, Julian Martinez-Fernandez [SMTP:Julian.Martinez-Fernandez-at-grc.nasa.gov] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleges: } } Our supplier of negative films (Agfa) for the Philips CM-200 EM in } the Microscopy Service of the University of Seville does not fabricate } these negatives any more. According to Philips Spain, Kodak does not sell } those negatives in Spain (Kodak Electron Microscopy 4489, Dimensions: 6.5 x } 9 cm) and they do not give us an immediate solution (?). } } Could somebody help us with this silly problem and let us } know of a supplier close to Spain? } } Many thanks, } Julian
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I need information about the deep focus of objetives:
Plan Neofluar, 2.5/0.075, inf/0.17
Ph1 Plan Neofluar, 10x/0.30,inf/0.17
Ph2 Plan neofluar, 20x/0.50,inf/0.17
Ph2 plan neofluar, 40x/0.75, inf/0.17
thank you for your help
___________________________________
Dr. José Luis Pech Pacheco PhD. Marine Ecology Image & Vision D. Instituto de Óptica, CSIC. Madrid, Spain Fax 0034915645557 _____________________________________
} Thanks to all who replied to my question about the DTSA software package. I } have downloaded it onto a Mac here. As yet, I don't have it fully working } as it doesn't want to import the EMSA files exported by our Noran Voyager } system. I'm still trying to working out what the problem is, but in } principle it seems to be working (except with my files).
FYI, there are two DTSA plug-ins for EMSA import. A version 1.0 and version 1.1. Check to make sure your are using the correct one. If you still have problems, You can also e-mail me some of your Voyager EMSA files and I can tell you what is the problem. I'm well versed with the internals of DTSA and that of EMSA file formats.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Group, I just received a product release on a new product, one that I had not heard of before, that may be of interest to a number of you. The product, called ScanScope T50, from Aperio Technologies converts all information from a microscope slide into a common digital format. They say, for example, that a typical tissue section can be digitized at 54,000 pixels per inch. Their web site is www.aperiotech.com Obvioulsy I (unfortunately)have no financial or other interests in this company or its products. Don Grimes, Microscopy Today
I had seen this item a while back and may have said something to the listserver. At the time, I thought that it would have worked nicely on the eyepiece on a microprobe and perhaps on the binoculars of a TEM. I had gotten some information on it and here is the contact information:
International Micro-Vision Inc. 667 El Camino Real Redwood City, CA 94063-1317
I went to the web site. Not very impressive. Their big product digitizes all the info on a slide without a conventional microscope. OK, nice idea but don't you think they could have posted at least one image! No data of any type at the site.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have not solved our problems completely but will share some things I have "heard". If you decide to go with the mu-metal, it should be easier and cheaper to build a small enclosure around the actual source (motors) than to shield the EM or entire room. This assumes you are confident that the new motors are the source; i.e., the SEM was fine before their installation. Anyway, this is how I was advised to proceed; unfortunately we could not pinpoint a single source, so I haven't actually tried it. Often, the advice you may recieve from "magnetism experts" and shielded-room experts is better suited to the Earth's steady field than it is to A/C fields.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Wednesday, July 11, 2001 11:07 AM, Dee Breger [SMTP:micro-at-ldeo.columbia.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi colleagues, } } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Much obliged, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) }
In a message dated 7/11/2001 3:14:47 PM Mountain Daylight Time, micro-at-ldeo.columbia.edu writes:
} Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)?
Can you be more specific on what you mean by cost-effective by providing an actual dollar amount that you could afford? Otherwise, many of the suggestions being made may be completely impractical based on funding concerns.
The reason I suggest this is that the last I heard, the commercial magnetic field canceling systems cost from ~$10k to ~$25k. Also, one person has said that covering a room in mu-metal may be of a similar cost.
A less expensive mu-metal solution may be to build a box around the column, or even just around the chamber. Years ago, JEOL sold such a mu-metal box that surrounded just the chamber of a 840A SEM (on 4 full sides plus the top and bottom where it stopped at the perimeter of the column and chamber, respectively). At the time, it cost about $6k from JEOL. Using the JEOL mu-metal box (which was about 2mm thick, if I recall correctly), reduced the interference on the beam by about a factor of 3. (We were doing lithography at the time, so it was easy to compare the amplitude of the interference in the patterns that were written.)
I know one person who designed a similar box at a substantially lower cost using an in-house machine shop. The key in that case is to properly anneal the mu-metal after it has been fabricated.
Regarding the idea of using Cu or Al, I agree with the previous postings that question its usefulness for line frequency magnetic fields, although I haven't done any calculations to evaluate the suggestion.
I have also heard of soft iron being used, which should be relatively inexpensive, but will be very heavy because of the increased thickness that would be required to match the shielding of mu-metal.
Could you build a heavy, ugly, and cheap box around the motors???
The lowest cost solution may be to move the SEM, if you have a suitable room available. A professional move may cost thousands, but doing it yourself is a reasonable option, if you know about shipping bolts and scintillators.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Try mandel Scientific , Canada if you cannot find it closer. e-mail info-at-mandel.ca
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Julian Martinez-Fernandez {Julian.Martinez-Fernandez-at-grc.nasa.gov} 07/12 4:30 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleges:
Our supplier of negative films (Agfa) for the Philips CM-200 EM in the Microscopy Service of the University of Seville does not fabricate these negatives any more. According to Philips Spain, Kodak does not sell those negatives in Spain (Kodak Electron Microscopy 4489, Dimensions: 6.5 x 9 cm) and they do not give us an immediate solution (?).
Could somebody help us with this silly problem and let us know of a supplier close to Spain?
In a message dated 7/13/01 10:55:26 AM, PhillipsT-at-missouri.edu writes:
} I went to the web site. Not very impressive. Their big product } digitizes all the info on a slide without a conventional microscope. } OK, nice idea but don't you think they could have posted at least one } image! No data of any type at the site.
I agree that there is no info whatever on the site. Also, note that 54000 points per inch is 1/2 micron. Certainly that is not real resolution, and one wonders how with anything approaching that optical resolution they can assure good focus. It takes some serious mechanical engineering and software to scan a whole slide to collect and tile images. I've seen it done, using good microscopes and stages with well designed optics and a digital camera. I'd have to see some real examples before believing anything from this outfit.
I am a newcomer to this field, trying to differentiate between autofluorescent lignin, callose, cutin and suberin in LR-white embedded plant material. My question is: Is there a way of identifying one of these substances by selectively quenching the autofluoescence of the others ? Can the colour of the emission be used as a tool for differentiation ? I tried Berberinsulfat but it stains lignin and suberin. Neutralred stains the hydrophobic domain of suberin but is also used as a fluorochrome for lipids in general and therefore probably stains cutin as well.
Below is the result of your feedback form. It was submitted by (tcote-at-medicacorp.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, July 13, 2001 at 12:35:36 ---------------------------------------------------------------------------
Email: tcote-at-medicacorp.com Name: Tyler Cote
Organization: Medica Corporation
Education: Undergraduate College
Location: Bedford, MA 01730
Question: Is is it possible to get clear images of white and red blood cells using an air objective ?
I am curently working on a hematalogical assistant that uses a digital camera to image white and red cells. The red cells seem to be acting like little lenses. Most red cells appear to have a small dark band around them. Upon further investigation it appears this band is actually a series of colored bands.
When I wet the slide with a liquid that has an index of refraction closer to that of the cells, the bands disapear. I am still using an air objective, just looking at a wetted slide.
Is this common ?
Is there a way to get around having to wet the slides ?
Any help you can give is greatly appreciated. Thank you.
Below is the result of your feedback form. It was submitted by (joelgerardo-at-correoweb.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 11, 2001 at 16:22:19 ---------------------------------------------------------------------------
Email: joelgerardo-at-correoweb.com Name: joel gerardo diaz M.D.
Organization: medicine faculty unam
Education: Graduate College
Location: mexico city , fedeeral district City, State, Country
Question: I need a manual of Quality Control in the manufacturation of Colposcopes and Microscopes ,I will be very grateful for your attention, sincerely. Joel gerardo Diaz Sanchez M.D.
First, I must precise again that I am not a specialist of magnetisme ! I am only confronted with the same problem of electromagnetic interference. So I asked this collegue which has answered and calculated only with the hands.
What he said, and others have said it too, is than mu-metal is the best, soft iron could work too. About Al or Cu, it's of course only efficient with AC field. The goal is to disipate the AC electromagnetic filed in a good conductor by Foucauld current. 50 or 150 Htz is not DC, even it is not "high frequency". As a test is sometimes as good as much calculations, we have tested with a 10 mm thick Al plate, 500 x 500 mm large, aproching the SEM, and we have seen the distorsion reducing, but not been supressed. I don't know if an Al box would be sufficient to put down enought the interfernce. As we can move the SEM in an other room, we didn't try further. The advice of my collegue was : "If you can't supress the source, try to move the SEM ; shielding is often risky and expensive". That what other have soon said on the list.
So about your suppositions (mail from F. Schamber):
} } Presumably your specialist has done the calculations and feels that eddy } } current shielding will do the job. This suggests one of three things to } } me: (1) he knows the problems are due to relatively high frequencies; } } (2) your fields are very severe and he is only trying to knock them down } } to a more tractable level; or (3) he knows something I don't. I would } } personally be interested in learning more about how he arrived at this } } solution. Specifically, what level of attenuation does he anticipate } } achieving for 50 Hz magnetic fields with this kind of shielding?
it is the second witch is right ! We had 20-25 mGauss, 50-150 htz at the sample level . That is much more that the 3 mG max recommended.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
In a message dated 7/11/2001 3:14:47 PM Mountain Daylight Time, micro-at-ldeo.columbia.edu writes:
} Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)?
In my previous reply, I forgot to mention that a listing of suppliers for vibration and magnetic field canceling systems as well as suppliers for mu-metal enclosures can be found at "www.jcnabity.com/links.htm".
(If anyone knows of other companies that should be listed, please let me know.)
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Ian; I had the same import problem with Noran files. It seemed that the Noran interpretation of the EMSA file spec is different than the interpretation by the authors of DTSA. --John Mardinly
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] Sent: Friday, July 13, 2001 4:54 AM To: Microscopy Discussion Group
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I am currently evaluating EDS packages for my Philips XL-30 ESEM with LaB6 and wanted to poll the list members for their opinions, be they positive or negative. How do members deal with the beam spread issues? Can we trust the results? What about water vapor and ice build up? As a multi-user facility how easy is it to get the data out to the researcher these days? What about service quality?
I will post a summary so if you prefer your responses excluded, please let me know.
Thanks
Scott Whittaker SEM Lab Manager National Museum of Natural History Smithsonian Institution 10th & Constitution Ave, NW Washington DC 20560-0104 202-357-1651
The NORAN EMSA files follow the specifications fully and support all varieties of EMSA options available. The problem is that DTSA only reads one type of EMSA file format (X,Y pairs of data) which is not the default condition of the VOYAGER files. The default format for VOYAGER stores only the channel data in 5 columns. The default file format for VANTAGE was changed to be X,Y pairs which is consistant with DTSA.
There is a solution for VOYAGER. You can save the EMSA format compatible with DTSA by using the shell command: emsa_io -x=1 -w={path to store to} -s={spectrum name to store} example: emsa_io -x=1 -w=$HOME/spectra/myspec.emsa -s=spectrum1
When the next version of VOYAGER software is released the default condition for the emsa_io program will be converted to the same as VANTAGE which will make VOYAGER's default version of EMSA file format consistant with DTSA.
Greg Fritz
} -----Original Message----- } } From: Mardinly, John [mailto:john.mardinly-at-intel.com] } Sent: Monday, July 16, 2001 11:08 AM } To: 'Ian MacLaren'; Microscopy Discussion Group } Subject: RE: DTSA } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ian; } I had the same import problem with Noran files. It seemed that the } Noran interpretation of the EMSA file spec is different than the } interpretation by the authors of DTSA. } --John Mardinly } } -----Original Message----- } } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] } Sent: Friday, July 13, 2001 4:54 AM } To: Microscopy Discussion Group } Subject: DTSA } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks to all who replied to my question about the DTSA software package. I } have downloaded it onto a Mac here. As yet, I don't have it fully working } as it doesn't want to import the EMSA files exported by our Noran Voyager } system. I'm still trying to working out what the problem is, but in } principle it seems to be working (except with my files). } } As helpfully pointed out by Scott Wight at NIST, the latest version resides } on their server at } http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm } } Thanks again } } Ian MacLaren } Max Planck Institut f¸r Metallforschung, Seestraþe 92, } 70174 Stuttgart, Germany } Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk } Home page: http://members.tripod.co.uk/IanMacLaren/ } } }
I am not sure if anyone mentioned this, but there is a difference in using DTSA and the EMSA file handler plug-ins versions 1 and 2. Two did not work for me at all in any of the EMSA formatted files I tried, but version 1 did. I was using the power version of DTSA.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, July 16, 2001 12:08 PM To: 'Ian MacLaren'; Microscopy Discussion Group
Ian; I had the same import problem with Noran files. It seemed that the Noran interpretation of the EMSA file spec is different than the interpretation by the authors of DTSA. --John Mardinly
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] Sent: Friday, July 13, 2001 4:54 AM To: Microscopy Discussion Group
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I am not sure if anyone mentioned this, but there is a difference in using DTSA and the EMSA file handler plug-ins versions 1 and 2. Two did not work for me at all in any of the EMSA formatted files I tried, but version 1 did. I was using the power version of DTSA.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, July 16, 2001 12:08 PM To: 'Ian MacLaren'; Microscopy Discussion Group
Ian; I had the same import problem with Noran files. It seemed that the Noran interpretation of the EMSA file spec is different than the interpretation by the authors of DTSA. --John Mardinly
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] Sent: Friday, July 13, 2001 4:54 AM To: Microscopy Discussion Group
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
} To All following this lead, Is anyone concerned about the probability that the treatment of women described by several of the listers is illegal? Treating a woman like a criminal or a delinquent child who needs to be closely monitored as to her "motherhood" status is not only ridiculous, but descriminatory and hopefully illegal. It is the responsibility of each individual in a laboratory to watch out for his or her own safety and therefore the business and responsibility of each woman in the lab to watch out for the safety of her unborn child. We are all well past the kindergarten stage and should no longer require a hand to hold while crossing the street. Incensed in Iowa, Kristen
} to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified soldering } iron) has not worked well so far, any ideas how to eliminate xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
The Associate Director position at the University of Missouri Molecular Cytology Core (LM imaging facility) remains open.
This position involves training clients, developing new methodologies, maintaining instrumentation and managing junior staff members. Although many applicants will have a PhD in life sciences, we would be happy to consider individuals without a PhD but who have extensive experience in the relevant techniques. Opportunities for individuals with PhD's to conduct independent or collaborative research can be negotiated but the position is not tenure-track. The salary range for this position is $30,257 -$54,663.00 and is 100% benefit eligible. Opportunities for professional development will be encouraged.
Our facility includes a BioRad Radiance 2000 Confocal Laser Scanning Microscope system equipped with a Krypton/Argon laser (488 nm & 568 nm excitation lines) and Red laser diode (638 nm) laser mounted for highest sensitivity in the Keller configuration (bottom port) of an IX 70 Olympus inverted microscope. Simultaneous acquisition of three different fluorescent labels, as well as a high quality Nomarski image, is possible. Appropriate objectives, including water immersion and long working distance objectives, are available. An adjacent PC is equipped with the popular image analysis software package MetaMorph (Universal Imaging). For wide-field fluorescence work, we have another Olympus IX-70 inverted microscope equipped with a Photometrics Sensys cooled CCD camera, a Ludl filter wheel on the excitation side and a Ludl z axis stage motor, and a host of Chroma HQ single, double and triple fluorochrome filters. The Windows-based PC that controls this system includes the Vaytek MicroTome deconvolution software package that does Nearest Neighbor, No Neighbor, Constrained Iterative and Measure PSF deconvolution algorithms. It runs under Image Pro Plus (Media Cybernetics) for the image acquisition and 2D image analysis. For 3D work, we use MetaMorph. This system is also equipped with a CRI color filter to allow the acquisition of high quality color bright field images. Objectives and condenser apertures for high resolution phase contrast and differential interference contrast (DIC, Nomarski) are available. We also have a Nikon Optiphot upright photomicroscope with bright field, phase, DIC and fluorescence optics. This system is coupled to a Spot2 chilled CCD digital camera and a second copy of the MetaMorph image analysis and processing software. The main laboratory of the MCC includes a Reichert Frigocut 2800 cryostat for generating 2-100 µm thick frozen sections and a Reichert UltraCut S ultramicrotome for preparation of semi-thin (0.5 -1 µm) LM sections and ultra-thin EM sections. The MCC also contains two Apple workstations with extensive software for image manipulation and presentation graphics. For publication quality output, the Core is equipped with a Polaroid ProPallette 7000 film recorder and a Fuji Pictrography 3000 for photographic quality color prints. In the last year, our facility attracted researchers from the laboratories of over 100 different principal investigators located in 26 different departments or units on our campus.
Although an ideal candidate would have experience in confocal scanning laser microscopy, bright field microscopy, wide field fluorescence microscopy, deconvolution, image processing/analysis, all types of microtomy, immunocytochemistry, in situ hybridization and Adobe Photoshop, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Electron microscopy is not a component of this core facility. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired. This position has a salary range of $30,257 $54,663.00 and is 100% benefit eligible. Opportunities for professional development will be encouraged. Columbia is a great, inexpensive place to live.
The Core's web site can be viewed at http://www.biotech.missouri.edu/mbp/cores/index.html
Address applications (CV and 3 letters of reference) or inquires to:
Thomas E. Phillips, Ph.D. Division of Biological Sciences 3 Tucker Hall, University of Missouri Columbia, MO 65211-7400. 573-882-4712 PhillipsT-at-missouri.edu
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I agree in principal. But, we live in a very litigious, cover-your-ass type of world. It behooves employers and supervisors to document the training and warnings that women (and men) receive with respect to chemical hygiene in the laboratory environment -especially with respect to pregnancy. I would wager to bet that even the most idealist feminist would be looking for litigation relief if they were exposed to hazardous materials during their pregnancy even if it was her own fault with respect to educating herself to the dangers.
I think that people should be warned of the dangers and that warning should be documented. Their impending motherhood (or lack of) should be their own business. If a woman is pregnant and the unborn child can be potentially exposed to hazardous chemicals, the employer can only make adjustments to the woman's work environment if they are told. If they are not, they should not be held accountable. To protect the employer, there should also be documentation that determines when a woman first tells the company. Even when the training and documentation is there and the employee chooses to ignore them, a company can still be held responsible.
And another thing: this thread is about responsible scientists doing things to protect beings that can not protect themselves. They approached this problem in a responsible, caring and concerned way, not to subjugate women. It is nothing to get incensed about.
CYA rules.
These are just my humble opinions.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Kristen Lennon [mailto:kalen-at-iastate.edu] Sent: Monday, July 16, 2001 5:30 PM To: Microscopy-at-sparc5.microscopy.com
} To All following this lead, Is anyone concerned about the probability that the treatment of women described by several of the listers is illegal? Treating a woman like a criminal or a delinquent child who needs to be closely monitored as to her "motherhood" status is not only ridiculous, but descriminatory and hopefully illegal. It is the responsibility of each individual in a laboratory to watch out for his or her own safety and therefore the business and responsibility of each woman in the lab to watch out for the safety of her unborn child. We are all well past the kindergarten stage and should no longer require a hand to hold while crossing the street. Incensed in Iowa, Kristen
} to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified soldering } iron) has not worked well so far, any ideas how to eliminate xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
Can anyone recommend a vendor for a confocal microscope for materials science use? We would like to discuss a materials science application with a knowledgeable vendor..
Thanks, Steve Bradley UOP 50 E Algonquin Road Des Plaines, IL 60017 1-847-391-3321
This year we will be hosting the 7th Annual RMC Materials Microtomy Short Course in Tucson, Arizona from October 23 - 26.
For those of you residing in colder climes Tucson has just the weather to chase away the onset of those winter-time blues.
Join us, and our internationally renowned course faculty, in sunny Tucson to participate in this unique event. This short course is designed specifically for researchers in the field of materials science who wish to gain exposure to advances in specimen preparation for electron microscopy.
email stacy-at-boeckeler.com to receive full details and a course brochure
Dave Roberts Director - RMC EM Products Boeckeler Instruments, Inc 4650 South Butterfield Drive Tucson, AZ 85714 Tel: (520) 745-0001 Fax: (520) 745-0004 email: dave-at-boeckelr.com website: www.rmcproducts.com
I've registered for M & M 2001 in Long Beach, but I'm temporarily a single parent during the event, and need to make arrangements for my six-year-old child. One option is to bring her with me...but I need to make sure she is looked after during the short courses and some of the scientific symposia. If some local listers could give me any suggestions for care, or if there's anyone else in the same position who would be willing to pool expenses, I would be very grateful if you could contact me (you can e-mail me directly).
Thanks in advance,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
"I am currently evaluating EDS packages for my Philips XL-30 ESEM with LaB6 .................What about water vapor and ice build up? ....
I acquired a (Si) detector from Oxford for our ESEM 2020 in 1995. At that moment half a dozen detectors had been sold by Oxford to ESEM users, half a dozen by Noran, and one by EDAX. Oxford offered in its software package the possibility of "conditioning" the detector, a partial warm-up to remove ice etc., which takes two hours only. The other companies did not offer that possibility at that time, probably because it was patented. I do not know whether they offer it by now. It is very easy. I have used it for 7 years now, without problems so far. I believe that regular "conditioning" in combination with other measures has allowed this. Other measures: 1. a protective tube in which the detector is retracted when not operating (or during hot-stage operation, to protect from hot dust); and 2. maintaining a low pressure in the sample chamber (1 Torr instead of the regular 5 T), when the ESEM is not used.
As to : " How do members deal with the beam spread issues? " ,
X-ray mapping is a good method to eliminate the effects of beam spreading, and check particle analyses. Furthermore, beam spreading can be reduced by working at the lowest pressure possible without getting charging (I use 0.9-1 Torr routinely) and by using the detector set-up (the elongated detector/bullit in the ESEM 2020) with the shortest path in the higher-presssure surrounding, i.e. the sample chamber. I know that for quantitative analysis new software programmes exist which allow e.g. to extrapolate the results from a higher-pressure analysis + a lower-pressure analysis to zero pressure, or use some method of background/surroundings subtraction (several methods have been mentioned in the literature), but my (older) software does not offer that possibility. With best regards, Emond de Roever
We are a hospital EM Unit performing surgical pathology examinations and virology exams on clinical specimens. We perform EM on about 130 surgical cases per year (works out to about 150 specimens, nearly half of which are kidneys, the remainder are solid tissue tumors, muscle, nerve). We perform direct EM on about 300 stools/vesicle fluids for virus examination per year.
We have 1.4 FTEs (full-time equivalents). Our .4FTE covers leaves and off-sets workload. We are a manual service.
Our TAT for pathology is 3 working days from processing (day 1), thick sectioning/consult with pathologist/thin sectioning, staining/scoping (by technologist-specialist)/photography (day 2), photographic printing and reporting to pathologist (day 3).
Has anyone ever worked out an estimate of how many "man staff-hours" are involved in processing a biopsy specimen, manually (pardon me again!), for EM? This includes scoping, interpretation and reporting.
Thank you,
C. Powell EM Unit Saint John Regional Hospital NB, Canada powca-at-reg2.health.nb.ca
We have been critical point drying scallop specimens successfully for SEM to examine them as they grow. We are now at a point where the specimens no longer fit in the CPD chamber. With only a couple of specimens to go we thought to try HMDS. The samples are approximately 30mm in diameter. Does someone have a protocol they have used for larger specimens that has worked for them? You can send them directly to me and I can summarize for the list. Thanks in advance.
David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/micros_e.html
I have lost the e-mail address of Lesley Bechtold of Jackson Labs. Can some one help me out? Thanks, Tim
Timothy Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 238 JAH 1020 Locust Street 19107 215-513-4798 Work 610-613-8170 Cell
I can add a few points to this reply. Since you have the XL30, you will generally be leaving it in Hivac overnight and when not in use for ESEM. Ice buildup should be a lesser problem for you; depending of course on the nature of your work. I have the Oxford system on our XL30 ESEM-FEG and have had good results with the package and with the service (rarely needed). I did not purchase the package myself so did not evaluate competitors' offerings. I don't know which detectors you have but the lower-pressure ones are best for EDS: Large Field Detector or CP+ insert with Backscattered ED will operate at much less than 1T. If you need higher pressures; use the extended cone X-ray detectors. I have used the conditioner on occassion; I believe this has been a feature on Link systems since the early 90's at least. Monitor low-end sensitivity with NiLa vs. NiKa.
I find that beam skirt is a minor issue; generally the ESEM samples (customers) are not using EDS. When you need both, spot-check surrounding areas, or run a few maps at low resolution, and be less "quantitative" about your conclusions unless you need to get into the corrections mentioned by Emond.
The new Oxford system is significantly different from mine, so I won't talk about ease-of-use and portability of data.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Tuesday, July 17, 2001 3:48 AM, Emond W F de Roever [SMTP:ederoever-at-ONDEO-Nalco.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Scott, } } Responding to your question: } } "I am currently evaluating EDS packages for my Philips XL-30 ESEM with } LaB6 .................What about water vapor and ice } build up? .... } } I acquired a (Si) detector from Oxford for our ESEM 2020 in 1995. At that } moment half a dozen detectors had been sold by Oxford to ESEM users, half a } dozen by Noran, and one by EDAX. Oxford offered in its software package the } possibility of "conditioning" the detector, a partial warm-up to remove ice } etc., which takes two hours only. The other companies did not offer that } possibility at that time, probably because it was patented. I do not know } whether they offer it by now. It is very easy. I have used it for 7 years } now, without problems so far. I believe that regular "conditioning" in } combination with other measures has allowed this. Other measures: 1. a } protective tube in which the detector is retracted when not operating (or } during hot-stage operation, to protect from hot dust); and 2. maintaining a } low pressure in the sample chamber (1 Torr instead of the regular 5 T), } when the ESEM is not used. } } As to : " How do members deal with the beam spread issues? " , } } X-ray mapping is a good method to eliminate the effects of beam spreading, } and check particle analyses. Furthermore, beam spreading can be reduced by } working at the lowest pressure possible without getting charging (I use } 0.9-1 Torr routinely) and by using the detector set-up (the elongated } detector/bullit in the ESEM 2020) with the shortest path in the } higher-presssure surrounding, i.e. the sample chamber. I know that for } quantitative analysis new software programmes exist which allow e.g. to } extrapolate the results from a higher-pressure analysis + a lower-pressure } analysis to zero pressure, or use some method of background/surroundings } subtraction (several methods have been mentioned in the literature), but my } (older) software does not offer that possibility. With best regards, Emond } de Roever } } Ondeo-Nalco Europe, Oegstgeest, the Netherlands } }
On my three year old XL30 I usually work at 6 Torr (to keep specimens really wet) and working distance 7.5 mm. It is routine work to get decent maps or line scans of Ca, P, S (4-10% consentration). If you do not need trace elements or quantitative analysis, EDS on ESEM will do pretty good work.
If you samples are not wet, just not conductive, then you can go to higher voltage, lower pressure (1 Torr or less) and use BSE, which is less influenced by charging and then it will be difficalt to find difference with conventional high vacuum EDS analysis.
My detector have never had ice build up, but oil contamination became substantial in a few weeks after cleaning. As far as I know others do not have this particular problem, and I am loosing my battle with EDS and microscope manufacturers.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Scott Whittaker [mailto:Whittaker.scott-at-nmnh.si.edu] } Sent: Monday, July 16, 2001 1:16 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: xl_30 ESEM and new EDS } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } I am currently evaluating EDS packages for my Philips XL-30 ESEM with } LaB6 and wanted to poll the list members for their opinions, be they } positive or negative. How do members deal with the beam spread } issues? Can we trust the results? What about water vapor and ice } build up? As a multi-user facility how easy is it to get the data out } to the researcher these days? What about service quality? } } I will post a summary so if you prefer your responses excluded, } please let me know. } } Thanks } } Scott Whittaker } SEM Lab Manager } National Museum of Natural History } Smithsonian Institution } 10th & Constitution Ave, NW } Washington DC 20560-0104 } 202-357-1651 } } }
I seen several comments on this problem. It is a legal minefield. It is important for companies to get an expert attorney's advice on this matter. Several years ago a car battery manufacturer tried to limit the exposure of pregnant women to lead by re-assigning them to different jobs. The case went to the U.S. Supreme Court and the company lost. The Court said in an unanimous opinion that the company must make the workplace safe for all employees equally. A better approach is to give all employees, regardless of gender, the same safety education at the time of hiring and to periodically update this as needed. In other words men would also be told of the hazards for pregnant individuals. The best approach is to simply make the workplace safe for pregnant women. Get proper legal advice. This is not for amateurs.
I can ask around with some of our friends who watch our children to see if anyone would be interested in heading up to Long Beach. We are about a 1 hour drive from the convention center so I would guess they would want to do a whole day thing rather than just a few hours. If you think that would help, let me know and I will see what I can arrange.
Best regards-
David
Rick Powell at Nanoprobes wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Microscopists: } } I've registered for M & M 2001 in Long Beach, but I'm temporarily a single } parent during the event, and need to make arrangements for my six-year-old } child. One option is to bring her with me...but I need to make sure she is } looked after during the short courses and some of the scientific symposia. } If some local listers could give me any suggestions for care, or if there's } anyone else in the same position who would be willing to pool expenses, I } would be very grateful if you could contact me (you can e-mail me directly). } } Thanks in advance, } } Rick Powell } } ***************************************************************************************** } Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com } NANOPROBES, Incorporated } 95 Horse Block Road, Yaphank, NY 11980-9710, USA } } US Toll-free: (877) 447-6266 * Tel: (919) 845-6324 * Fax: (631) } 980-3608 } } Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html } *****************************************************************************************
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
Even though I am a female, I have to agree with Scott on this one. People will seize any opportunity to sue, no matter how great their understanding of the risks involved ahead of time - I have seen it myself in a lab situation. I would love to think that people would take responsibility for their own actions, but that just isn't the reality these days - the lure of easy money always ends up being too great. So, as Scott says, you CYA as best you can and keep your fingers crossed.
Jan Markham Motorola
Kristen Lennon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To All following this lead, } Is anyone concerned about the probability that the treatment of women } described by several of the listers is illegal? Treating a woman like a } criminal or a delinquent child who needs to be closely monitored as to her } "motherhood" status is not only ridiculous, but descriminatory and } hopefully illegal. It is the responsibility of each individual in a } laboratory to watch out for his or her own safety and therefore the } business and responsibility of each woman in the lab to watch out for the } safety of her unborn child. We are all well past the kindergarten stage and } should no longer require a hand to hold while crossing the street. } Incensed in Iowa, } Kristen } } } to all out there: } } } } This is what I have done so far: } } 1. Removed all stored chemicals from the lab. } } 2. Moved all chemical mixing into fume hood. } } 3. Met for 3 hours with environmental safety officer. } } 4. Met for 2 hours with radiation safety officer. } } 5. Eliminated all protocols which involve teratogens from pregnant } } workers routine. } } 6. Established a personal hygiene plan for pregnant worker, which is } } being signed of by worker, myself, director of the lab, safety } } officer, and her physician. } } 7. Reiterated safety protocols for worker, coat , gloves, safety } } glasses, etc. } } 8. Had lab monitored for radiation, passed. } } 9. Had lab monitored for formaldehyde gas, passed. } } 10. Planning to monitor xylene exposure to worker when thin sectioning } } (cannot do this in the hood, obviously). Heat pen( modified soldering } } iron) has not worked well so far, any ideas how to eliminate xylene? } } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } } LR White. Any ideas? } } I am experimenting with food related stains (grape juice, etc.). } } 12. Employee has completed, lab safety training course, formaldehyde } } training course and radiation training course. } } 13. Will continue to monitor lab for chemical and radation exposure. } } } } To some this may seem like over kill, but I believe that it is very } } important that every stone is turned in regards to safety. Another } } issue that I face is that I have numerous grad students coming through } } the lab, and there is always a possibility that they are pregnant and do } } not realize it or keep it from me. I also have a nursing mother who was } } visiting the lab and never bothered to tell me, until the subject of why } } the lab was reorganized came up. I now demand that all female student, } } techs, visitors, ect. } } tell me of their "motherhood" status. } } } } Bill } } } } -----Original Message----- } } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } } Sent: Tuesday, July 10, 2001 7:28 AM } } To: William McManus; Monson, Frederick C.; Sally Stowe } } Cc: Microscopy Listserver } } Subject: RE: Tol blue/borax } } } } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } } ----------------------------------------------------------------------- } } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------- } } Kristen A. Lennon, Ph.D. } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } 515-294-8854 } kalen-at-iastate.edu } www.baumlab.org
Other organizations have child care options. I believe that the AVS has had something available and the ICMCTF meeting has been talking about it. I have a colleague who used it at the AVS and I believe that she said it was on the expensive side, but that it did enable her to go to the meeting. I know of other colleagues who have not attended meetings because of conflicting schedules with their spouses.
Perhaps this is a topic that should be discussed at the MSA and MAS business meetings. Since my kids are grown, it doesn't affect me, but I can understand the problem.
Another option is to use a forum such as the listserver to organize a child care pool or to pool the resources of those members that need such a service.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Rick Powell at Nanoprobes [mailto:rpowell-at-nanoprobes.com] Sent: Tuesday, July 17, 2001 2:21 AM To: Microscopy-at-sparc5.microscopy.com
Hello Microscopists:
I've registered for M & M 2001 in Long Beach, but I'm temporarily a single parent during the event, and need to make arrangements for my six-year-old child. One option is to bring her with me...but I need to make sure she is looked after during the short courses and some of the scientific symposia. If some local listers could give me any suggestions for care, or if there's anyone else in the same position who would be willing to pool expenses, I would be very grateful if you could contact me (you can e-mail me directly).
Thanks in advance,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Along this vane, We became concerned about safety issues quite a number of years ago. We try to train users of this facility as to the dangers involved and appropriate scientific procedures...regardless of sex, age, etc. We have made it a point to provide sufficent hoods, etc so that users can work safely. We also make a point of the fact that it is the researcher's responsibility to inform themselves of hazards and to ask questions if they are not sure of appropriate procedure. Material safety data sheets are also available in an indexed binder so that anyone can easily check on specific reagents.
In addition we have them sign a sheet when they first start using the facility which says that they understand the potential risks (broad categories of possible hazards are listed on the sheet) and assume the responsibility for their own actions. This may not hold up completely in a court of law but it does show an attempt on our part to make the user aware of their own responsibility for their actions.
Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
I agree in principal. But, we live in a very litigious, cover-your-ass type of world. It behooves employers and supervisors to document the training and warnings that women (and men) receive with respect to chemical hygiene in the laboratory environment -especially with respect to pregnancy. I would wager to bet that even the most idealist feminist would be looking for litigation relief if they were exposed to hazardous materials during their pregnancy even if it was her own fault with respect to educating herself to the dangers.
I think that people should be warned of the dangers and that warning should be documented. Their impending motherhood (or lack of) should be their own business. If a woman is pregnant and the unborn child can be potentially exposed to hazardous chemicals, the employer can only make adjustments to the woman's work environment if they are told. If they are not, they should not be held accountable. To protect the employer, there should also be documentation that determines when a woman first tells the company. Even when the training and documentation is there and the employee chooses to ignore them, a company can still be held responsible.
And another thing: this thread is about responsible scientists doing things to protect beings that can not protect themselves. They approached this problem in a responsible, caring and concerned way, not to subjugate women. It is nothing to get incensed about.
CYA rules.
These are just my humble opinions.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Kristen Lennon [mailto:kalen-at-iastate.edu] Sent: Monday, July 16, 2001 5:30 PM To: Microscopy-at-sparc5.microscopy.com
} To All following this lead, Is anyone concerned about the probability that the treatment of women described by several of the listers is illegal? Treating a woman like a criminal or a delinquent child who needs to be closely monitored as to her "motherhood" status is not only ridiculous, but descriminatory and hopefully illegal. It is the responsibility of each individual in a laboratory to watch out for his or her own safety and therefore the business and responsibility of each woman in the lab to watch out for the safety of her unborn child. We are all well past the kindergarten stage and should no longer require a hand to hold while crossing the street. Incensed in Iowa, Kristen
} to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified soldering } iron) has not worked well so far, any ideas how to eliminate xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
I assume that you have tried other solvents as well, including water. You might consider adding a small surfactant.
One method that has worked for me for diamond particles is to ultrasonic a suspension of water and let it set for a few moments and then pour some of the liquid into a nebulizer. I then use the nebulizer over the surface that I want to disperse the diamond particles on while it is on a hot plate at about 120C.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Christopher M May [mailto:cmmay-at-US.ibm.com] Sent: Tuesday, July 17, 2001 12:52 PM To: microscopy-at-sparc5.microscopy.com
Hello,
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
Sometimes hotels have child care services or nannies who will come to rooms. Perhaps several folks could go in together and pool resources. Try calling your hotel concierge.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
} Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907
Hello- I am a grad student at the University of Idaho and would like some input on a protocol for fixing rat tissue in order to view their spinal ganglia on the TEM. First heparin is injected intracardiac, then perfusion via the heart with saline (37C) followed by paraformaldehyde (4%, 15C). The vertebral column is then removed and immersed in 4% paraformaldehyde for several hours before dissecting the spinal ganglia. This protocol was given to me by my advisor, but I have been unable to find a similar method cited in the literature. Also, I have used this method and found all the mitochondria have already lost their cristae. Any suggestions and comments would be appreciated. Thank you. --Meghan Towner
Of course, your technique will be selecting for the smaller particles as the larger ones as well as the agglomerates will be dropping out while it sets before you decant into the nebulizer. Nebulizing will also have issues about particle selection.
The original requester didn't say what the size range of the particles are but I assume that they must be submicron or nanometer range as he is using the TEM. Was he trying to size them, study their morphology, or what? I usually end up trying a variety of suspending fluids and also use on ocassion water with surfactant. But there are other techniques to use depending on what we're trying to find out about the particles.
Damian Neuberger Senior Research Scientist Particle Science and Technology Baxter Healthcare Corp
} I assume that you have tried other solvents as well, including water. You might consider adding a small surfactant. } } One method that has worked for me for diamond particles is to ultrasonic a suspension of water and let it set for a few moments and then pour some of the liquid into a nebulizer. I then use the nebulizer over the surface that I want to disperse the diamond particles on while it is on a hot plate at about 120C. } } -Scott }
Below is the result of your feedback form. It was submitted by (cokendolpher-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 17, 2001 at 19:33:06 ---------------------------------------------------------------------------
Email: cokendolpher-at-aol.com Name: James Cokendolpher
Organization: Midwestern State University
Education: Graduate College
Location: Lubbock, Texas USA
Question: I have used an AO150 for my home research for the past 20 or so years. With the advent of good digital cameras I am now thinking of buying an SL3D microscope and puting my camera lucida in a museum. Do you know of any other microscope with a greater depth of field for under $3000? Do you know of any articles that are good or bad about the SL3D; other than those on their website? My work involves the description and illustration of minute arthropod parts. Thanks.
Hi! I have a question regarding the removal of Spurrs resin from sections on slides. I want to use aniline blue as a callose stain for plant tissue. I have sections (embedded in Spurrs) on slides. The protocol I have to remove the resin uses sodium methoxide and benzene (Mayor, Hampton and Rosario, 1961). I was wondering if anyone can suggest a protocol that is less dangerous than using the sodium methoxide. Or, alternatively, has anyone used this technique, or a similar one and had any problems/success with it? Thanks, Rose
{center} {bold} {bigger} {bigger} ANALYTICAL GROUP {/center}
{center} ELECTRON MICROSCOPIST {/center}
{flushboth} {/bold} {smaller} {smaller} Applications are invited from suitably= qualified and experienced candidates for the above post. The successful candidate will be responsible for providing an electron microscopy service in support of the Institute's research programme and on a contract basis to commercial organisations. {/flushboth}
{flushboth} Candidates should possess a degree in an appropriate subject an= d be experienced in the analysis of a wide range of materials by scanning electron microscopy. {/flushboth}
{flushboth} Starting salary, depending upon qualifications and experience, will be normally within the range =A318,500 - =A321,400 per annum (performance related to a maximum of =A3 26,000). Non- contributory Superannuation Scheme. {/flushboth}
MLURI receives funding from the Scottish Executive Environment and Rural Affairs Department.
{flushboth} Further particulars and application forms can be obtained from Personnel, The Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen, AB15 8QH Tel 01224-498200, Fax 01224- 311556, E-mail { HYPERLINK mailto:e.cockburn-at-mluri.sari.ac.uk } {underline} = {color} {param} 0000,0000,FF00 {/param} e.cockburn-at-macaulay.ac.uk {/underline} {= color} {param} 0100,0100,0100 {/param} to whom completed application forms must be returned by 20 July 2001. {/flushboth}
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{center} {bold} {bigger} {bigger} ANALYTICAL GROUP {/center}
{center} ELECTRON MICROSCOPIST {/center}
{flushboth} {/bold} {smaller} {smaller} Applications are invited from suitably= qualified and experienced candidates for the above post. The successful candidate will be responsible for providing an electron microscopy service in support of the Institute's research programme and on a contract basis to commercial organisations. {/flushboth}
{flushboth} Candidates should possess a degree in an appropriate subject an= d be experienced in the analysis of a wide range of materials by scanning electron microscopy. {/flushboth}
{flushboth} Starting salary, depending upon qualifications and experience, will be normally within the range =A318,500 - =A321,400 per annum (performance related to a maximum of =A3 26,000). Non- contributory Superannuation Scheme. {/flushboth}
MLURI receives funding from the Scottish Executive Environment and Rural Affairs Department.
{flushboth} Further particulars and application forms can be obtained from Personnel, The Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen, AB15 8QH Tel 01224-498200, Fax 01224- 311556, E-mail { HYPERLINK mailto:e.cockburn-at-mluri.sari.ac.uk {FontFamily} = {param} Times New Roman {/param} } {underline} {color} {param} 0000,0000,FF00 {/pa= ram} {FontFamily} {param} Book Antiqua {/param} e.cockburn-at-macaulay.ac.uk {/unde= rline} {color} {param} 0100,0100,0100 {/param} to whom completed application forms must be returned by 20 July 2001. {/flushboth}
Quote Ref MA17/01
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Rose wrote: } Hi! } I have a question regarding the removal of Spurrs resin from sections on } slides. I want to use aniline blue as a callose stain for plant tissue. I } have sections (embedded in Spurrs) on slides. The protocol I have to } remove the resin uses sodium methoxide and benzene (Mayor, Hampton and } Rosario, 1961). I was wondering if anyone can suggest a protocol that is } less dangerous than using the sodium methoxide. Or, alternatively, has } anyone used this technique, or a similar one and had any problems/success } with it? } Thanks, } Rose
Rose, I use sodium ethoxy (a saturated NaOH solution in absolute ethanol) for the Spurr removal in plant tissues. Use 5 minutes for a semithin section with 0.35 micrometers. Wash shortly in ethanol and, after, distilled water. The technique gives good results and it does not use methanol (dangerous to your health). I think that the protocol can be used by you without problems. Dr. Rinaldo Pires dos Santos E-mail: rinaldop-at-uol.com.br Plant Anatomy Laboratory - Dept. of Botany UFRGS - Porto Alegre - RS Brazil
Applications are invited from suitably qualified and experienced candidates for the above post. The successful candidate will be responsible for providing an electron microscopy service in support of the Institute's research programme and on a contract basis to commercial organisations.
Candidates should possess a degree in an appropriate subject and be experienced in the analysis of a wide range of materials by scanning electron microscopy.
Starting salary, depending upon qualifications and experience, will be normally within the range £18,500 - £21,400 per annum (performance related to a maximum of £ 26,000). Non- contributory Superannuation Scheme.
MLURI receives funding from the Scottish Executive Environment and Rural Affairs Department.
Further particulars and application forms can be obtained from Personnel, The Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen, AB15 8QH Tel 01224-498200, Fax 01224- 311556, E-mail { HYPERLINK mailto:e.cockburn-at-mluri.sari.ac.uk }e.cockburn-at-macaulay.ac.uk to whom completed application forms must be returned by 20 July 2001. Quote Ref MA17/01 Web Site { HYPERLINK http://www.mluri.sari.ac.uk }http://www.macaulay.ac.uk
I am going to wade in on this, despite the potential for being said to be beating a dead horse:
During a graduate virology course a few years ago it became apparent that one of our students was pregnant. Out of concern for her and her child, we took special interest in providing information to her, and reducing her exposure to chemicals. We run a safe ship, but everyone should acknowledge that during gestation a developing child is vulnerable to things the mother may be exposed to. In this case, the students worked in pairs, and it was a simple matter the have her partner handle chemicals of concern. Since she was a new graduate student, we felt it was responsible to provide her with all the information we could for her safety and her child's. Her learning experience was not hindered, and on a personal level I think she appreciated our concern.
Information and education are critical to learning to make responsible decisions regarding safety, pregnant or not.
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, July 17, 2001 7:38 PM To: Debby Sherman Cc: microscopy-at-sparc5.microscopy.com
Hear hear!
} Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907
Your protocol is for immunoEM with perfusion-fixation. I think perfusion-fixation is necessary for your experiment. But if your purpose is to see the ultrastructure, you should use 2% glutaraldehyde in stead of 4% paraformaldehyde as the fixative for the primary fixation. Your protocol for the perfusion is okay. Just try gluta.
Greg Ning
Gang Ning, M.D., Ph.D. Director EM Facility Medical College of Wisconsin
Meghan Towner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello- } I am a grad student at the University of Idaho and would like some input on a protocol for fixing rat tissue in order to view their spinal ganglia on the TEM. First heparin is injected intracardiac, then perfusion via the heart with saline (37C) followed by paraformaldehyde (4%, 15C). The vertebral column is then removed and immersed in 4% paraformaldehyde for several hours before dissecting the spinal ganglia. This protocol was given to me by my advisor, but I have been unable to find a similar method cited in the literature. Also, I have used this method and found all the mitochondria have already lost their cristae. Any suggestions and comments would be appreciated. } Thank you. } --Meghan Towner
Your solution depends on what type of TiO2 you have and the particle size of the TiO2 you want to disperse. It can depend on whether you want to see ultimates, agglomerates or aggregates (primary structural units). You gave no indication of the type of TiO2 you want to disperse or if it was milled.
R900, 901, 100, &101 Dupont equivalents will have to be dispersed differently from something like micronized TiO2 (very agglomerated) or something like Nanotech TiO2.
Dispersion can depend on the TiO2 being rutile (I assume this is your type (coated)), anatase (uncoated), or a mixture of both, like Nanotech's spherical TiO2.
The technique we used for TiO2, and still use, was written up and given at the University of Michigan to the president of JEOL (Dr. Ito) and Prof. Wil Bigelow of the University of Michigan in the 1960s.
The presentation was given in the about 1963? by Robert Fear and was called A Versatile Mulling Technique (or Method) for Dispersing Fine Powders (or Pigments). Bob says it was sent to the EMSA / MSA educational archives. It was complete with colored 35mm slides. We have no idea where this material is currently at the MSA.
Paul Beauregard / Robert Fear Sr. Research Associate PPG Industries Monroeville Technical Center Monroeville, PA 15601
Any opinions are ours and not those of PPG Industries as a whole.
-----Original Message----- } From: Christopher M May [mailto:cmmay-at-US.ibm.com] Sent: Tuesday, July 17, 2001 12:52 PM To: microscopy-at-sparc5.microscopy.com
Hello,
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
Good morning, I am planning to complete the microscopical characterization of some selected waxes using a hot stage on a light mcroscope. There does not seem to be much information in the literature on this subject and I thought there must be some microscopists well acqainted with this topic. We have completed DSC thermograms and other than looking for melting/crystallization temperatures, degree of crystallinity, is there anything else to be characterized? Can anyone provide further guidance? Thanks.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Readers, A question many may be asking, how do I get from the L.A. airport to Long Beach? I am advised that there is a (blue) bus system called SuperShuttle which leaves from outside the airline locations to Long Beach. Cost is $15 a head and it takes around a half hour and, typically, makes 3 stops. The taxi fare is around $45. Don Grimes, Microscopy Today
I don't understand Greg's comment (below). Power DTSA v 2.5.1, with the MSA 1.0 plug-in, will happily read EDAX Phoenix MSA files, which have a single column of spectral data as integers, and Link ISIS MSA files, which have the Y-data in 5 columns of reals. There is a tag, #DATATYPE, which, if set to "Y", tells the system to expect just the y-data, and not data pairs.
The problem with MSA files is that the MSA file specification is not a "format" - i.e., two people writing the same data to an MSA file can end up with different files, each of which is completely according to the specification. The problem for the reader is to be able to handle all the different possibilities.
One way that you can sometimes use to force a file to read in is to use the target system to write an MSA file (which, presumably, it will read back in) using the same channel specifiacations (number of channels, range, etc) as the source spectrum. Then use a text editor to copy the header from the target spectrum to the source spectrum. Tags like the #NCOLUMNS tag can be edited as required. Even this doesn't work if the format of the data itself is different, or if the target system will only accept a limited range of options (e.g. #NPOINTS must be a power of 2 in some systems), and the source doesn't match the required option.
The MSA reader should, of course, at least exit gracefully if it can't accept a spectrum, but not all do!
Tony Garratt-Reed.
} } The NORAN EMSA files follow the specifications fully and support all } varieties of EMSA options available. The problem is that DTSA only reads } one type of EMSA file format (X,Y pairs of data) which is not the default } condition of the VOYAGER files. The default format for VOYAGER stores only } the channel data in 5 columns. The default file format for VANTAGE was } changed to be X,Y pairs which is consistant with DTSA. } } There is a solution for VOYAGER. You can save the EMSA format compatible } with DTSA by using the shell command: } emsa_io -x=1 -w={path to store to} -s={spectrum name to store} } example: emsa_io -x=1 -w=$HOME/spectra/myspec.emsa -s=spectrum1 } } When the next version of VOYAGER software is released the default condition } for the emsa_io program will be converted to the same as VANTAGE which will } make VOYAGER's default version of EMSA file format consistant with DTSA. } } Greg Fritz }
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Hi Meghan, Try almost any of the introductory books on EM by Hayat, M.A.. Your library or advisor, or EM book shelf should have a copy. Most problems such as you describe are due to the failure to initiate fixation in a timely manner, the osmolarity of the vehicle, and the postfixation time and component sequence. Most in situ, perfusion fixations include a postfix with glutaraldehyde following the initial stabilization with HCHO (made FROM paraformaldehyde). Also, it is my experience that one can 'freeze' thin muscle bands with 4% HCHO at 4oC in 10 min. But one cannot satisfactorily FIX the material for subsequent light(via paraffin) or EM (via plastic) viewing in that time. Hayat appears to suggest 24hr postfix but also provides a 4hr fix to block technique, so almost anything, properly designed, can work. Hayat, in a small methods book (HAYAT, M.A., 1972. BASIC ELECTRON MICROSCOPY TECHNIQUES, VAN NOSTRAND REINHOLD, NEW YORK AND LONDON.) references a method by: [Schultz, RL and Case, NM (1970), "A modified aldehyde perfusion technique for preventing certain artifacts in electron microscopy of the central nervous system", J Microscopy, 92:69.] If you can't find either the book or the article, ask and I'll send the method (not brief!)
Good luck,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Meghan Towner } Sent: Tuesday, July 17, 2001 7:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: tissue preparation for TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello- } I am a grad student at the University of Idaho and would like some input } on a protocol for fixing rat tissue in order to view their spinal ganglia } on the TEM. First heparin is injected intracardiac, then perfusion via the } heart with saline (37C) followed by paraformaldehyde (4%, 15C). The } vertebral column is then removed and immersed in 4% paraformaldehyde for } several hours before dissecting the spinal ganglia. This protocol was } given to me by my advisor, but I have been unable to find a similar method } cited in the literature. Also, I have used this method and found all the } mitochondria have already lost their cristae. Any suggestions and comments } would be appreciated. } Thank you. } --Meghan Towner } }
After fooling with EELS data stored in EMSA format from a Gatan system for a while that was stored in rows of five of the Y-only format and trying to extract the data into Excel, I wrote my visual basic program to extract the data and then write the data back in x,y pairs so that I could very easily plot it in Excel. Then I went nuts in Visual Basic 6.0 and wrote the program to plot the data and play around with it. I mentioned it before; I will bring to M&M and make public.
Before I wrote the program, I tried to extract the data with the "rows of five" problem in Excel using macro programming, but gave up. The easiest way that I found to convert the data to single column data and then add the x data was to first use a word processor and get rid of the hard returns using search and replace and add the appropriate character to match the data separator in the row. I then removed the data separators and added a hard return. Then you can save as a text file and open the data file in Excel. You then parse the data correctly and move the column of data over by one. Then you use a series fill to generate the x data. You can then plot the data using Excel (or any other plotting program) to plot the x,y pairs.
If you want to, you can easily change the data headers in the EMSA data file to reflect the change in the way the data is now stored in the file. You can go to the EMMPDL library and find the file with the standard and figure out how to do it relatively easily.
I think that the biggest problem with the EMSA format is allowing the Y-only data to exist in rows of n values. However, I understand why that was done. For the older programming languages, it was easy to format the read/write statement to bring the data in/out that way. However, it is not a major stumbling block.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Anthony J. Garratt-Reed [mailto:tonygr-at-mit.edu] Sent: Wednesday, July 18, 2001 11:32 AM To: Fritz, Greg; microscopy-at-sparc5.microscopy.com
I don't understand Greg's comment (below). Power DTSA v 2.5.1, with the MSA 1.0 plug-in, will happily read EDAX Phoenix MSA files, which have a single column of spectral data as integers, and Link ISIS MSA files, which have the Y-data in 5 columns of reals. There is a tag, #DATATYPE, which, if set to "Y", tells the system to expect just the y-data, and not data pairs.
The problem with MSA files is that the MSA file specification is not a "format" - i.e., two people writing the same data to an MSA file can end up with different files, each of which is completely according to the specification. The problem for the reader is to be able to handle all the different possibilities.
One way that you can sometimes use to force a file to read in is to use the target system to write an MSA file (which, presumably, it will read back in) using the same channel specifiacations (number of channels, range, etc) as the source spectrum. Then use a text editor to copy the header from the target spectrum to the source spectrum. Tags like the #NCOLUMNS tag can be edited as required. Even this doesn't work if the format of the data itself is different, or if the target system will only accept a limited range of options (e.g. #NPOINTS must be a power of 2 in some systems), and the source doesn't match the required option.
The MSA reader should, of course, at least exit gracefully if it can't accept a spectrum, but not all do!
Tony Garratt-Reed.
} } The NORAN EMSA files follow the specifications fully and support all } varieties of EMSA options available. The problem is that DTSA only reads } one type of EMSA file format (X,Y pairs of data) which is not the default } condition of the VOYAGER files. The default format for VOYAGER stores only } the channel data in 5 columns. The default file format for VANTAGE was } changed to be X,Y pairs which is consistant with DTSA. } } There is a solution for VOYAGER. You can save the EMSA format compatible } with DTSA by using the shell command: } emsa_io -x=1 -w={path to store to} -s={spectrum name to store} } example: emsa_io -x=1 -w=$HOME/spectra/myspec.emsa -s=spectrum1 } } When the next version of VOYAGER software is released the default condition } for the emsa_io program will be converted to the same as VANTAGE which will } make VOYAGER's default version of EMSA file format consistant with DTSA. } } Greg Fritz }
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Just a brief note to inform the microscopy community of the death of Dr. Lonnie Russell on July 11. Dr. Russell drowned while swimming in the ocean off the coast of Brazil. Lonnie made significant contributions in microscopy (both light and electron) and was an internationally recognized expert in the area of reproductive biology. I have more information about Lonnie, so please contact me personally and I will forward it to you.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
} The problem with MSA files is that the MSA file specification is not a } "format" - i.e., two people writing the same data to an MSA file can end up } with different files, each of which is completely according to the } specification. The problem for the reader is to be able to handle all the } different possibilities. }
The problem we have in implementing a robust MSA file reader is lack of example files from the various MSA file writers. Because there are several flavors in implimenting the writer, many programmers take different paths which should not make a difference to a MSA file reader. When programming the MSA file writer, one sometimes does not think of the other possibilities. It's only when users complain that they cannot read a MSA file from some other system that the problems become known. Then resources have to be allocated to fix a problem that should have been found during the testing phase of the software. Does not seem to be hard but this can be costly (from a company viewpoint).
Something that would help this oversight would be a repository for the various MSA file flavors, something one could download to use as a test case for the robustness of their MSA file reader. The spectral content could be anything but should be real (AlCu calibration for example).
We would be happy to sponser such a location on our web site.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I thought I would forward an example of the kind of madness to which one may be driven by the need to avoid liability in the workplace. The gentleman who formulated the list at the base of this forwarded email had earlier requested information about how others handled pregnancy among workers in electron microscopy laboratories. He then reported his solution as you can see below.
I am inserting without comment a statement from our lawyers, Dumb, Dumber and Dumbest, Esquires, LTD., LLD, LLC, LLB, LLA, LZ, LY, which they returned in response to a question about my potential liability on this matter.
"Understanding that most exposures to hazard occur as a consequence of accident or personal negligence, we have come to the conclusion that it is impossible, if not illegal, to protect anyone in particular, much less someone who is specifically pregnant, from exposure to hazardous materials, especially if the individual is going to take a job that involves such hazard and continue to do THAT job during pregnancy. Thus, it behooves the employer to remove such hazards completely or to reduce them for those at greatest risk so that there is no risk to all, especially those who periodically enter periods of existence during which their risk increases. However, we must state that for us this is a matter of humane treatment of subordinates, especially those weak in mind if not in body, not the law, which in some measure is inscrutable, even to us.
The legal intricacies of this matter in particular border on the insane, and force us to adopt unusual and novel measures and reasoning to protect both the employer and its employees. It appears that the Supreme Court in one of its more enlightened decisions, ruled that a company may NOT discriminate among its employees in the matter of hazardous substance risk. Thus, a battery manufacturer was not permitted to protect pregnant employees by transfer away from lead, when it did not similarly protect the remainder of the work force. In fact, this ruling is quite reasonable given the fact that during the first trimester of pregnancy, the Supreme Court had already ruled that pregnancy, especially during the period mentioned, is in NO respect the business of Society (nor, by any extension, management).
Therefore, if during the first trimester, an accident might occur to damage that which is none of Society's business, there is no recourse set aside for management that can protect that which is none of ITS business during that period. We can all, however, assure the employer that should a pregnant employee decide to continue a pregnancy beyond the first trimester, the employer becomes wholly responsible for any damage that occurred to the object of the pregnancy during the time that the object of the pregnancy was none of its business. It is a quirk of this situation that the object of the pregnancy has no legal standing either during this period, when it is none of Society's or management's business, or, for that matter, during the entire duration of pregnancy. Thus, the employer is uniquely at risk of inflicting damage on an object with no legal standing during a period when that object is none of the employer's business and, legally, does not exist. Of course, one might argue, just as logically, that it is not possible to inflict harm on something that does not exist, but that avoids the practical reality. We continue to expand this reasoning for the purposes of expected appeals should our position prove, in the first instance, legally unreasonable (charges, as stipulated, per hour). The only recourse may be a standard agreement to be signed by ALL employees, that ANY who become pregnant (through physical or ethereal means, regardless of anatomy or lack thereof exhibited by themselves or others related or unrelated to the acts involved or uninvolved, philosophy, political party, voting record, religion, non-religion, sexual disposition or preference, or sexual non-preference or non-disposition, or any absence or presence of preference or disposition and notwithstanding the expected or unexpected effects of genetic predisposition or specific incapacity or capacity) will agree to be transferred to non-hazardous, riskless, activities for the course of that pregnancy, nominally nine months from the moment of conception. If this solution is adopted by the employer, then the employer can be assured that such actions will protect those offspring of employees who have or will have legal standing and for whose well-being the employer can ever be held responsible, if someone decides that such well-being was or is the employer's business.
Since discrimination against the parent is not permitted, it is obvious that all individual distinctions must be suppressed in any safety considerations and actions. For those who refuse such PROTECTION during pregnancy, patently hazardous employment should not be available at any time during which pregnancy (through physical or(see above)....), may or might be improbably possible. The other side of the coin is that all employees, regardless of job, must be treated identically. Thus once safety considerations have been installed in equal consideration of all, all employees will be able and free to enter, with equanimity, their job space by first travelling through those non-discriminating, hazardous, but equally safe and non-hazardous work spaces.
The reasoning for the above solution is simple and straight forward. Since there is no absolute evidence to suggest that true hermaphroditism is not possible in humans, it is reasonable for the employer to subject ALL employees to the above non-discriminatory employment process during the pre-employment interview as well as after employment. The employer, thus, has assumed the responsibility to dictate what it considers hazardous and potentially harmful to this broad group of individual employees. This is reasonable since the employer is at risk for damage to the potential offspring during times when the potentially pregnant employee may bring the potential offspring to the place of employment when such pregnancy is none of the employer's business (but when the employer's potential risk is real). It is not that the employer has rights nor can be said to have responsibility during the time of pregnancy when such pregnancy is none of its business, but the employer can claim the right to protect itself, when the potential offspring, during the time that when its existence is none of the employer's business, is exposed to hazards on the employer's property that may cause harm to the potential offspring when the parent decides to continue the pregnancy beyond that period when it is none of Society's or any employer's business, such that the potential offspring is permitted to continue to develop in such manner that it can potentially claim rights of redress for harm done during that time of its non-existence when it was exposed to hazard when such exposure, by simple logic, was none of the employer's business, and concurrently could not have happened."
It should be clear that the solution recommended by our legal team treats all potential employees in the same potential manner, giving all their potential rights according to Supreme Court ruling and protecting all equally without potential discrimination. Thus, reason, not chaos, will prevail!
Please pardon any errors in the above. I have not had time to review it since it was returned, a second time, edited for clarity and brevity, from the lawyer's office.
Since I cannot, in good conscience, ignore the above reality, I am obliged to pass this information on to you, my superiors, who will also be forced to become 'part of the problem' rather than 'part of the solution'. My apologies to all - indiscriminately! To those pregnant employees who insist that hazardous work must continue without hazard and, thus, constrain their employers to liquidate the workplace, I accord you the sensitivity and concern that I normally reserve for the incoherent. I may wonder what you are saying, but I never try to understand what you are thinking.
Fred Monson, who wouldn't suffer his wife, and thus, their unborn children, to work in the same business as he, but whose daughter has freely chosen it.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: William McManus } Sent: Tuesday, July 10, 2001 11:30 AM } To: Leona Cohen-Gould; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: lab pregnancy } } to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified } soldering } iron) has not worked well so far, any ideas how to eliminate } xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } . } } } } } } The issue of pregnant (or possibly pregnant) workers in the lab has } } turned out to be fairly complicated. If anyone else out there has beem } } through this situation, I would like to know how you dealt with it. } } } } Bill } } } } William McManus } } Supervisor } } Electron Microscopy Facility } } Department of Biology } } Utah State University } } Logan UT 84322-5305 } } } **************** } to Bill & All, } } When I was pregnant, 9 years ago, I was the entire staff of the EM } facility here (still was until a month ago)and so could not avoid } things. The way I handled safety concerns was to be extra cautious: } I worked in the hood, wore double gloves when working with fixatives } & resins and gloves for other times when i would not usually wear } them( like using Tol Blue), safety glasses if splashing was a } possibility, washed my hands almost compulsively. In short, I used } common sense. I was, and am, lucky that (UrAc aside) I don't have to } deal with radioactivity. colleagues of mine who were also pregnant } at the time (we had a rash of pregnancies that year) who did need to } work with isotopes minimized their use, got new shields, and at times } convinced labmates to do those steps for them! } } We all, thankfully had healthy babies who are now all approaching } adolescence and driving us nuts, but that's another issue for another } forum! } } Lee } } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } }
Scott, I have a couple of them on my disk and that is included in my program distribution files. I am going to put it in the MSA and MAS libraries. I am bringing it to M&M and I will give you a copy. They include Gatan, Noran, Emispec, and one other that I can't think of at the moment (I think that it was Oxford).
I had asked for examples and I got a few to try in the program.
If anyone would like to send me one of their EMSA formatted ASCII output EDS or EELS spectra files and tell me the vendor associated with source, I could include them with the distribution files.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Scott D. Davilla [mailto:davilla-at-4pi.com] Sent: Wednesday, July 18, 2001 2:25 PM To: Microscopy (E-mail)
} The problem with MSA files is that the MSA file specification is not a } "format" - i.e., two people writing the same data to an MSA file can end up } with different files, each of which is completely according to the } specification. The problem for the reader is to be able to handle all the } different possibilities. }
The problem we have in implementing a robust MSA file reader is lack of example files from the various MSA file writers. Because there are several flavors in implimenting the writer, many programmers take different paths which should not make a difference to a MSA file reader. When programming the MSA file writer, one sometimes does not think of the other possibilities. It's only when users complain that they cannot read a MSA file from some other system that the problems become known. Then resources have to be allocated to fix a problem that should have been found during the testing phase of the software. Does not seem to be hard but this can be costly (from a company viewpoint).
Something that would help this oversight would be a repository for the various MSA file flavors, something one could download to use as a test case for the robustness of their MSA file reader. The spectral content could be anything but should be real (AlCu calibration for example).
We would be happy to sponser such a location on our web site.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
} Meghan Towner wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hello- } } I am a grad student at the University of Idaho and would like some input on a protocol for fixing rat tissue in order to view their spinal ganglia on the TEM. First heparin is injected intracardiac, then perfusion via the heart with saline (37C) followed by paraformaldehyde (4%, 15C). The vertebral column is then removed and immersed in 4% paraformaldehyde for several hours before dissecting the spinal ganglia. This protocol was given to me by my advisor, but I have been unable to find a similar method cited in the literature. Also, I have used this method and found all the mitochondria have already lost their cristae. Any suggestions and comments would be appreciated. } } Thank you. } } --Meghan Towner
If you want good ultrastructure use gluatarladehyde in the fixative, as little as 1% will make a significant improvement but 2-3% is more common. The buffer used is also important, you did not specify which you are using. Paraformaldehyed alone can give good ultrastructure but you will need to fix for several days at a minimum. If you want to do immunostaining, you may have to skip the glut. and suffer with less than optimal ultrastructure or risk killing off the immunoreactivity. It will depend on the antigen in question. Also, the temperature shift from 37 degrees C for the washout to 15 degrees C for the fixative is likely to induce vasospasm, reducing the amount of fixative delivered to the tissue. Do everything at one temperature, room temperature should be fine. A good basic EM text will be invaluable. Brenda Weakley's "Beginners handbook in biological transmission electron microscopy" is excellent. "Biological techniques in electron microscopy" by Clinton Dawes is also very good but it may be out of print.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
} Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. } Debby
While Debby's basic premise is correct, it is also a fact that a fetus can be adversely affected by chemicals, or levels of chemicals, that would pose absolutely no threat to adults. I have worked in situations where implementation of all the precautions necessary to prevent such fetal exposure would have made the job impossible. We acknowledge these differences when approving new drugs for the market, and restrict their usage accordingly. Is the lab situation so very different?
In laboratories where large numbers of EM micrographs are produced over the years and researchers may come and go over time, what system do you use to keep track of the micrographs?
We were thinking of implementing a computer database where information about each micrograph (such as date, microscopist, sample, conditions, location of the archived micrograph, etc.) can be stored for future reference. Is there any existing database software (preferably Macintosh based) for this purpose or any opinions on such packages?
Thanks for your help.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
Try Thumbsplus from Cerious software. (www.cerious.com) PC based. I don't know if they ever came out with the Mac based version, but they did have a beta version at one time.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Dennis C. Winkler [mailto:Dennis_Winkler-at-nih.gov] Sent: Wednesday, July 18, 2001 5:03 PM To: Microscopy-at-sparc5.microscopy.com
Good Day All!
In laboratories where large numbers of EM micrographs are produced over the years and researchers may come and go over time, what system do you use to keep track of the micrographs?
We were thinking of implementing a computer database where information about each micrograph (such as date, microscopist, sample, conditions, location of the archived micrograph, etc.) can be stored for future reference. Is there any existing database software (preferably Macintosh based) for this purpose or any opinions on such packages?
Thanks for your help.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
Thanks for your input, but let me clarify. I am working with Nanotech TiO2, I am looking at sizes 100nm and smaller. The particles are spherical and what I would eventually like are measurements of their diameters which reflects as accurate a distribution as I can get. I do not want aggolmerates, just the particles. Therefore, any technique that has a selection bias I would rather not use.
Thanks again, Christopher May IBM Analytical Services
----- Forwarded by Christopher M May/Poughkeepsie/IBM on 07/18/2001 11:38 AM -----
"Christopher M May" To: microscopy-at-sparc5.microscopy.com {cmmay-at-US.ibm. cc: com} Subject: Titanium Particle dispersion for TEM
07/17/2001 12:51 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
My name is Professor Sabel. Whoever you are, your email is being automatically sent to my account. I think you have a big problem somewhere. Please check.
please find below the announcement for a long-term EELS post-doc position at the group of Electron Microscopy for Materials
Science (EMAT) in Antwerp, Belgium http://www.ruca.ua.ac.be/emat/. May I ask you to distribute this to colleagues that may be interested.
Many thanks,
Nick Schryvers
EELS Post-doc position at EMAT, Antwerp, Belgium
A new post-doctoral position will be available as of January 1, 2002, at
the laboratory of Electron Microscopy for Materials Science (EMAT) in Antwerp, Belgium. The emphasis of the work will be on the combination of
EELS with HRTEM, STEM and EFTEM with the aim to develop advanced detection, processing and interpretation procedures yielding improved resolutions. The work will be performed in the framework of a joint program including a research group on image processing (VISIE) and one specialised in spectral analysis techniques (MiTAC), both from the same university. The ideal candidate should have an extensive experience in performing high precision EELS experiments in the TEM. The experiments will be conducted with two EELS systems (GIF 200(0)), one installed on a
CM30 FEG Ultratwin, designed for ultimate resolution of 0.1nm (using Focus Variation) and another one on a 3000F JEOL FEG machine equipped with a STEM unit with a probe size of 0.4 nm, EDX and FasTEM. At first model systems in the field of CMR and superconducting materials, semiconductor precipitates and diamond like carbon films will be investigated, but the aim is for generic principles useful for a large majority of materials. The position is in principle for one year, but can be extended by one year periods up to a total of 4 years. Salary is arranged following European and local standards and includes the full range of social security benefits (health care, retirement, holiday, …). Actual figures depend on proven experience of the candidate. Please send your extensive resumé to
Prof. Dr. Dominique Schryvers Senior Lecturer Electron Microscopy for Materials Research (EMAT) Department of Physics University of Antwerp, RUCA Groenenborgerlaan 171 B-2020 Antwerp, Belgium Tel: 32-3-2180247 Fax: 32-3-2180257 E-mail: schryver-at-ruca.ua.ac.be, nick.schryvers-at-ua.ac.be
ThumbsPlus is good if you have a digital image to keep track of, but I am not sure if Dennis was referring to that or to physical micrographs. If it is the latter then I would suggest using Microsoft Access and setting up their own forms and reports to suit their particular needs. Greg Erdos
At 05:32 PM 7/18/2001 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
Can anyone advise on removing a coverslip mounted using DPX? I am currently soaking it in xylene, but overnight didn't "cut the mustard"! Any cogent comments? Please?!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think the key here is (from the post below):
"No one should be allowed to expose themselves or others to dangers and it is THEIR RESPONSIBILITY to provide data to their managers if such a case exists or might exist."
If someone makes a medical condition known, ANY medical condition, to their supervisor, then it seems that the supervisor should take appropiraite additional action OVER AND ABOVE THE ROUTINE SAFETY PRECAUSTIONS to provide additional information and safety as required.
However, the question is not if we should attempt to make an environment as safe as possible (that does depend on specific individual need and may or may not be sufficiently possible in all situations) but whether a supervisor has the right to demand medical information from someone who may not be willing to share it. Do we have the right to not only demand the information but, upon hearing it, require the employee to observe restrictions when they, after being fully informed of hazards, etc, do not wish to do so.
Also, although this thread has dwelt on pregnancy as the issue of concern, it does take two to tango and concern for healthy sperm is as important as healthy eggs. Do we really know effects, limits, etc on all potential hazards in our labs to these tissues as well as to fetuses?
We can discuss this all day but, not being legal professionals, our thoughts are not necessarily sufficient in a court of law. I think it behooves all of us to provide as safe an environment as possible within reason and then respond to special situations in a way that is agreeable to all parties involved. This may require restrictions or reassignment of personnal if the environment cannot be made as safe an an employee/employer feels necessary (which will no doubt raise all kinds of additional questions/concerns). We are our brother's keepers only to the extent that they want to be kept!
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
Dennis,
You've probably already seen the 'Access' part of the MS Office programs as a database, but if not, I use it to keep track of the admin things in our microscopy unit and it works well, has lots of features. Another program that could be useful, although not exactly what you're looking for, is IrfanView, which lets you open up fast all the images you have in a folder, like a contact print of photos - later you can do things like copy, delete etc. on the files. It's freeware and you can download at
Both of these I've found really useful in general organization of the data and files that end up accumulating everywhere.
Good luck!
Mark
******************************************** Mark West, Encargado, (Manager) Unidad de Microscopia(Microscopy Unit), Lab, 301-Sur, Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, C.P. 04510, Mexico D.F., MEXICO
Pagina de Web (Web page) http://ifcsun1.ifisiol.unam.mx/microscopy.html ********************************************
On Wed, 18 Jul 2001, Dennis C. Winkler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Good Day All! } } In laboratories where large numbers of EM micrographs are produced } over the years and researchers may come and go over time, what system } do you use to keep track of the micrographs? } } We were thinking of implementing a computer database where } information about each micrograph (such as date, microscopist, } sample, conditions, location of the archived micrograph, etc.) can be } stored for future reference. Is there any existing database software } (preferably Macintosh based) for this purpose or any opinions on such } packages? } } Thanks for your help. } } - Dennis } } ------------------------------------------------------------------------ } Dennis C. Winkler, PhD. Phone: (301) 496-0131 } Laboratory of Structural Biology Research Fax: (301) 480-7629 } NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov } Bldg. 6, Room B2-26, MSC-2717 } Bethesda, MD 20892-2717, U.S.A. }
I have a JEOL 6320 FEGSEM equipped with a Oxford CT1500 cryo unit which I have used for a couple of years. I recently replaced the Cr foil/al back plate target with a Cr sputtered on aluminum target. Now I get a plasma but no deposition of Cr. (And yes, the target is facing the right direction). Yesterday, I replaced a leaky argon supply valve and metering valve. Still no deposition. I have also tried a full range of current/back pressure settings with no luck. Anyone have any ideas as to what the problem is?
The work is piling up so I am also looking for access to a FEGSEM with a CT1500 or CT2500 cryo unit equipped with a Cr target to get some high priority / high resolution work done. (Preferably somewhere here in the mid-west). Please contact me with the appropriate information.
Thanks, Dave Williams The Dow Chemical Company Midland, MI dmwilliams-at-dow.com
Thanks for all the responses to my search for child care options in Long Beach. For those who wanted to know (and anyone else who is also interested), here's what I found:
The Westin can arrange babystitting, estimated $ 15 - $ 20 per hour.
The Growing Years (1 World Trade Ctr # 199) was recommended. It has been re-named, but can look after children up to 5, telephone (562) 495-6300 (unfortunately not good for me - my child is 6)
Affordable Nannies & Care Givers, (562) 988-6588, is an agency which provides babysitting; usual charges $ 8 - $ 10 per hour plus fee of $ 25 per day.
If you are YMCA members, you can sign up for a few days camp at the YMCA, 1720 N Bellflower Blvd, (562) 96-3394 - it's called 'Sunshine Camp" and costs $ 105 for 1 to 3 days, $ 140 for 4 or 5 days, plus $ 35 registration fee; you need to go there first (i.e. Saturday or earlier) with vaccination records etc.
Using switchboard (www.switchboard.com) to search for businesses (select "child care services" from the list) gave me about 65 hits. One or two day care businesses said they could provide a few days' care, but most only go up to age 5.
Thanks again,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
Project MICRO (Microscopy In Curriculum - Research Outreach) is MSA's middle school educational outreach program. MICRO frequently sponsors programming at the annual M&M meeting - but because it doesn't fit the basic symposium structure, sometimes it "falls thru the cracks" in program preparation. That happened this year; the MICRO workshop isn't listed in the daily program. Although it's primarily intended for local teachers, it may interest you too.
Repairing School Microscopes: A Workshop for Teachers and Volunteers Tuesday, August 7 10 am - 3 pm, room 203C Led by David Bentley, University of Arizona
If you ask teachers what kind of help they need, a common response is "fix our microscopes"! In this context, "repair" means basic maintenance, rather than things requiring expert attention, and Dave has considerable experience in teaching the subject to teachers. If you want to help with educational outreach but you don't want to be a classroom volunteer, come to this workshop and learn how to help teachers in your local schools.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
We have inherited a Leitz Metallux 3 microscope with epifluorescence, however, without a user's manual (not a complaint, just a fact) and are wondering whether anyone knows a source for old microscope manuals. There are some quirky things about it, for example, the transmitted light is either yellow or red, never white, and, other than dismantling the entire body, and physically removing the filters, I cannot find a way to fix that. I could take it apart but I'm sure there would be parts left over after reassembly.
Thanks for any help.
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
OK, here you go -- SOP of TEM drop grid preparation for 100nm down or up TiO2 particles:
1) Dilute TiO2 solution to about 1% with DIW or other solvents. I have the luck to use straight H2O in most of the cases involving particles including TiO2. Surfactants sometimes cause more problems than resolve particles. Alcohol or acetone could be good solvents too 2) Sonicate the solution 3) Use a pair of negative tweezers, and pick up a 200 mesh Cu C-film grid WITH Cu-SIDE UP. You should catch as less edge of the Cu grid as possible. 4) Lay the tweezers on a table and place a quadrant of filter paper underneath the grid/tweezes. 5) Use a pipette to transfer a drop of the liquid onto the grid. Wick away extra liquid with half of that quadrant of the filter paper underneath the tweezes 6) Let it dry naturally or put under a light 7) Check with TEM 8) Later, experience will tell you how much concentration to use and how much liquid to drop onto the grid
********************************* Chaoying Ni Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
On Wed, 18 Jul 2001, Christopher M May wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } } Thanks for your input, but let me clarify. I am working with Nanotech } TiO2, I am looking at sizes 100nm and smaller. The particles are spherical } and what I would eventually like are measurements of their diameters which } reflects as accurate a distribution as I can get. I do not want } aggolmerates, just the particles. Therefore, any technique that has a } selection bias I would rather not use. } } Thanks again, } Christopher May } IBM Analytical Services } } } ----- Forwarded by Christopher M May/Poughkeepsie/IBM on 07/18/2001 11:38 } AM ----- } } "Christopher M } May" To: microscopy-at-sparc5.microscopy.com } {cmmay-at-US.ibm. cc: } com} Subject: Titanium Particle dispersion for TEM } } 07/17/2001 } 12:51 PM } } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I am working with a TEM and I would like to know the best way to disperse } Titanium Oxide particles for analysis. } I am currently using a methanol solution, and it is not working very well. } } } Sincerely, } } Christopher May } IBM Analytical Services } } } } } } }
Clarification: Yes, I am talking about physical micrographs (film/negatives). Sorry about the confusion.
Thanks for all of the suggestions thus far. My first thought was to set up a FileMakerPro database to store the information, but I will look at the commercial packages that have been mentioned.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
At 9:27 AM -0400 7/19/01, Greg Erdos wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ThumbsPlus is good if you have a digital image to keep track of, but } I am not sure if Dennis was referring to that or to physical } micrographs. If it is the latter then I would suggest using } Microsoft Access and setting up their own forms and reports to suit } their particular needs. } Greg Erdos }
{x-charset windows-1252} } I have a JEOL 6320 FEGSEM equipped with a Oxford CT1500 cryo unit which I } have used for a couple of years. I recently replaced the Cr foil/al back } plate target with a Cr sputtered on aluminum target. Now I get a plasma but } no deposition of Cr. Yesterday, I replaced a leaky argon supply valve and } metering valve. Still no deposition. Anyone have any ideas as to what the } problem is? } } I am also looking for access to a FEGSEM with a CT1500 or CT2500 cryo unit } equipped with a Cr target to get some high priority work done. (Preferably } somewhere here in the mid-west). Please contact me with the appropriate } information. } Thanks, } } Dave Williams } dmwilliams-at-dow.com {/x-charset}
I am a graduate student at the university of Texas Health Science Center. As part of a project I wish to perform a spectral analysis of a certain medication. The medication is a crystalline powder in the form of small (approx. 1mm diameter) spheres. I want to section the spheres to 10 to 30 microns so as to obtain a spectral analysis of the entire bead from its outer coating to its interior. I have tried OCT embedding and cyrostat sectioning. However, the beads pulverized during sectioning. Can anyone recommend a way to section these spheres so as to preserve the material as much as possible? I should also point out that the material is freely soluble in ethanol and methanol, as well as water.
Thank you in advance for your help and suggestions.
Below is the result of your feedback form. It was submitted by (dkray-at-siu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, July 19, 2001 at 14:53:44 ---------------------------------------------------------------------------
Email: dkray-at-siu.edu Name: Dave Krayesky
Organization: SIUC
Education: Graduate College
Location: Carbondale, IL 62901
Question: What is a flat field objective and how does it differ from other types of objectives one might use on a microscope?
Can anyone comment on the retention of copper (in dosage experiments) by "staining" with rubeanic acid? What is the complex seen in light microscopy? Would it be accessible to x-ray microanalysis? ......... after removing a coverslip!
Keith Ryan Marine Biological Association Plymouth UK
Following my posting regarding copper/rubeanic acid, does anyone have information about the "pyroantimonate" method. A visitor to my lab has used it (at my suggestion) in an attempt to retain copper in accumulation/stress experiments from a home-base where cryomethods were not possible.
After a week of looking, we see very little in thin/semi-thin sections with XRMA. It is a method where chemical fixation is combined with potassium pyroantimonate to complex and retain the copper (and any other cations?.
Keith Ryan Marine Biological Association Plymouth, UK
Steve Chapman (Protrain Ltd) will be in Australia this October to give courses at the Electron Microscopy Unit of the Australian National University in Canberra. There will be a total of 8 days covering SEM and TEM, divided into 4 modules -you may enrol in one or more modules. Courses will be taught by Steve with assistance from ANU EMU staff.
Brief description below: details and enrolment on www.anu.edu.au/EMU/workshops/01chapman.html
I. SEM MASTERCLASS Tuesday 16th to Thursday 18th October. Three days: AUS$600
II. Maintaining and Monitoring the SEM. Friday 19th October. Pre-requisite: completion of this or a recent SEM Masterclass, or equivalent. One day: AUS $250
III. TEM Maintenance . Monday 22nd October, Tuesday 23rd October. Pre-requisite: familarity with TEM operation. Two days: AUS $500
IV. TEM Operation. Wednesday 24th October, Thursday 25th October. Two days: AUS $400
Meals and accommodation not included.
See you there - your opportunity to learn about EM instrumentation in a very practical way, and have a great time. This will be the third time we have run Module I - so far the recedivist rate is about 30%.
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
Cogent comment. If the coverglass was mounted with the minimum of DPX, then the time required will be "over the weekend" or more. This from long experience with removing coverglasses cracked by students and a common excuse, "I thought it was parfocal!" Cogent comment? If the lab is cold/cool, put it all in a screw cap Coplin jar and set IT, not immersed, in a tepid water bath.
Question. I've never used DPX. Is it's solvent xylene? If it's plastic and it dries to plastic, then you might have to use acetone to get by the perimeter seal.
Irrational comment. OK, I won't remove all the chemicals from the lab.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Keith Ryan } Reply To: kpr-at-mba.ac.uk } Sent: Thursday, July 19, 2001 9:37 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Removing DPX/cover glass } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Greetings from Plymouth, UK } } Can anyone advise on removing a coverslip mounted using DPX? I am } currently soaking it in xylene, but overnight didn't "cut the mustard"! } Any cogent comments? Please?! } } Keith Ryan } Marine Biological Association } Plymouth } } }
We are close to choosing a new SEM and have come to the conclusion that if we want to view salt crystals with liquid inclusions we must do so under positive chamber pressure in order to reduce explosive cleanups. I have not dealt with such specimens before, so I am relying on common sense when expressing this conclusion. The problem is that my conclusion appears to reduce the market to one. I would like to know if I have this right or not.
Thanks,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
Hi All, We have a BioRad/Polaron E-7000 cryo unit (that was attached to a Philips SEM) available to a good home. Please contact me offline. Thanks john shields Univ. of Athens, GA USA jshields-at-cb.uga.edu (706)542-4080
I can't say that I have ever looked at such crystals. Are you hoping to image the inclusions? I know we can see some subsurface voids at higher voltages (20-25kV), but I think you would simply not see much.
How much pressure difference does it take to rupture the crystals? How much internal pressure of the fluid inclusions? Remember that even under absolute vacuum you have only raised the pressure differential by 15 psi. I would think most materials could contain that. Even a 10 torr atmosphere would not spare you much stress. I am not sure what you mean by a scope with positive pressure.
I suppose that there may always be a few inclusions that would blow under vacuum. I would think a pre-pump in a less expensive enclosure would cause those to fail and thus spare the ESEM from harm.
Warren
At 08:52 AM 7/20/2001 -0400, you wrote: } Good Morning Listers, } } We are close to choosing a new SEM and have come to the conclusion } that if we want to view salt crystals with liquid inclusions we must do so } under positive chamber pressure in order to reduce explosive cleanups. } I have not dealt with such specimens before, so I am relying on } common sense when expressing this conclusion. The problem is that my } conclusion appears to reduce the market to one. } I would like to know if I have this right or not. } Thanks, } } Fred Monson } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI)
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I'm assuming (though you don't actually say) that it is thin sections that you are after. If so, I'm not surprised that embedding and sectioning was unsuccessful. You have one of those challenging specimens that profs seem to love handing to grad students, either with the expectation that specimen prep is trivial, or, knowing it is a beast, seeing if the grad student can come up with a new wrinkle!
You don't actually say that it is absolutely necessary for the analysis that the section hold together. Could you do a thin section, say, 100-200nm, do an area analysis of the (dry-sectioned) pulverized bits (lying on a coated grid on the diamond near the knife edge), then trim very coarse for, say 50 microns, then thin section again, and so on? If there is a 30 degree diamond knife around, you might reduce the 'pulverizing' action.
Alternatively, find someone in the neighbourhood with a focused ion beam (FIB) system, usually in Physics or Engineering. We have access to one and have sectioned (though for SEM analysis so we didn't try thin sectioning) some pretty delicate hydroxide powders with no mechanical damage whatsoever. If your particle is 'reinforced' with resin, a FIB-cut section should still be intact after it is cut through and flops on its side, since the incident Ga ion beam is at near zero degrees. The tricky part will come in getting the section onto a grid with something called a micromanipulator (a very fine glass fibre - electrostatic attraction makes the section 'fly up' onto the tip, hopefully intact, then you move the section over the grid and gently press it down). Most FIB types I know would be intrigued by the demanding nature of your specimen.
Good luck!
Tom Malis malis-at-nrcan.gc.ca
} From: Margaret Miller {MILLERMM-at-uthscsa.edu} } Date: Thu, 19 Jul 2001 22:18:21 -0500 } To: Microscopy-at-sparc5.microscopy.com } Subject: Sectioning a crystalline powder sphere } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } -- } To Listers, } } I am a graduate student at the university of Texas Health Science Center. } As part of a project I wish to perform a spectral analysis of a } certain medication. The medication is a crystalline powder in the } form of small (approx. 1mm diameter) spheres. I want to section the } spheres to 10 to 30 microns so as to obtain a spectral analysis of } the entire bead from its outer coating to its interior. I have tried } OCT embedding and cyrostat sectioning. However, the beads pulverized } during sectioning. Can anyone recommend a way to section these } spheres so as to preserve the material as much as possible? I should } also point out that the material is freely soluble in ethanol and } methanol, as well as water. } } Thank you in advance for your help and suggestions.
Dear all, I have found out the source of my particular problem with importing files into DTSA. It was caused by the way I transferred them over the network to the Mac with Fetch. It did not recognise the files as text files and therefore did not treat them as such. When I explicitly told Fetch that they were Text files, however, they could be opened in DTSA.
So, my problem was not caused by different variations in EMSA file formats, but by the fact that the Mac didn't know that they were text files.
Thanks again for all the suggestions, I seem to have set a huge discussion raging by a seemingly innocent comment about difficulties importing my files!
Best wishes to all.
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
} We have inherited a Leitz Metallux 3 microscope with epifluorescence, } however, without a user's manual (not a complaint, just a fact) and are } wondering whether anyone knows a source for old microscope manuals.
Judy -
There's a microscopy book dealer in the UK; try them: http://www.savonabooks.free-online.co.uk
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Being an SEM guy, I wonder about embedding it and dry sanding it to a cross section with SiC paper. Water is normally used for a lubricant, but you could forego that and just use a few more sheets of paper.
Such a sample would not be good for TEM, but I would think an SEM could handle it well enough.
Warren
At 10:18 PM 7/19/2001 -0500, you wrote: } -- } To Listers, } } I am a graduate student at the university of Texas Health Science Center. } As part of a project I wish to perform a spectral analysis of a certain } medication. The medication is a crystalline powder in the form of small } (approx. 1mm diameter) spheres. I want to section the spheres to 10 to 30 } microns so as to obtain a spectral analysis of the entire bead from its } outer coating to its interior. I have tried OCT embedding and cyrostat } sectioning. However, the beads pulverized during sectioning. Can anyone } recommend a way to section these spheres so as to preserve the material as } much as possible? I should also point out that the material is freely } soluble in ethanol and methanol, as well as water. } } Thank you in advance for your help and suggestions.
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Marbaret: I would be tempted to use a metallographic approach (being a metallographer). Go over to the Materials Science guys for guidance in making an epoxy mount. Talk to Dr. Mark Cavaleri of MMM (mecavaleri-at-mmm.com) about polishing methods. He did a complete section of an M&M without losing any of the sugars.
Good luck,
Sam Purdy Teech. Center National Steel Corp. Trenton MI 48183 spurdy-at-nationalsteel.com
Below is the result of your feedback form. It was submitted by (EvanPaassen) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, July 20, 2001 at 14:32:10 ---------------------------------------------------------------------------
Email: EvanPaassen Name: Ed van Paassen
Organization: WAU
Education: Graduate College
Location: City, State, Country
Question: Does anyone know where I can get a used binocular stereo microscope?
Dennis: Altho' I have to use Access from time to time, it is not the program with which I am most familiar. Several years ago (over 10, believe it or not) I used Paradox to develop a database for the lab work going on--both EM and LM. It has been invaluable, and I have been able to maintain things through upgrades (altho' I think I may have missed the next to last one!). There has never been a problem transferring my data from one version to the next. I personally find Paradox easier to work with than Access, but then I started out using WordPerfect, and I still prefer that program. Another (and, _Free_) program is the 602ProSuite of products. I have only used the wordprocessing package so far, but it seems to have most of what I need/use. The program has received great reviews on the ZDNet website. And it's a fantastic price. Back to Paradox--one of the things I like best is the ability to go back and modify your database without losing (or just as bad, having to reenter everything) data. As projects/experiments have expanded/changed or been linked to other studies (via technique, reagents, etc) new categories can be added. Likewise, the search functions permit cross-database searches for seemingly (or unsuspected) relationships.
Roger
No financial interest--just a very satisfied, long-term user. On Thu, 19 Jul 2001 14:37:41 -0400, Dennis C. Winkler wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Greg and others, | | Clarification: Yes, I am talking about physical micrographs | (film/negatives). Sorry about the confusion. | | Thanks for all of the suggestions thus far. My first thought was to | set up a FileMakerPro database to store the information, but I will | look at the commercial packages that have been mentioned. | | - Dennis | | ------------------------------------------------------------------------ | Dennis C. Winkler, PhD. Phone: (301) 496-0131 | Laboratory of Structural Biology Research Fax: (301) 480-7629 | NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov | Bldg. 6, Room B2-26, MSC-2717 | Bethesda, MD 20892-2717, U.S.A. | | | At 9:27 AM -0400 7/19/01, Greg Erdos wrote: | } ------------------------------------------------------------------------ | } The Microscopy ListServer -- Sponsor: The Microscopy Society of | } America To Subscribe/Unsubscribe -- Send Email to | } ListServer-at-MSA.Microscopy.Com | } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | } -----------------------------------------------------------------------. | } | } | } ThumbsPlus is good if you have a digital image to keep track of, but | } I am not sure if Dennis was referring to that or to physical | } micrographs. If it is the latter then I would suggest using | } Microsoft Access and setting up their own forms and reports to suit | } their particular needs. | } Greg Erdos | } |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
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To all, The annual meetings are quickly approaching. We will again hold a session with discussion centering around topics pertaining to Core Facility Management. The topics and session facilitators are listed at the end of this message. Using the same format as last year, the facilitators will give introductions to the topics and then there will be extensive discussion involving all attending.
The session is scheduled for 8:00AM Monday August 5 in Room 201A. I hope to see lots of early risers as, due to later scheduling which partially conflicts with this session, we will have to start on time and move right along to cover the topics.
Please feel free to bring handouts or overhead if you feel they might be useful in the discussions. Course outlines might be very helpful for the first topic. Illustrations to help clarify questions pertaining to scientific ethics in the performance of our duties might also be helpful for the last topic.
Looking forward to seeing old friends and meeting new fellow microscopists at the meeting. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907 ============================================================================
This session is intended to highlight topics that are of concern or interest to managers of microscopy facilities in both academic and industrial settings and with emphasis in life sciences and/or materials areas. There will be three topics discussed at M&M 2001. Each topic will be introduced by one or more facilitators. The introduction will be followed by an extended open discussion with the audience.
Proceedings will be recorded and transcribed so they can be made available to interested persons after the meeting.
1) Training Users: Courses? Workshops? … what works for whom?
Facilitator: John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) Southern Illinois University
2) Calibration of Electron Microscopes: how to do this, how often, pit-falls and problems.
Facilitators: SEM: Michael Postek, Group Leader, Nanometer-scale Metrology National Institute of standards and Technology (NIST)
TEM: John P. McCaffrey National Research Council of Canada
3) Scientific Ethics: what are our responsibilities as facility managers?
Facilitator: Michael Kalichman Director, Research Ethics Program Office of Graduate Studies and Research, 0003 University of California, San Diego
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id RAA22835 for dist-Microscopy; Sun, 22 Jul 2001 17:40:58 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id RAA22832 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 22 Jul 2001 17:40:27 -0500 (CDT) Received: from avocet.mail.pas.earthlink.net (avocet.mail.pas.earthlink.net [207.217.121.50]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id RAA22825 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 22 Jul 2001 17:40:15 -0500 (CDT) Received: from earthlink.net (cpe-24-221-123-240.az.sprintbbd.net [24.221.123.240]) by avocet.mail.pas.earthlink.net (EL-8_9_3_3/8.9.3) with ESMTP id PAA08040 for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 22 Jul 2001 15:37:24 -0700 (PDT) Message-ID: {3B5B5687.3060704-at-earthlink.net}
Listers,
I am looking for an EDS detector mounting flange for a JEOL JSM-IC845 SEM. The detector to be mounted is a Tracor Northern, with a 40 deg take off angle.
If someone has one of these and would like to part with it, please contact me off line. Thanks.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 480.967.3946
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Further clarification.....the date of the session is Monday, August 6 not Monday August 5 as indicated below. Secondly all topics will be covered in order listed below in room 201A. Each topic is assigned 45 min for facilitator's introduction followed by open discussion. I know this may not be sufficient for everyone to speak their piece but it is all the time we were allotted. By keeping on track we can still cover a lot of ground. Perhaps those topics that need more time can be dealt with in more detail at future meetings. Debby
To all, The annual meetings are quickly approaching. We will again hold a session with discussion centering around topics pertaining to Core Facility Management. The topics and session facilitators are listed at the end of this message. Using the same format as last year, the facilitators will give introductions to the topics and then there will be extensive discussion involving all attending.
The session is scheduled for 8:00AM Monday August 5 in Room 201A. I hope to see lots of early risers as, due to later scheduling which partially conflicts with this session, we will have to start on time and move right along to cover the topics.
Please feel free to bring handouts or overhead if you feel they might be useful in the discussions. Course outlines might be very helpful for the first topic. Illustrations to help clarify questions pertaining to scientific ethics in the performance of our duties might also be helpful for the last topic.
Looking forward to seeing old friends and meeting new fellow microscopists at the meeting. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907 ============================================================================
This session is intended to highlight topics that are of concern or interest to managers of microscopy facilities in both academic and industrial settings and with emphasis in life sciences and/or materials areas. There will be three topics discussed at M&M 2001. Each topic will be introduced by one or more facilitators. The introduction will be followed by an extended open discussion with the audience.
Proceedings will be recorded and transcribed so they can be made available to interested persons after the meeting.
1) Training Users: Courses? Workshops? … what works for whom?
Facilitator: John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) Southern Illinois University
2) Calibration of Electron Microscopes: how to do this, how often, pit-falls and problems.
Facilitators: SEM: Michael Postek, Group Leader, Nanometer-scale Metrology National Institute of standards and Technology (NIST)
TEM: John P. McCaffrey National Research Council of Canada
3) Scientific Ethics: what are our responsibilities as facility managers?
Facilitator: Michael Kalichman Director, Research Ethics Program Office of Graduate Studies and Research, 0003 University of California, San Diego
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA24249 for dist-Microscopy; Mon, 23 Jul 2001 08:10:29 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA24242 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 23 Jul 2001 08:09:58 -0500 (CDT) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA24234 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 23 Jul 2001 08:09:47 -0500 (CDT) Received: from sultan (sultan.cb.uga.edu [128.192.63.198]) by cellmate.cb.uga.edu (8.9.3/8.9.3) with ESMTP id JAA11535 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 23 Jul 2001 09:04:28 -0400 To: Microscopy-at-sparc5.microscopy.com
To clarify the previous listing,
The BioRad/Polaron cryo unit was originally attached to a Philips 505 and has been in storage for quite some time. I was told it had problems prior to being boxed up. It will be better suited as spare parts for anyone with an existing unit. Sorry for the previous incompleteness. John Shields EM Lab Univ. of Georgia Athens, GA 30602
I'm not looking to start a flame war here. Please bear with me.
I was going to update my Zeiss Axioplan DIC/phase system to a newer model. I got some major reverberations and feedback that something very bad has happened regarding Zeiss the company. Most feedback seems to be along the lines of bad customer service, no parts, and no support.
My Zeiss systems have been working great for several years and I have not needed any factory support. I ordered some fine clock oil for a condenser and a few other odd and ends and had no problems. My Plan Neofluars don't wear out. So what is the big deal with Zeiss Co. these days?
I have no financial interest in Zeiss. I only have a financial interest in not getting burned. To avoid public acrimony, perhaps private responses off-line are appropriate. I would certainly appreciate some info about what I am obviously missing.
Gary Gaugler, Ph.D. Microtechnics, Inc. 7970 Twin Rocks Rd Granite Bay, CA 95746
Would you confirm us the Core Facility Management session is scheduled for Monday August 6 not Sunday August 5. In case of Aug 5, we have to change our schedule for the flight and hotel.
Thanks
Greg Ning EM Facility Medical College of Wisconsin
Debby Sherman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } To all, } The annual meetings are quickly approaching. We will again hold a session with discussion centering around topics pertaining to Core Facility Management. The topics and session facilitators are listed at the end of this message. Using the same format as last year, the facilitators will give introductions to the topics and then there will be extensive discussion involving all attending. } } The session is scheduled for 8:00AM Monday August 5 in Room 201A. I hope to see lots of early risers as, due to later scheduling which partially conflicts with this session, we will have to start on time and move right along to cover the topics. } } Please feel free to bring handouts or overhead if you feel they might be useful in the discussions. Course outlines might be very helpful for the first topic. Illustrations to help clarify questions pertaining to scientific ethics in the performance of our duties might also be helpful for the last topic. } } Looking forward to seeing old friends and meeting new fellow microscopists at the meeting. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907 } ============================================================================ } } M&M 2001 } Core Facility Management “Experts” session } } This session is intended to highlight topics that are of concern or interest to managers of microscopy facilities in both academic and industrial settings and with emphasis in life sciences and/or materials areas. There will be three topics discussed at M&M 2001. Each topic will be introduced by one or more facilitators. The introduction will be followed by an extended open discussion with the audience. } } Proceedings will be recorded and transcribed so they can be made available to interested persons after the meeting. } } 1) Training Users: Courses? Workshops? … what works for whom? } } Facilitator: John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } Southern Illinois University } } 2) Calibration of Electron Microscopes: how to do this, how often, pit-falls and problems. } } Facilitators: } SEM: Michael Postek, } Group Leader, Nanometer-scale Metrology } National Institute of standards and Technology (NIST) } } TEM: John P. McCaffrey } National Research Council of Canada } } } 3) Scientific Ethics: what are our responsibilities as facility managers? } } Facilitator: Michael Kalichman } Director, Research Ethics Program } Office of Graduate Studies and Research, 0003 } University of California, San Diego } } }
Dear Fred, Please refer to the American Vacuum Society site: http://www.aip.org/avsguide/refguide/pressure.html for a fast and useful education in the meaning of pressure. Through the kindness, and forbearance, of Bill Miller you have been saved from exhibiting your own version of ignorance any further. You should apologize to all the non-vacationing listers for exposing yourself in this manner. In future, some information on the basics would probably save you from this kind of embarrassment. First of all, I suspect you would have been helped if the phrase "variable pressure" had been correctly(?) (mis?)stated as "variable vacuum".[there's that period again!] As Bill said, even the best SEM's cannot operate with the chamber open, and you should, with all of your 'smarts', have been able to see that. Once you have your brain switched to the proper channel, I am certain you will see this point and stop bothering these people. Finally, and I'm sure I speak for all, you should take your own advice and look in the books first. But also, please remember what I always tell my students. The only dumb question is the one you don't ask. Well, as you can see, that's not quite the truth, but it's pretty good advice. And, finally, finally, about your question. Perhaps what you should have asked was how users of SEM technology look at water in the liquid state?
Regards and apologies,
Fred Monson
} ---------- } From: Monson, Frederick C. } Sent: Friday, July 20, 2001 8:52 AM } To: 'Microscopy Listserver' } Subject: SEM, Liquid Inclusions in Salt Crystals } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good Morning Listers, } } We are close to choosing a new SEM and have come to the conclusion } that if we want to view salt crystals with liquid inclusions we must do so } under positive chamber pressure in order to reduce explosive cleanups. } I have not dealt with such specimens before, so I am relying on } common sense when expressing this conclusion. The problem is that my } conclusion appears to reduce the market to one. } I would like to know if I have this right or not. } } Thanks, } } Fred Monson } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI) } Schmucker II Science Center (Room: SS024(Basement)) } South Church Street } West Chester, PA, 19383 } MailDrop: Department of Geology/Astronomy } Phone: 610-738-0437 } Fax: 610-436-3036 } email: fmonson-at-wcupa.edu } Please call before visiting. } }
Evening Keith, Check Wilson's disease. rubeanic acid chelates the Cu, Co or Ni Check Merck Index Ray, Xavier(1961), J Indian Chem Soc, 38:535(review) Have no experiece with EM for this problem. Also: http://www.ninds.nih.gov/health_and_medical/disorders/wilsons_doc.htm
Regards and let us know how the marine bio studies go.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Keith Ryan } Reply To: kpr-at-mba.ac.uk } Sent: Friday, July 20, 2001 3:40 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: copper retention-rubeanic acid } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello Listers } } Can anyone comment on the retention of copper (in dosage experiments) by } "staining" with rubeanic acid? What is the complex seen in light } microscopy? Would it be accessible to x-ray microanalysis? ......... } after removing a coverslip! } } Keith Ryan } Marine Biological Association } Plymouth UK } }
I did set up a Filemaker Pro database years ago to log TEM micrographs as I was shooting them. I used the database to make labels for the micrographs (big time-saver for me), print a list of negatives shot at each session and do annual reviews of project work. It was also useful as a searching tool, for instance if I forgot the date that I last examined samples from a certain project. It would be easy to set up a similar database with a field for micrograph storage information.
Note: the initial plan was to set up the database for a whole microscopy lab of mixed PC and Mac-users, but we had filesharing problems over our network system. I did think Filemaker Pro was easier to use than Access (which I converted to after everything went PC). Don't get me wrong, I like using Access as well. But for creating a simple text-based database Filemaker was very straightforward, even if it had some limitations regarding relational comparisons (it's been so long I can't remember what the limitations were, just that they existed!).
I should emphasize that what made the system most convenient for me was having it set up on the computer attached to the TEM. That way I could enter each record as soon as the negative was exposed, the same way one would make an entry in a logbook. It took a few seconds longer than handwriting, but overall it did not feel like an extra chore to do the data entry.
Caveat: I set this up about 8 years ago, and haven't worked with Filemaker Pro for over 3 years. Things may have changed... But if you are interested I'd be glad to send a copy of the database template (if I can still find it). Hope it works out for you, Karen -------------------------------------------------------------------------------- Karen S. Zaruba BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 kszaruba1-at-mmm.com
"Dennis C. Winkler" {Dennis_Winkler-at-nih.gov} 07/19/01 01:37 PM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: Karen S. Zaruba/US-Corporate/3M/US) Subject: RE: Database of Archived EM Micrographs
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Greg and others,
Clarification: Yes, I am talking about physical micrographs (film/negatives). Sorry about the confusion.
Thanks for all of the suggestions thus far. My first thought was to set up a FileMakerPro database to store the information, but I will look at the commercial packages that have been mentioned.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
To: microscopy-at-msa.microscopy.com,CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Zeiss has always been a company that has supported their products dating back to almost the stone age. It seems that has come to an end. They broke (terminated) their agreements with their whole dealer base, the independent companies who had been supporting Zeiss products for decades. Now they are not supporting microscopes the have just sold a year or two ago. We have a brand new Zeiss microscope in our Emergency Room Lab that has a bad lens. No parts available to repair it!!! I'm supposed to throw away a new $5000 dual head microscope because the company won't support it?
I no longer recommend purchasing Zeiss products.
There have always been other companies who offer much better value then Zeiss. The redeeming value of Zeiss has been in the past that they offered almost unlimited support for their older products. No more.
Dave Burton Optical Specialist
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, July 23, 2001 7:13 AM
Correction - Rather than being included as attachments to the previous message, contents of the Late Breaking Poster Session and the Corporate Members Platform Session have been added to the meeting web site. You can access these by clicking the Symposia icon on the Microscopy and Microanalysis 2001 web site.
------- Forwarded Message Follows ------- Date sent: Mon, 23 Jul 2001 18:22:05 -0500 To: Microscopy-at-sparc5.microscopy.com } From: "Bob Price" {price-at-dcsmserver.med.sc.edu}
Below is the result of your feedback form. It was submitted by (srinivas.mullapudi-at-uth.tmc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 24, 2001 at 10:07:20 ---------------------------------------------------------------------------
We are attempting to visualize pulmonary vasculature. One method would be to perfuse an animal with a fluorescent dye - for example FITC-dextran - but, since we expect to section the lung, the dye would leak out at the severed edges.
Does anyone know of a compound in which to disolve the dye and perfuse the animal with, that would polymerize into a soft solid and remain within the vessels? I remember reading about something like this used to immobilize suspension cells in a culture dish.
Thank you
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
I wish to measure the grain size of 1 micron thick Al0.5%Cu films on Single Crystal Silicon substrate. These are deposited by magnetron sputtering process. Any suggestions or experience out there? Thanks Rick Eller
What kind of results have some of you had with Kodak MDS 100? The price is getting pretty reasonable for the resolution if the software works well enough to be usable. My particular concern it lack of manual control over exposure.
TIA Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
Judy Trogadis wrote: ============================================================== We are attempting to visualize pulmonary vasculature. One method would be to perfuse an animal with a fluorescent dye - for example FITC-dextran - but, since we expect to section the lung, the dye would leak out at the severed edges.
Does anyone know of a compound in which to disolve the dye and perfuse the animal with, that would polymerize into a soft solid and remain within the vessels? I remember reading about something like this used to immobilize suspension cells in a culture dish.
============================================================== I believe the "compound" you are referring to is the Mercox™ Corrosion Casting System, it comes in two different colors (red and green) plus clear (for SEM applications). You can see information about it on our website at URL http://www.2spi.com/catalog/chem/mercox-corrosion-casting.html
You can also see reproduced on the website an article that appeared in Microscopy Today, "Vascular Corrosion Casting Can Provide Quantitative as Well as Morphological Information on the Microvasculature of Organs and Tissues" by Fred E. Hossler, J. H. Quillen College of Medicine, Issue 98-7, Sept. 1998, or see URL http://www.2spi.com/catalog/chem/mercox-casting.html
Disclaimer: SPI Supplies distributes the Mercox kits to researchers worldwide.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
What kind of results have some of you had with Kodak MDS 100? The price is getting pretty reasonable for the resolution if the software works well enough to be usable. My particular concern it lack of manual control over exposure.
Thanks Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
Has anyone any experience with the Nikon Epi-Fluorescence Attachment for a Stereo Microscope. For example how does it compare with the Leica MZ FLIII Fluorescence Stereo Microscope?? Any personal responce will be treated in confidence.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
Would anyone have a copy of a Leitz manual for a Dailux 20 with the epifluorescence head? We have one that we want to use and would like information on the filter sets etc..
The first method that comes to mind, is to ion mill from the Si side, and then look in the TEM. I've done this in pre PIPS days on dimpled samples.
I also recall rigging up a chemical method where the Si was etched away (a mixture of nitric acid and HF. I can't recall the concentrations, but it was likely something standard which others might still be using) leaving the surface film, again for TEM.
If you are handy with a microtome, surface slices coud be taken from a bit of the wafer glued to a dummy block. This will give sections thin enough for TEM, but there will be compression artifacts.
And the last "top of my head" suggestion... at 1 micron thick, are the grains large enough to see using channeling contrast in the SEM?
"Eller, Rick" {ellerro-at-tsmd To: "'Microscopy-at-MSA.Microscopy.Com'" .com} {Microscopy-at-sparc5.microscopy.com} cc: 07/24/01 Subject: Grain Size Measurement of Thin Metal Films ? 04:02 PM
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I wish to measure the grain size of 1 micron thick Al0.5%Cu films on Single Crystal Silicon substrate. These are deposited by magnetron sputtering process. Any suggestions or experience out there? Thanks Rick Eller
Microscopists: We are looking for an environmental chamber that will allow us to heat materials to approximately 1100 C under different reactive gasses while viewing in-situ by either SEM (ESEM) or light microscopy. ESEM would be preferred due to the size range of the subject. We would be interested in contracting this work to your lab, "renting" the chamber or purchasing outright, depending on what's available. Please contact us directly with responses so we don't clutter the list!! Thanks in advance. Bill William A. Heeschen Microscopy, Digital Imaging The Dow Chemical Company 1897 Bldg. Midland, MI 48667-1897 waheeschen-at-dow.com voice: 989-636-4005 fax: 989-638-6443
Actually, one merely has to inject Evans Blue into the experimental animals vasculature, and the albumen in the blood will be instantly bonded by the dye. The dye is supposed to fluoresce only when so bonded. The other way, of course is to set up the albumen-Evans Blue first (see Kodani(?) in '60's for testicular barrier studies)followed by suitable dialysis of unbound EVB (stoichiometry is apparently 1:1). Fixation followed by sectioning with fluorescence. FITC-?, though bound with greater avidity than plain fluorescein does not elicit the dye fastness of EVB with albuminoids.
Evans Blue Dye Site (Blood volume): http://www.evansblue.co.uk/index.html
This is everything but the trip to the confocal (see Czymmek, K (2000), "Imaging and volumetric quantitation of vascular corrosion casts with laser scanning confocal microscopy, Proceedings: Microscopy and Microanalysis 2000, Philadelphia, PA, August 13-17, 2000, p 562. (kirk.czymmek-at-mvs.udel.edu). Using Mercox Blue corrosion cast, excite 633nm laser, ~660nm barrier.
Fred
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Judy Trogadis } Sent: Tuesday, July 24, 2001 3:49 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: compound for immobilization } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow Microscopists : } } We are attempting to visualize pulmonary vasculature. One method would be } to perfuse an animal with a fluorescent dye - for example FITC-dextran - } but, since we expect to section the lung, the dye would leak out at the } severed edges. } } Does anyone know of a compound in which to disolve the dye and perfuse the } animal with, that would polymerize into a soft solid and remain within the } vessels? I remember reading about something like this used to immobilize } suspension cells in a culture dish. } } Thank you } } } } } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital, 8Q } 30 Bond St. } Toronto, ON M5B 1W8 } } ph: 416-864-6060 x6337 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } }
I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results.
Good Afternoon! This is a follow up question involving tissue sections prepared for the TEM. My original question dealt with perfusion with physiological saline followed by cold 4% paraformaldhyde. I will be incubating the tissue (dorsal root ganglions) with an antibody. I have spoken to my advisor and Franklin Bailey at the UI EM center about preparing the tissue sections and eventual incubation with antibodies. I have received conflicting advice and am putting this out for more advice. Since I am going to use antibodies, does this change how I perfuse the animal? When incubating with the antibodies, should I do it pre- or postembedding? Immunogold labeling may be used and I have been told that preembedded labeling may damage the knife. I am open to other ways of labeling this tissue. Thank you.
Meghan Towner Graduate Student Biological Sciences University of Idaho
Try smoking some Mo with heat to produce a white smoke and wave your sample in the smoke. You will pick up small particle of MoO3. Alternatively, you could burn Mg and wave your sample in the smoke. Experiment with how high in the smoke you wave your sample.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Smartech [mailto:smartech-at-optonline.net] Sent: Wednesday, July 25, 2001 2:51 PM To: To all on the list
I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results.
Our company needs to get a microscope for inspection and assembly/disassembly tasks. We would like it to have the following characteristics:
1) Stereo viewing 2) Wide field of view 3) Prefer zoom capabilities, but could take anything that would allow magnifications from about 4X to, say, 20X 4) Good working distance
Would anyone care to make any recommendations or do you have any suggestions for us?
Vendor input is welcome. Contact information follows my sig.
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
SMARTech asked the following: =========================================================== I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results. =========================================================== We faced a similar kind of question involving a Mylar film (some years ago), which also was smooth and featureless at the SEM level and there was the need to look for evidence of etching. In this case, the "etch" was exposure to ozone. Our approach was the following:
a) Apply on one side of the film a special silicone resin; this was an earlier version of the silicone used in our SPI Wet Replica Kit (see URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.html ) The line between the resin covered side and the side without resin is quite distinct. The resin thickness is about 0.5 to 1 mm.
b) Do your exposure of the film, but the silicone layer protects the one side from exposure and it will not be harmed by the acid environment. Of course, at some point it can ( I presume) be effected by acid so it has to be a matter of time, temperature and pH.
c) After exposure, the silicone can be removed, and if indeed there is some etching, if differences are seen one side of the interface vs. the other, then you could draw some conclusions.
In those days, we found we needed ordinary Pt/C surface replication methods (and TEM) to show the differences, one side to the other. But in the end one is faced with having to prove the null hypothesis: If you see no effect , does that mean no etching or does that mean one has not looked close enough. Today, I would expect that AFM would have a better chance at seeing differences that might not be seen with Pt/C surface replication.
Another approach, if we were to be doing this today, would be to try applying the approach that has been used by Japanese researchers at Nagoya University (if I remember correctly) who have been using thin osmium metal layers, plasma coated, that are not more than ~1 nm thick, to replicate macromolecules on flat surfaces. The method does work well for DNA for example and might work well for a surface with a very subtle etch effect. If you would like more information about this method, I could share additional information with you about how this is done. I could also dig out my notes on the people who are doing this work.
Disclaimer: SPI Supplies offers this type of silicone resin and also, distributes the OPC-60 Osmium Plasma Coater made by Nippon Laser and Electronics Labs, Nagoya, Japan.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
ie the anti-backstreaming trap between the mechanical pump and the diff pump:-
is there a consensus out there on what's best?
Molecular seive, activated alumina pellets, or Cu gauze?
I've never been that happy about the first two because of mechanical abrasion releasing abrasive powder into the rotary pump, particularly from those Edwards holders that seem designed to mount directly on the pump, where they get nicely and continuously vibrated.
And how do people regenerate each absorbent?
Heat in air to 600 deg C or so?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
You might consider an Olympus SZ-40 system with a Fostec fiber optic ACE ring light. There are several sources for these 'scopes as used. The Fostec may be only available as new (I have a near new light source I would sell...no ring light bundle). I like the Fostec very much.
On the surplus market, Nikon stereo zooms come up quite often. Maybe one of these would work for you. Try http://dovebid.com for options.
gary g.
Disclaimer: No financial interest in any company referenced above.
At 02:31 PM 7/25/2001, you wrote:
} Our company needs to get a microscope for inspection and } assembly/disassembly tasks. We would like it to have the following } characteristics: } } 1) Stereo viewing } 2) Wide field of view } 3) Prefer zoom capabilities, but could take anything that would allow } magnifications from about 4X to, say, 20X } 4) Good working distance } } Would anyone care to make any recommendations or do you have any suggestions } for us? } } Vendor input is welcome. Contact information follows my sig. } } Bruce Girrell } Microline Technology Corp. } 2397 Traversefield Dr. } Traverse City, MI 49686 } http://www.microlinetc.com } } (231) 935-1585 (Voice) } (231) 922-5099 (FAX) } bigirrell-at-microlinetc.com
Anyone out there who can help me. We want to 'refurbish' an old Leitz DURIMET Microhardness Tester for routine applications by students. However, both objectives are badly damaged and beyond repair. (No wonder, the unit is approximately 50 years old and had been used by numerous students in the past). We would like to 'pick up' or purchase therefore the following types of objectives (used or new): Low Magnification 40x and High Magnification A 0.70 CHM 6.3 40x to suit the DURIMET. Thanks for any advise.
Hans Brinkies Senior Lecturer Swinburne University of Technology School of Engineering and Science Industrial Microscopy Laboratory PO Box 218 - Hawthorn - Victoria - 3122 - Australia Phone: +61 3 9214 8657 - FAX: +61 3 9214 8264
Just my two cents concerning pre-embed labeling with gold damaging the knife. We have done it for years and never noticed any damage.
Bob Underwood Morphology Core U of Washington
On Wed, 25 Jul 2001, Meghan Towner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good Afternoon! } This is a follow up question involving tissue sections prepared for the TEM. My original question dealt with perfusion with physiological saline followed by cold 4% paraformaldhyde. I will be incubating the tissue (dorsal root ganglions) with an antibody. I have spoken to my advisor and Franklin Bailey at the UI EM center about preparing the tissue sections and eventual incubation with antibodies. I have received conflicting advice and am putting this out for more advice. Since I am going to use antibodies, does this change how I perfuse the animal? When incubating with the antibodies, should I do it pre- or postembedding? Immunogold labeling may be used and I have been told that preembedded labeling may damage the knife. I am open to other ways of labeling this tissue. Thank you. } } Meghan Towner } Graduate Student } Biological Sciences } University of Idaho } } }
Dear Smartech, Just off the top of my head: 1. clay, ultra-soniced in alcohol and dropped on to dry. 2. the fine aluminum powder used in makeup, dusted on with a paintbrush or dropped from alcohol as in 1. 3. small latex sheres used to calibrate TEMs. Already in suspension, just drop on to dry. I have used all three for various purposes. I have also used a very fine grid (~1000 mesh) laid tight over the sample and then evaporated gold through it. The resulting gold squares are quite visible in the SEM but avoid the problems of edge effect. At 02:50 PM 7/25/01 -0400, you wrote:
} I want so show that a polymer film has not been etched after exposure to an } HCL based environment. The problem is that it is extremely smooth to begin } with. I would like to add some sub-micro features as a background so that I } am not generating a featureless gray photomicrograph. These features would } be essentially a topographic reference. I though of a mono layer of } particles dusted on the surface. I stated w/ common baking flour. The } particles were too large and not stuck to the surface so well (lots of edge } effect) so that one can't effectively have a good contrast of the spheres } and the background in the same micrograph. A very fine grid would also be } nice, but perhaps expensive. Smaller and flatter particles that were put in } solvent and allowed to dried might do the trick. Anybody have any } interesting suggestions or materials? I would be glad to share the results. } } SMARTech
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
The various types of foreline traps are discussed in some detail on pp. 146 - 149 of my book 'Vacuum Methods in Electron Microscopy" (for a description see http://www.2spi.com/catalog/abooks/book48.html/ and http://www.pup.princeton.edu/titles/6484.html/)
At the time I was writing this book it appeared that perhaps the best method of trapping oil vapors from the roughing pump was to use a Micromaze trap produced by the Lesker company. These traps are provided with a heater to be used for regenerating the trap, and it appeared advantageous to run this heater continuously at about 75% full power. Evidence available then indicated that when hot the ceramic Micromaze material caused the oil molecules to break down into small fragments that could pumped away. See above reference for details.
Maybe some more efficient type of trap has been developed since, but I have not heard of it. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Does anyone remember the sucrose concentrations needed to form a gradient for separating cells and cell fractions from media debris? We once used this technique, but I have lost all of my references on the procedure. Thanks
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
Dear Folks, I'm in the process of setting up a system for doing time-lapse video microscopy and would love to get information from others on what set-ups work best for them. I'm interested in people's experience with time-lapse VCRs and digital capture. I'm recording in color using a three-chip CCD video camera. We have a Scion capture card on a Mac G4. I will be recording over 24-hour periods. In particular, has anyone been using NIH Image or Imagej to do time-lapse in color?
I am new to this listserv, so my apologies if I'm raking over old territory. I did do a search of the listserv archives and didn't find any postings in this subject. Many thanks, Mary =============================================== Mary_Tyler-at-umit.maine.edu or mtyler-at-maine.edu
Mary S. Tyler Professor of Zoology Dept. of Biological Sciences University of Maine Orono, ME 04469-5751 FAX: 207-581-2537
You can smoke Mo in a number of ways. You can use wires or boats across the leads of an evaporator and do it in air, or you can take a wire and heat it with a Bunsen burner, or propane torch. The smoke should not be that hot, just don't keep your sample in the smoke or near the radiant heat for too long. The wire should continue to smoke a little just as you remove the heat. If you collect the smoke from Mg, you should be able to collect it far from the heat source as the smoke rises. I assumed in my original post that you would need high resolution imaging and small particles distributed with no carrier media to put the stuff on the surface. There was a response to my earlier post that said to scratch the surface, but that would produce features too gross.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Smartech [mailto:smartech-at-optonline.net] Sent: Wednesday, July 25, 2001 8:08 PM To: Walck, Scott D.
I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results.
POST-DOCTORAL POSITION in Electron Microscopy of Electroceramic Grain Boundaries
A postdoctoral position is available in the area of electroceramics beginning September 1, 2001. The research project focuses on understanding solute segregation at electroceramic grain boundaries, in particular TiO2 and BaTiO3, and the ramifications for electrical transport properties. Aspects of the project will be conducted under the auspices of the Center for Dielectric Studies at the Pennsylvania State University. Through a variety of electron imaging and spectroscopy techniques we aim to quantify solute segregation and grain boundary impedance as a function of doping level and microstructure. For special boundaries we will obtain atomic structure and site-specific segregation information that will be used as the basis for theoretical calculations. The ideal candidates for this position will have experience in HREM, STEM, EDS and/or EELS and have an understanding of grain boundary phenomena in electroceramics. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to Professor Elizabeth Dickey:
Department of Materials Science and Engineering The Pennsylvania State University 195 Materials Research Institute Building University Park, PA 16802-7003
tel: (814) 865-9067 FAX: (814) 863-8561 email: ecd10-at-psu.edu -- Elizabeth Dickey Department of Materials Science and Engineering Pennsylvania State University 195 Materials Research Institute Building University Park, PA 16802-7003
A colleague has asked if I could help explain why the excitation filter in his DAPI filter set on a Zeiss Axiophot deteriorates after a few months of use.
Excitation filters in other sets last longer and don't show this problem. The symptoms are uneven illumination that shows up in the image and an obvious 'burned ' look to the filter.
His questions are like: Is this normal? If not, what is wrong? How can I avoid this problem?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a couple Micromaze bakeable foreline traps that Lesker sells. They were on a TEM that I used to own. I almost forgot that I had these. I kept them with the intention of trying to sell them someday. These two were modified for safety reasons and so nothing around them would get damaged by the heat from these. These traps when turned on get pretty hot if I remember right. For these reasons longer stainless pipes were welded on and a cage was built around each one. I would recommend something like this if you use these traps.
Not only are the maze traps better, I think, but they are easier to use. Use to simply plug them into an AC outlet overnight once a week to bake them out. Kept the vacuum system running while bakeing.
Joel McClintock EM Specialist U of Kentucky
At 02:01 PM 7/26/01 GMT+1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's better to use PERCOLL solution. It's isoosmotic and you could separate cells from microsomal fraction or viruses etc. You may also separate different cell types by this technique. PERCOLL was manufactured by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey
At 11:06 AM 7/26/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I have a question about using Percoll for TEM - someone told me just last week that you shouldn't use Percoll if you'll be doing thin sections, since it is a silica (can damage knives) - has anyone used it and either seen or not seen gunk in their sections? Scratches &/or damaged a knife? Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't resist)
I'm not planning to use it any time soon, but hearing about it again makes me curious about using it to prep for EM. I've seen nasty sucrose crystals a few times with samples from sucrose gradients, but they are mostly an annoyance.
Thanks!
Tamara
On Thu, 26 Jul 2001, Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } It's better to use PERCOLL solution. It's isoosmotic and you could } separate cells from microsomal fraction or viruses etc. You may also } separate different cell types by this technique. PERCOLL was manufactured } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey } } At 11:06 AM 7/26/01 -0700, you wrote:
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
I agree with the ideas of placing simple structures on the surface of the specimen, Mn smoke or polystyrene latex. However general tips for featureless specimens are -
1. Keep the kV down ( {5kV) - to restrict the probe to a true surface investigation
2. Use high tilt (} 45deg) as the small amount of backscatter present will serve to emphasize topography.
Happy scanning
Steve Chapman Senior Consultant Protrain For professional training in electron microscopy world wide www.emcourses.com
Some time ago we tried to section some epoxy-embedded, Percoll-separated cell fractions. This resulted in very evident damage to glass knife cutting edges - to such an extent that we decided not to jeopardize a diamond knife.
Separating the fractions on a sucrose gradient resulted in blocks that sectioned without any problems.
Jan Coetzee
Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Tamara Howard wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
School of Mechanical, Materials, Manufacturing Engineering & Management Advanced Materials Research Group University of Nottingham, UK
Professor of Materials
Applications are invited for the above post from candidates with an outstanding record of research to reinforce and add to existing research strengths and providing a third professorial post in the Advanced Materials Group. Candidates should have expertise in one of the following areas: electron microscopy applied to the processing of advanced materials; surface engineering of materials; near-net-shape processing of materials or biomaterials
The person appointed will be expected to contribute to the teaching of courses within the School, which was graded 4A in the Research Assessment Exercise and gained excellent scores for its teaching quality.
Salary will be within the professional range, minimum £41,089 per annum.
Informal enquiries may be addressed to Professor D G McCartney or Professor A B Seddon, tel: 0115 951 3740, Email: Angela.Seddon-at-Nottingham.ac.uk Further details of the School are available on the WWW at: http://www.nottingham.ac.uk/school4m/.
Further details and application forms are available on the WWW at: http://www.nottingham.ac.uk/personnel/ or from the Personnel Office, Highfield House, The University of Nottingham, University Park, Nottingham, NG7 2RD. Tel: 0115 951 3266. Fax: 0115 951 5215. Email: June.Pedlar-at-Nottingham.ac.uk. Please quote ref. MCM/165. Closing date: 15 September 2001.
…………………………………………………………………………………………………………
To be published as follows:
Guardian - Tuesday 24 July THES - Friday 27 July Jobs.ac.uk - w/c Monday 23 July
Back in the early to mid 80's, I used Ficoll-Hypaque to layer out different white cells from peripheral blood. Never had a problem with sugar crystals. Only damage to the diamond knife was (cringe) user error.
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For what its worth:
Some time ago we tried to section some epoxy-embedded, Percoll-separated cell fractions. This resulted in very evident damage to glass knife cutting edges - to such an extent that we decided not to jeopardize a diamond knife.
Separating the fractions on a sucrose gradient resulted in blocks that sectioned without any problems.
Jan Coetzee
Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Tamara Howard wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Serge y } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id LAA11998 for dist-Microscopy; Fri, 27 Jul 2001 11:36:53 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id LAA11994 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 27 Jul 2001 11:36:23 -0500 (CDT) Received: from thor.oit.pdx.edu (thor.oit.pdx.edu [131.252.120.40]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id LAA11987 for {Microscopy-at-sparc5.Microscopy.Com} ; Fri, 27 Jul 2001 11:36:08 -0500 (CDT) Received: from medusa.oit.pdx.edu (medusa.oit.pdx.edu [131.252.120.41]) by thor.oit.pdx.edu (8.11.1/8.11.1) with ESMTP id f6RGXHU14503 for {Microscopy-at-sparc5.Microscopy.Com} ; Fri, 27 Jul 2001 09:33:17 -0700 (PDT) Received: (from sec-at-localhost) by medusa.oit.pdx.edu (8.11.1/8.11.1) id f6RGXGo23355 for Microscopy-at-MSA.Microscopy.Com; Fri, 27 Jul 2001 09:33:16 -0700 (PDT) X-Authentication-Warning: medusa.oit.pdx.edu: sec set sender to lifengd-at-pdx.edu using -f To: Microscopy-at-sparc5.microscopy.com
Roommate wanted:
I have made a reservation at Hotel Queen Mary in Long Beach, California for the
Microscopy and Microanalysis 2001 Conference from August 5 – 9. It turns out the room has two double beds. If any female colleagues (one female) who want to
share the room with me, please contact me by e-mail (jiaoj-at-pdx.edu). The room is reserved for 4 nights ($115/night for two persons) from August 5 – August 9.
Dr. Jun Jiao Assistant Professor Physics Department Portland State University Portland, Oregon, U.S.A.
I have used Percoll for leydig cell seperations and TEM with no knife damage. After the isolation procedure I have used a series of buffer washes before dehydration and embedding procedures.
Al Kruger Reproduction Research Unit USDA-ARS-MARC Clay Center, NE 68901
Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Mary Tyler } Sent: Thursday, July 26, 2001 3:11 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM time-lapse video microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Folks, } I'm in the process of setting up a system for doing time-lapse video } microscopy and would love to get information from others on what set-ups } work best for them. I'm interested in people's experience with time-lapse } VCRs and digital capture. I'm recording in color using a three-chip CCD } video camera. We have a Scion capture card on a Mac G4. I will be } recording over 24-hour periods. } In particular, has anyone been using NIH Image or Imagej to do time-lapse } in color? } } I am new to this listserv, so my apologies if I'm raking over old } territory. I did do a search of the listserv archives and didn't find any } postings in this subject. Many thanks, Mary } =============================================== } Mary_Tyler-at-umit.maine.edu or mtyler-at-maine.edu } } Mary S. Tyler } Professor of Zoology } Dept. of Biological Sciences } University of Maine } Orono, ME 04469-5751 } FAX: 207-581-2537 } } }
In the old days, filters could be 'burned' out by the high voltage Hg lamp if there wasn't a proper buffer. Is the bulb correct for the illuminator? Sometimes these days, a neutral density filter is placed between the illuminator and the filter set.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Thursday, July 26, 2001 5:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM: DAPI excitation filter damage? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } } A colleague has asked if I could help explain why the excitation filter in } his DAPI filter set on a Zeiss Axiophot deteriorates after a few months } of } use. } } Excitation filters in other sets last longer and don't show this problem. } The symptoms are uneven illumination that shows up in the image and an } obvious 'burned ' look to the filter. } } His questions are like: Is this normal? If not, what is wrong? How can I } avoid this problem? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
First I would like to thank everyone for their help and suggestions = with my problem of "dirty grids" or as I call it (technical term) ka ka = on the sections.
I know it has been a while since I asked this question, but I have not = been able to go through and try out all the suggestions sent to me .
The solvent I am using is a mixture of 50% Methanol and 50% ddH20 Staining was done on dental wax for 10 minutes.
For lead staining it was done for 5 minutes with sodium hydroxide = pellets in a covered petri dish.
Everything was filtered through .45 micron filters.
Solutions I was given for my staining artifact were:
1. Change filter size to .20 microns. 2. Use parafilm instead of dental wax for staining. 3. Shorten my staining time from 10 minutes to 5 minutes. 4. Change suppliers of UA stain 5. Try a new batch of stain.
Solutions I tried.
1. Used parafilm instead of Dental wax 2. used .20 micron filters 3. mixed up a new batch of stain.
By changing all these criteria I fixed the staining problem that occurred a few weeks ago.
Now I get about 90-95% clean grids. Sometimes there is a little schmutz around, but not like occurred before.
Thanks to everyone for their suggestions
Eric A. Rosen UCLA Medical Center Electron Microscopy Lab Dept. Pathology and Lab Medicine Los Angeles, CA 90095 310-825-5737
I haven't had trouble with clays but certainly with gypsum (CaSO4 2H2O). I imagine a very hydrous clay would cause similar problems achieving vacuum. You could dehydrate the sample in a (vacuum) oven, but then of course you're looking at a different clay from the one you started with. Some variety of Environmental or Cryo-SEM might be required, depending on your needs.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Friday, July 27, 2001 8:03 AM, Alun [SMTP:alun-at-deepsouth.co.nz] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Has anyone had any experience with SEM photographing minerals, } or had a problem in obtaining a vacuum in the case of clay type minerals. } } Bernard. } }
I don't think that filtering achieves much since that will only eliminate the boulders; 0.5um rocks are 5mm at only 10,000x mag. Use 2x distilled water.
UA in methanol goes off much faster than UA in water and that may be your major problem. Made up in water I trust UA for one month, when refrigerated and mostly kept in the dark. Made up in methanol, I would trust that solution only for a day or two.
The second problem is over-staining. Ten or even twenty minutes for aqueous UA is normal, but methanolic UA may be over-staining after 2 minutes or so.
I like your technical term "schmutz" - for the uninitiated, schmutzig is German for "dirty" and schmutz means dirt or just muck. I vote that "schmutz" should be official jargon in microscopy. Another little hint: We all have noticed that a grid when applied to a drop of staining solution floats to the top. So does all the schmutz. It makes sense to apply a grid to the side of a drop, that way any top floating schmutz is not picked up but pushed away. With aqueous UA and lead stain it makes sense to only stain one side of the section. Methanolic stains may not support the grid and the grid must be placed into the drop. This increased the chance of contamination and also causes more intense staining. More intense staining may sound desirable, but there are some negatives. This contributes to over-staining, also, some stains only penetrate part of a section and so give the effect of a thinner section and sharper image. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, July 28, 2001 4:28 AM, Eric [SMTP:biology-at-ucla.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the Microscopy List, } } } First I would like to thank everyone for their help and suggestions = } with my problem of "dirty grids" or as I call it (technical term) ka ka = } on the sections. } } I know it has been a while since I asked this question, but I have not = } been able to go through and try out all the suggestions sent to me . } } } The solvent I am using is a mixture of 50% Methanol and 50% ddH20 } Staining was done on dental wax for 10 minutes. } } For lead staining it was done for 5 minutes with sodium hydroxide = } pellets in a covered petri dish. } } Everything was filtered through .45 micron filters. } } } Solutions I was given for my staining artifact were: } } 1. Change filter size to .20 microns. } 2. Use parafilm instead of dental wax for staining. } 3. Shorten my staining time from 10 minutes to 5 minutes. } 4. Change suppliers of UA stain } 5. Try a new batch of stain. } } } Solutions I tried. } } 1. Used parafilm instead of Dental wax } 2. used .20 micron filters } 3. mixed up a new batch of stain. } } By changing all these criteria I fixed the staining problem that } occurred a few weeks ago. } } Now I get about 90-95% clean grids. Sometimes there is a little } schmutz around, but not like occurred before. } } Thanks to everyone for their suggestions } } Eric A. Rosen } UCLA Medical Center } Electron Microscopy Lab } Dept. Pathology and Lab Medicine } Los Angeles, CA 90095 } 310-825-5737 } }
I know that this server and most universities and other large institutions are virus protected, but many readers will be interested to learn that my usual tally of (eliminated) virus messages is about 2 per week. During the last ten days this has blown out 26. My server scans for and eliminates the virus part of a message and we have our own anti virus system. Just now, a lot of people out there are badly affected by the TROJ_SIRCAM.A virus and its variants. All of these message come in with different headers and all these files terminate with these extensions: exe, com, pif, bat, 1nk, scr. I received a couple of virus messages from SE Asian Universities.
Incidentally: That amazing Nigerian scam was supposedly squashed. Hmmm, I used to get one or two messages weekly, now I get ten or more from them and some clones. Trouble is that emails are too cheap to send and since no legislator is tough on spaming, we all must waste our time (and some fools part with their money). Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
I was wondering if anyone could offer some advice regarding my 2010F gun.
I periodically measure beam current using the small viewing screen after aligning the microscope in a reproducible configuration (A1 and A2 set at specific values, 50micron condenser aperture, CBD mode 1nm alpha 9, 1MX magnification with beam focused using Brightness knob). Over the last several months I have observed that the size of the spot is approximately 4 times larger than when the microscope was commissioned in early 1999. The FWHM of the probe was measured yesterday to be 2.2nm. The emission timer reads 19,200 hrs.
Obviously our analytical capabilities are compromised by the gun's current condition but we can still perform the majority of our work.
Is the probe size likely to continue to get larger?
Is our emitter about to expire completely?
What are the chances of the gun lasting another 6 to 12 months without the probe deteriorating dramatically?
Thanks for any advice you can offer, cheers
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
We are seeking to buy an operational 100CX objective lens power supply. Please contact me off line at the address below if you have one for sale.
Thanks in advance.
Owen
Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Department of Materials Science and Engineering Rm 512 M&M Building Houghton, Mi 49931 Ph: 906-487-2002 FAX: 906-487-2934 Email: opmills-at-mtu.edu URL: http://www.mm.mtu.edu/~opmills/index.html
Hello fellow microscopists, has anyone out there know how I can get the free trial version ACDSee 16 bit installation software suitable for Win 3.11 operating system? I tried getting it from their web site but it appears they have discontinued it -only programs suitable for Win 95 and higher.I know that until recently this free down loadable version was available on their website. I have tried contacting their helpline, but so far to no avail. I'd be very grateful to anyone who could contact me off-line with information where I can get it. Or perhaps does anyone know of something very similar -allowing very quick viewing of images in file manager of 3.11? Regards Martin Roe
Hello fellow microscopists, has anyone out there know how I can get the free trial version ACDSee 16 bit installation software suitable for Win 3.11 operating system? I tried getting it from their web site but it appears they have discontinued it -only programs suitable for Win 95 and higher.I know that until recently this free down loadable version was available on their website. I have tried contacting their helpline, but so far to no avail. I'd be very grateful to anyone who could contact me off-line with information where I can get it. Or perhaps does anyone know of something very similar -allowing very quick viewing of images in file manager of 3.11? Regards Martin Roe
I read your message just after carrying out my monthly emitter checks on our 3000F at 20,386 hours. We also hope to predict the degradation in emitter performance and plan a service replacement for a few minutes before the tip fails:-) Well the're not cheap are they?
I have a slightly different set up; I adjust A1 to get 200uA emission and record similar data to you. I also record the beam current on a picoammeter. I don't measure the spot size accurately but I note the magnification required to fill the centre spot on the small screen. It's a subjective test but the result has been either 1, 1.2 or 1.5M times at every test and the spot does not show any trend towards getting larger.
What I have noticed is that the spot size is very dependent on the objective lens setting (now standardised) and beam tilt (rotation centre). If there is any objective astigmatism then it is 'corrected' by the condenser stigmators giving a larger spot.
If you do measure the current on the small screen you may find that it alters with both probe size and position on the screen (different absorption at different positions).
Make sure you are at DV=0 (or -14 in our case to allow for the GIF voltage offset) and that the column is always fully aligned in the mode you use (in our case CBD, alpha 9, spot 0.4nm and smallest C aperture).
I know that Sheffield University were monitoring their 2010F emitter but I don't think that they got any warning of a tip failure before their's was replaced.
Maybe others have better information for you.
Regards, Ron
On Mon, 30 Jul 2001 17:19:26 +1000 Mark G BLACKFORD {mgb-at-ansto.gov.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I was wondering if anyone could offer some advice regarding my 2010F gun. } } I periodically measure beam current using the small viewing screen after } aligning the microscope in a reproducible configuration (A1 and A2 set at } specific values, 50micron condenser aperture, CBD mode 1nm alpha 9, 1MX } magnification with beam focused using Brightness knob). Over the last } several months I have observed that the size of the spot is approximately 4 } times larger than when the microscope was commissioned in early 1999. The } FWHM of the probe was measured yesterday to be 2.2nm. The emission timer } reads 19,200 hrs. } } Obviously our analytical capabilities are compromised by the gun's current } condition but we can still perform the majority of our work. } } Is the probe size likely to continue to get larger? } } Is our emitter about to expire completely? } } What are the chances of the gun lasting another 6 to 12 months without the } probe deteriorating dramatically? } } Thanks for any advice you can offer, cheers } } } Mark Blackford } Materials Division, ANSTO } PMB 1, } Menai, N.S.W., 2234 } Australia } } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed. }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I am considering options for a CCD on a 35mm port or in the PEELS compatible location of a Gatan PEELS. Any input, including from vendors, would be useful. This would be for HREM applications.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Are you aware that in some german dialects "Schmutz" does means other things that dirt? And accidentally some people have family names as Schmutz and even worst they happen to be quite good microscopists using this server as well. May be some latin name or older "dead" language name would help? Or why not leva is as dirt? Just a suggestion. svetla -- Svetla Stoilova-McPhie, PhD The Burnham Institute, 10901 N. Torrey Pines Road, La Jolla, CA 92037 tel: 858 646 3100 ext.3484, fax: 858 646 3196
Hello everyone, All of my previous experience in preparing thick and thin sections for LM and TEM, respectively, is with epon. Should I expect similar behavior with araldite when sectioning with a diamond knife and staining with toluidine blue or aqueous UA and Reynolds lead citrate? I would appreciate any tips or references. Thanks.
Donald Gantz Dept. Physiology & Biophysics Boston University School of Medicine Boston, Massachusetts 02118 Email: Gantz-at-Biophysics.bumc.bu.edu Phone: 617-638-4017
The Microscopy Society of America is seeking an Archivist to manage the historical documents of our Society. This position is appointed by MSA Council for a period of three years.
We are looking for someone who is experienced in the storage and cataloging of (primarily) paper documents. Although this is a voluntary position (non-paying), the Society would provide funding for expendibles and day-to-day operations (and possibly a part-time, student-worker assistant).
If interested (or if you know of a qualified person), please contact me at the M&M 2001 meeting in Long Beach next week or at the following address:
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I have called Hitachi about information on the pumps, but they are having some difficulty in locating any specs on them. Searching the Internet did not give any information either.
The pumps plug into what appears to be a standard 3-prong socket and the pumps have information on the housing for 110 V, so I'm thinking that the power requirements are fairly standard. What I don't know is what flow rate, et cetera, to get for the replacement pumps.
Does anyone know where I could locate the pump specs or could you recommend some pumps that would work? And, in case you haven't figured it out already, I'm not an expert on vacuum systems, so I may be going about this the wrong way by looking at specs that really don't matter. If someone could let me know what parameters are the important things to look for, I'd be grateful.
Postdoctoral Research Associate in Quantitative Electron Microscopy of Nano-Structures
Applications are invited for a postdoctoral research associate position at the Dept. of Materials Science and Engineering, University of Illinois-Urbana/Champaign. The position involves developing electron diffraction and spectroscopy techniques for quantitative characterization of the electronic and structural properties of nano-structures, using the state of art electron microscopy facilities (JEOL 2010F-GIF and VG HB501-STEM) at the Center for Microanalysis of Materials. The successful candidate should have strong experience in advanced materials electron microscopy and diffraction techniques. Experience with STEM, energy-filtered imaging and diffraction, and electron energy-loss spectroscopy is desired, but not required. Preference will be given to candidates with strong experimental aptitude. A PhD. in Physics, Materials Science or related field is required. The position is funded by DOE and is initially for one year with possibility for renewal up to 3.5 years. Applicants should send a curriculum vitae, including a list of publications and research experience, and the names of three references to: Prof. J.M. Zuo, Dept. of Materials Science and Engineering, University of Illinois at Urbana and Champaign, 1304 W Green Street, Urbana, IL 61801; 217-333-2736 (fax); (217) 244-6504 (phone); jianzuo-at-uiuc.edu (email).
Application materials should be received before Oct. 1, 2001 for full consideration.
Dear Mary, As you've been told, NIH Image will do this, saving the timelapse images as a stack held in RAM. However, Object Image, the extended version of NIH Image (avalilable from simon.bio.uva.nl), will do it with a means to create a similar control window to add integration or averaging, and to save the image series to hard drive. I will send you the macro for Object Image directly.
Regards, Glen
Mary Tyler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Folks, } I'm in the process of setting up a system for doing time-lapse video } microscopy and would love to get information from others on what set-ups } work best for them. I'm interested in people's experience with time-lapse } VCRs and digital capture. I'm recording in color using a three-chip CCD } video camera. We have a Scion capture card on a Mac G4. I will be } recording over 24-hour periods. } In particular, has anyone been using NIH Image or Imagej to do time-lapse } in color? } } I am new to this listserv, so my apologies if I'm raking over old } territory. I did do a search of the listserv archives and didn't find any } postings in this subject. Many thanks, Mary } =============================================== } Mary_Tyler-at-umit.maine.edu or mtyler-at-maine.edu } } Mary S. Tyler } Professor of Zoology } Dept. of Biological Sciences } University of Maine } Orono, ME 04469-5751 } FAX: 207-581-2537
Hello, I am working with apoptoic cells in plant tissues. I am using Evan's Blue as a test of viability and of apoptosis. Can anyone tell me what Evan's blue (EB) stains in the cell? I know that it can not enter a cell with an intact membrane but when it enters a damaged or dead cell what does the EB actually bind to? Ann
They are pretty good in locating info and fixing all sorts of items. They do mech, ion and diffusion pump rebuilds. Very good service and high quality.
gary g.
At 12:41 PM 7/30/2001, you wrote:
} We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I } have called Hitachi about information on the pumps, but they are having some } difficulty in locating any specs on them. Searching the Internet did not } give any information either. } } The pumps plug into what appears to be a standard 3-prong socket and the } pumps have information on the housing for 110 V, so I'm thinking that the } power requirements are fairly standard. What I don't know is what flow } rate, et cetera, to get for the replacement pumps. } } Does anyone know where I could locate the pump specs or could you recommend } some pumps that would work? And, in case you haven't figured it out } already, I'm not an expert on vacuum systems, so I may be going about this } the wrong way by looking at specs that really don't matter. If someone } could let me know what parameters are the important things to look for, I'd } be grateful. } } Thanks, } } -Norman Kay } AME Dept. } The Univ. of Arizona
Once again the MSA Education Committee is organizing Exhibitor Demonstrations on Tuesday, August 7 from 5:00 to 6:00pm in the Exhibit Hall. These mini-seminars or tutorial demonstrations are held in the booths of the participating companies after the Hall is closed to non-participants at 5:00 pm.
Signup sheets with titles and descriptions are at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA Education table soon, since the demonstrations get filled up quickly!
Here's a list of participating Exhibitors and titles:
Asylum Research Force Measurements 101 - Featuring the New, Low Noise BioLever
Carl Zeiss, Inc., Microscopy & Imaging Systems Zeiss LSM 5 PASCAL Personal Confocal Microscope Features and Capabilities.
Digital Instruments/Veeco Metrology Group Recent Advances in Throughput, Resolution, Sensitivity, and Functionality with Atomic Force/Scanning Probe Microscopy
Evex Analytical Video SEM
FEI Company High Resolution STEM Imaging on a TEM
Gatan, Inc. Fast Mapping with Hi-Speed EELS Spectrum Imaging
Hamamatsu Photonic Systems Hamamatsu Introduces IEEE1394
Hitachi Scientific Instruments Hitachi's NEW WxB Filter System on the S-4700 FESEM
IATIA Group Optical Phase Microscopy with QPm: Quantitative Imaging and Conventional Visualization Analogs
IXRF Systems, Inc. SEM-EDS Integration: The Future of Microanalysis!
Micro Photonics, Inc. Non-Destructive 3D Microscopy Using X-Ray Microtomography
Molecular Imaging PicoSPM and In Situ Imaging Under Controlled Environments
Olympus America, Inc. Motorized Microscopy - Making Your Life easier
Polaroid Corp. Polaroid's Digital Microscope Camera and Simple PCIp Software
Princeton Gamma Tech, Inc. "What is That PTS I Hear So Much About?"
Q3DM, Inc. Automating High Resolution Microscopy: Addressing the Challenges of High Throughput Fluorescence Microscopy for Cell-Based Assays
Quesant Instruments Corp. Quesant Instruments New Metrology System
Soft Imaging System Corp. Digital Imaging on Your TEM: Everything You Wanted to Know But Were Afraid to Ask.
South Bay Technology, Inc. #1-Sneak Preview of New Techniques and Tool Sets for the D500i DIMPLER #2-Ion Beam Sputter Deposition & Etching (IBS/e) #3-Dedicated Low Energy Ion Milling
Ted Pella, Inc. New Methods of Microwave-Assisted Processing for Electron Microscopy
TSL (EDAX booth) Phase Identification and TEM Tool
See you in Long Beach!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We are hoping someone out there, preferably within Australia, may have an ultramicrotome within their facility that has become redundant or obsolete (preferably an Ultracut E, Ultracut s or a Reichert OMU 3). If so, please reply ASAP.
We plan to buy CCD cameras for our two TEMs, JEM-2000 EXII and JEM2010F. The models on our final (almost) list are Gatan 692 (1K by 1K) for 2000Ex and 795.201.2 (2K by 2K) for 2010F
We use 2000 EX for routine IC failure analysis and 2010F for HR and EDS. Both CCDs are quite expensive. We will appreciate any user experience on both modles or any suggestions on lower cost ones.
You will be responsible for operating a transmission electron microscope (TEM), scanning electron microscope (SEM), taking and processing images. Responsibilities include collecting, processing, embedding, and sectioning tissues in support of electron microscopy services. You will keep records in compliance with SOP's and GLP's and maintain laboratory instruments in accordance with SOP's, troubleshooting equipment problems as needed. You will process samples in a timely manner according to SOP's. You will maintain and keep the laboratory clean and organized and ensure that instrumentation is cleaned, calibrated, and properly maintained according to facility SOP's. You will ensure that laboratory computer equipment operates smoothly and work with computer support to trouble-shoot network and software problems. You may also act as Pathology monitor for assigned Drug Safety studies. You will tabulate and report Imaging data to the laboratory supervisor and work with the laboratory supervisor to ensure the accuracy and completeness of all Pathology data presented in the final report to the FDA and sent to archives. You will assist in writing SOP's and appropriate forms to ensure compliance in all GLP studies and to maintain records for all studies that are non-GLP or in the spirit of GLP. You may also assist investigators with (SEM) and digital imaging needs. You will perform additional departmental and facility support functions such as (but not limited to) inventory, scheduling, or purchasing.
A Bachelor's degree is required in Biology, Biochemistry, or Chemistry or a related field with 7 years relevant experience or a Master's degree with 5 years relevant experience. 5-7 years experience working in transmission and scanning electron microscopy, in an industrial, hospital, or highly specialized academic setting is also required. Candidate should have strong communication and computer skills.
David W. Pereira Wyeth-Ayerst Laboratories 641 Ridge Road Chazy, NY 12921 Tel. (518) 846 6464 Fax. (518) 846 6345 email pereird-at-war.wyeth.com
Evans Blue will spontaneously react with albumenoids to form a covalent linkage. Under those circumstances, the dye becomes fluorescent (red).
Evans Blue has been used to tag albumen (1:1). Then it is dialyzed to remove unbound dye. Finally the tagged albumen is injected into an animal vascular system. It is used this way to test for sites of extravasation following injury or as an injection to test for blood volume. It is NOT known to be hazardous (I believe).
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Ann P. Ketter } Sent: Monday, July 30, 2001 8:17 PM } To: listserver microscopy } Subject: LM- what does evan's blue stain? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I am working with apoptoic cells in plant tissues. I am using Evan's } Blue as a test of viability and of apoptosis. Can anyone tell me what } Evan's blue (EB) stains in the cell? I know that it can not enter a } cell with an intact membrane but when it enters a damaged or dead cell } what does the EB actually bind to? } Ann } } }
Wyeth-Ayerst Research, the pharmaceutical division of American Home Products, is seeking an experienced and responsible microscopy technician at its Drug Safety facility in Chazy NY. This is a new, state of the art facility located just minutes from Lake Champlain and the Adirondack Mountains. Although located in rural setting it is only an hours drive to Montreal, Quebec and Burlington Vermont. You will be responsible for operating a transmission electron microscope (TEM), scanning electron microscope (SEM), taking and processing images. Responsibilities include collecting, processing, embedding, and sectioning tissues in support of electron microscopy services. You will keep records in compliance with SOP's and GLP's and maintain laboratory instruments in accordance with SOP's, troubleshooting equipment problems as needed. You will process samples in a timely manner according to SOP's. You will maintain and keep the laboratory clean and organized and ensure that instrumentation is cleaned, calibrated, and properly maintained according to facility SOP's. You will ensure that laboratory computer equipment operates smoothly and work with computer support to trouble-shoot network and software problems. You may also act as Pathology monitor for assigned Drug Safety studies. You will tabulate and report Imaging data to the laboratory supervisor and work with the laboratory supervisor to ensure the accuracy and completeness of all Pathology data presented in the final report to the FDA and sent to archives. You will assist in writing SOP's and appropriate forms to ensure compliance in all GLP studies and to maintain records for all studies that are non-GLP or in the spirit of GLP. You may also assist investigators with (SEM) and digital imaging needs. You will perform additional departmental and facility support functions such as (but not limited to) inventory, scheduling, or purchasing.
A Bachelor's degree is required in Biology, Biochemistry, or Chemistry or a related field with 7 years relevant experience or a Master's degree with 5 years relevant experience. 5-7 years experience working in transmission and scanning electron microscopy, in an industrial, hospital, or highly specialized academic setting is also required. Candidate should have strong communication and computer skills. Interested individuals should send their resumes, including a description of their expertise and references to:
David W. Pereira Wyeth-Ayerst Laboratories 641 Ridge Road Chazy, NY 12921 Tel. (518) 846 6464 Fax. (518) 846 6345 email pereird-at-war.wyeth.com
At 2:24 PM -0400 7/30/01, Don Gantz wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************** Hi Donald,
Much would depend on the formulation of araldite used. Like Epon, et al, it can be harder or softer if the ratios of components is varied. My experience is that, keeping relative hardness in mind, it behaves pretty much like epon. I'm sure others, with more experience working with it will have more info on this.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I expect that no two vacuum engineers would agree on the ideal pump size for a particular system: to wit the tiny pumps used by JEOL and the huge twin pumps used on most Hitachi scopes. Mechanical (also called rotary and fore pumps) are purchased largely on three figures: ultimate vacuum, pumping speed / volume (expressed as a volume over a time) and price. A small pump is cheaper to buy and uses less electricity, they tend to be quieter running (depending on make), larger pumps are the converse in properties but they pump down faster especially with new film in a TEM and when evacuating a larger space rather then a small airlock. In either case you must opt for a two-stage pump. Direct drive types are less trouble since belts break more frequently than couplings. Couplings become noisy and so warn the user that they need attention, but belts break with no warning. (It pays to inspect belts every three months or so, cracks on the inside of a belt indicated that a replacement is needed). Belt driven pumps run a little slower and live a little longer. Having said that, I would buy a direct drive pump one size larger than the reasonable minimum required. I think that 4cubic meters/ hour is a reasonable minimum for most EMs and so a 6 might be a good size. A 2 would work, but encourage an awful lot of long coffee breaks and a ten is a bit of overkill. I don't know how the Hitachi twin pumps share the work load, but it may not be easy to combine the pumping circuits, so you may need to retain two somewhat smaller pumps (2x 3 or 4 m3/h). Over the years I have used pumps of numerous makes and while they all work and most are quiet reliable, I would get a Leybold or an Edwards because they seemed to run quietest and achieved, for a given size pump the best ultimate vacuum. But these things may have changed in recent times. Perhaps other listers have an opinions on pumps performance. However, I should also declare my bias: ProSciTech is an agent (in Australasia) for Edwards pumps. Happily I have predilections for Edwards. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, July 31, 2001 5:42 AM, SEM Machine [SMTP:SEM-at-ACATC.AME.Arizona.edu] wrote: } } } We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I } have called Hitachi about information on the pumps, but they are having some } difficulty in locating any specs on them. Searching the Internet did not } give any information either. } } The pumps plug into what appears to be a standard 3-prong socket and the } pumps have information on the housing for 110 V, so I'm thinking that the } power requirements are fairly standard. What I don't know is what flow } rate, et cetera, to get for the replacement pumps. } } Does anyone know where I could locate the pump specs or could you recommend } some pumps that would work? And, in case you haven't figured it out } already, I'm not an expert on vacuum systems, so I may be going about this } the wrong way by looking at specs that really don't matter. If someone } could let me know what parameters are the important things to look for, I'd } be grateful. } } Thanks, } } -Norman Kay } AME Dept. } The Univ. of Arizona } } } }
There is no micro user onboard our research ship, and you are inundating our inmarsat connection with your email list. Please take care of this as soon as possible please,
Thank you,
Jeff Turmelle Sys Admin, R/V Maurice Ewing Lamon-Doherty Earth Observatory of Columbia University jefft-at-ldeo.columbia.edu
Question: I want to know the exact meaning of terms defocus,underfocus,focus,infocus and overfocus how they are classified for a particular object.
I would appreciate receiving reply in this regard
Dear Mullapudi Srinvas, For an ideal lens, all the electrons leaving a point on a specimen will be brought together in a point in a plane--the focal plane. When the lens currents are adjusted so that this plane is coincident with the detection system (film, CCD, etc.), the image will be in exact focus. (N.b., focus is a property of the image and is the same regardless of the particular object, although different points in thick objects can have different focal planes.) The distance between the focal plane and the detector plane is the amount of defocus--positive for overfocus (focal plane above the detector plane) and negative for underfocus. If you look at how information from the specimen is conveyed to the image as a function of spatial frequency--i.e., how the large and small features appear--you will find that there is a curve which defines the relationship between the amplitude of the wave of electrons leaving the specimen and that of the wave incident on the detector. This curve is the contrast transfer function (CTF), and it is of the same sign for frequencies lower than a certain value, then it changes sign and oscillates at higher frequencies. The amount of defocus which produces a CTF extending to the largest frequency before it changes sign gives the condition called Schertzer (sp?) focus. This condition results in images in which easily interpretable information extends to the highest resolution possible. Images are often taken somewhat underfocus, since that condition causes interference fringes to occur at edges due to the sign reversal of the CTF and enhances their visibility. This is often useful for images of biological specimens. I hope this answers your questions. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
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The Microscopy Lab at KU is getting a new SEM at the beginning of October. Consequently our current SEM, a Hitachi S-570 needs to be moved out before that time. If anyone needs an older SEM please contact me. Thank you Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id NAA27436 for dist-Microscopy; Tue, 31 Jul 2001 13:32:17 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id NAA27433 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 31 Jul 2001 13:31:47 -0500 (CDT) Received: from mail2.lsil.com (mail2.lsil.com [147.145.40.22]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id NAA27426 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 31 Jul 2001 13:31:34 -0500 (CDT) Received: from mhbs.lsil.com (mhbs [147.145.31.100]) by mail2.lsil.com (8.9.3+Sun/8.9.1) with ESMTP id LAA03065; Tue, 31 Jul 2001 11:28:44 -0700 (PDT) Received: from [172.21.35.125] by mhbs.lsil.com with ESMTP; Tue, 31 Jul 2001 11:28:02 -0700 Received: by exccolorado.co.lsil.com with Internet Mail Service (5.5.2650.21) id {P0GAJR2S} ; Tue, 31 Jul 2001 12:22:29 -0600 Message-Id: {BC3CE0375D29D51199A400508B66CA6C367E5C-at-exc-snake.co.lsil.com} Microscopy-at-sparc5.microscopy.com
Lee,
There is a substance called choline hydroxide (Sigma Aldrich) that has 10,000:1 selectivity (Si:SiO2). Dilute 25 ml. choline with 65 ml of DI water at 90 degrees C (must be maintained accurately). Depending on the thickness of your poly, it should take 1 - 5 minutes to remove the poly and keep the GOX intact. Hope this helps. Hey, if you know Yih-Shung Lin and he is still there at TSMC tell him that John Staman said "Hi".
Regards,
John Staman, LSI Logic
-----Original Message----- } From: "TCLEEG-at-tsmc.com.tw"-at-sparc5.microscopy.com [mailto:"TCLEEG-at-tsmc.com.tw"-at-sparc5.microscopy.com] Sent: Tuesday, July 31, 2001 6:35 AM To: Microscopy-at-sparc5.microscopy.com
I am in search of SEM services for small metallic medical products in the Minneapolis or St. Paul area in Minnesota. If there are any suggestions, I would appreciate them. Thank you. Dan May IntraTherapeutics dmay-at-iti-med.com {mailto:dmay-at-iti-med.com}
Dear Norman, The Cutevac 160VP is a 160 l/min direct-drive rotary pump. It plugs into any 110v socket. Any direct-drive single or double stage vacuum pump with the same capacity should work. BTW, what are you doing with the old pumps? I have seven of them running in my lab and the oldest have been running continuously for 19.5 years. I love them, fix them myself when they leak oil and would love to get some more. At 12:41 PM 7/30/01 -0700, you wrote: } } We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I } have called Hitachi about information on the pumps, but they are having some } difficulty in locating any specs on them. Searching the Internet did not } give any information either. } } The pumps plug into what appears to be a standard 3-prong socket and the } pumps have information on the housing for 110 V, so I'm thinking that the } power requirements are fairly standard. What I don't know is what flow } rate, et cetera, to get for the replacement pumps. } } Does anyone know where I could locate the pump specs or could you recommend } some pumps that would work? And, in case you haven't figured it out } already, I'm not an expert on vacuum systems, so I may be going about this } the wrong way by looking at specs that really don't matter. If someone } could let me know what parameters are the important things to look for, I'd } be grateful. } } Thanks, } } -Norman Kay } AME Dept. } The Univ. of Arizona } Regards adn good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi } } We plan to buy CCD cameras for our two TEMs, JEM-2000 EXII and } JEM2010F. The models on our final (almost) list are } Gatan 692 (1K by 1K) for 2000Ex and 795.201.2 (2K by 2K) for 2010F } } We use 2000 EX for routine IC failure analysis and 2010F for HR and } EDS. Both CCDs are quite expensive. We will appreciate any } user experience on both modles or any suggestions on lower cost ones. } } Regards, } Tan-Chen Lee
Dear Tan Chen Lee!
It would be useful to contact You directly to avoid direct advertisement on this mail list. Please send Your e-mail adress to the adress specified below.
My experience is that belt-drive pumps last a lot longer than direct, I have a couple of JEOL ones that have run more-or-less continuously for almost 30 years, with annual oil changes, frequent belt inspection and replacement as Jim says, and one replacement of the vane springs! Direct-drive ones don't seem to be able to do anything like that.
They are also quieter, particularly if the belts are sprayed occasionally with belt dressing.
Gimme belt-drives every time.
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I don't remember the procedure, but you may remove PERCOLL after purification by centrifugation at some particular acceleration (g). It's clearly stated in the PERCOLL Instruction Manual. If you need real data, I may check. I was using negatively stained mitochondria for TEM after PERCOLL purification and did not noticed any PERCOLL particles in the preparation, so it's not a problem if you remove PERCOLL properly after purification.
Sergey.
At 06:54 PM 7/26/01 -0600, you wrote: } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good } luck. Sergey } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Many thanks to everyone who replied. I now have some idea of where I need to go from here.
To answer a few of the questions that came up, the problem was that before the SEM was donated to us, I believe it was not serviced for a while nor was it handled very gently during transit. I just started working with the unit and recently found out that when it was delivered, it was left at the wrong building & sat on the loading dock for a few days - during monsoon season. So, needless to say, this unit has had a rough life. We've had it serviced & the Hitachi tech said that the pumps would most likely need to be replaced soon, but I should have questioned more into why she thought that. The pump failed a little over 3 months after the service, so I guess she was on to something. I'm also learning as I go, since there isn't anyone here to teach me, which I'm sure has contributed to the early demise of the pump.
What happened to the pump was that one morning when I came in, there was a puddle of oil around the pump, so something obviously gave out during the night (all those years of schooling have taught me a thing or two...). Of the two pumps, the one that died was the "stronger" one, since we already knew that the other pump was a bit weaker. I called Hitachi & they told me that they didn't have a rebuild kit, per se, but that they could send a list of parts - no guarantees on if they were still available, though. Based on my past experience with trying to find parts for our SEM, I'm pretty sure very few of the parts are still available. I did give the pump to our machine shop, which has done a bit of work on vacuum pumps, and they rebuilt it as best they could, but it wasn't enough, since I cannot get a high vacuum now.
I should mention, though, that a Hitachi tech did contact me after I submitted the initial post & was very helpful on getting me some information on pumps that could replace our unit. Based on what he said, and also from replies here, it appears that we need something that can do about 9 cfm. I also plan on contacting a few of the companies suggested here on costs to rebuild the pumps, since they may be salvageable.
I am trying to stain some formaldehyde preserved (no osmium) fungal hyphae so I can visualize them after embedding in LRWhite.
Any suggestions? Tryphan blue works until you transfer the hyphae to LRWhite, which extracts all color. I have oriented the hyphae in small blocks of agar, but the small size of the hyphae makes it difficult to see them if I stain the agar, say with Sudan black as was once suggested years ago.
We have purchased Gatan Model 600 Dual Ion Mill for use in the preparation of foils of geological material for TEM applications. Since this unit was purchased used from a government lab it was delivered without technical support and with incomplete documentation, leaving many unanswered questions. We are having problems maintaining high vacuum (the diffusion pump shuts off) and cannot find the source of the problem. Help from anyone familiar with this device would be greatly appreciated.
We are attempting to thin quartz arenites and have spent the majority of our time (when the unit is operational) in an effort to optimize the thinning process. Any input in that regard would also be welcome.
Thanks in advance for any information you can provide.
Kevin Macke Bowling Green State University Department of Geology Bowling Green, OH Email: macke-at-bgnet.bgsu.edu
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dlongerbeam-at-captusnetworks.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 31, 2001 at 19:24:09 ---------------------------------------------------------------------------
Email: dlongerbeam-at-captusnetworks.com Name: Donald Longerbeam
Organization: none
Education: Graduate College
Location: Roseville Ca.
Question: I was trained as a molecular biologist and did work with various microscopes over the years. However I have been out of the field for 25 years. I now have a son who I wish to introduce to the microscopic world. Who is making a good and reasonable microscope at this time. Olympus and Zeiss are certainly good - but they are too expensive. I am willing to spend up to $1000 apiece for a good binocular dissecting microscope and a good general higher power scope - . Thanks. Donald Longerbeam.
My usual golf partners won't be able to make it to M&M this year. Brian Cook and I are looking for two others to pair up with.
Thanks, Lucille Giannuzzi
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
We've been doing some hot stage environmental SEM (ESEM) work with aluminium-silicon alloys (up to 650°C), and we get some interesting contrast enhancement features as the temperature is increased. We are trying to discount various parameters and it seems that hot stage high vacuum (conventional) SEM would help determine if the effect has something to do with the gas in the chamber.
Will a conventional Everhart-Thornley (E-T) detector be able to magange this, or will contamination and out-gassing be a problem? Also I'm aware that unlike the gaseous secondary electron detector (GSED) in the ESEM, the E-T detector is sensitive to light, will this be a problem too? If it does not work, will it simply not give a good image, or is it likely to damage the dectector? Finally if anyone has tried this technique and it worked, any tips for good imaging?
Thank-you,
Simon Hogg
--
Dr. Simon Hogg Thixoforming Research Group University of Sheffield Department of Engineering Materials Sir Robert Hadfield Building Mappin Street Sheffield S1 3JD Tel. 0114 222 5934 Fax. 0114 222 5943
Where's the connectioln between this list and golf partner problems. Hard to see for me in Sweden. There should be other channels for personal stuff like this.
Yours sincerely
Per Hörstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Umeå S-90187 Umeå Sweden
Hi Dan, The Imaging Center in the College of Biological Sciences houses a new Hitachi S3500N SEM. The instrument is a variable pressure system equipped as follows:
*** Hitachi S-3500N, variable pressure SEM ***
- Standard stage with motor driven X & Y axes.
- Robinson Backscattered Electron Detector (BSE).
- Hitachi Hi-Mouse software.
- Infrared ChamberScope.
- Photo CRT Unit, recording camera.
- Ethernet interface.
*** EMITECH K-1150 CRYOGENIC SYSTEM *** - http://www.emitech.demon.co.uk/
*** EDAX Phoenix X-Ray Microanalysis System ***
- Windows NT Operating System
- Super Ultra-Thin Window 10mm LN-cooled Detector
- EDX Fast Mapping, EDX Quant Mapping, EDX Line Scan, Spectral Mapping/Data Recovery, SEM Quant Package.
We also house a large assortment of TEM and LM instrumentation and preparation equipment. We can provide service or teach you how to use the instrument. Feel free to check out our web site and contact either Mark Sanders (msanders-at-biosci.umn.edu) or Gib Ahlstrand (giba-at-puccini.crl.umn.edu) with any questions or to arrange a demonstration. Cheers, Mark **************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus Imaging Center St. Paul, MN 55108 23-35 Snyder Hall ph: 612-624-3454 1475 Gortner Ave. fax: 612-624-1799 http://biosci.umn.edu/imagingcenter
} From: "May, Dan " {DMay-at-iti-med.com} } Date: Tue, 31 Jul 2001 14:04:54 -0500 } To: "'Microscopy-at-MSA.Microscopy.com'" {Microscopy-at-sparc5.microscopy.com} } Subject: SEM Services in Minneapolis/St.Paul Minnesota Area } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am in search of SEM services for small metallic medical products in the } Minneapolis or St. Paul area in Minnesota. If there are any suggestions, I } would appreciate them. } Thank you. } Dan May } IntraTherapeutics } dmay-at-iti-med.com {mailto:dmay-at-iti-med.com} } } }
I am looking for a suitable plastic embedding material which should have the following characteristics:
- applicable for light microscopy - not be autofluorescent - used for immunocytochemistry, i.e. tissue should retain immunogenicity - soft enough to cut 20 micron or thicker sections easily - ability to embed large pieces e.g. entire rat lung - opacity to allow identification of parts of embedded tissue (this excludes paraffin)
I hope I am being realistic and there is a product out there to meet these criteria. This group has never let me down.
Thank you in advance. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
Relax, There has been a precedent in the past for using this forum for arranging social events at the M&M meeting. There are no other forums for reaching the large number of participants that go to the meeting. It has never gotten out of hand before and someone shouldn't be criticized for doing it. It is at social functions that sometimes the most valuable scientific work at a conference gets done.
If you do attend one of the M&M meetings, look me up; I'll buy you a beer.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Per Hörstedt [mailto:per.horstedt-at-pathol.umu.se] Sent: Wednesday, August 01, 2001 7:05 AM To: microscopy-at-sparc5.microscopy.com
Where's the connectioln between this list and golf partner problems. Hard to see for me in Sweden. There should be other channels for personal stuff like this.
Yours sincerely
Per Hörstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Umeå S-90187 Umeå Sweden
A quick note about replacing rotary pumps on Hitachi Microscopes. The microscope provides 100 VAC (Japan Voltage) for operation. Hitachi Instruments service uses Edwards pumps as replacements in the USA, and they seem to be OK at this low voltage. Other brands must be checked.
Dunniway Stockroom doesn't like to try to rebuild the Hitachi brand pumps because they find it hard to get parts. So they don't deal with them. Mary must know a secret about parts for rebuilding them.
Direct drive pumps operate at higher RPM and wear out sooner than belt drive pumps.
Ronald Vane XEI Scientific Redwood City, CA
-----Original Message----- } From: Mary Mager {mager-at-interchange.ubc.ca} To: SEM Machine {SEM-at-ACATC.AME.Arizona.edu} Cc: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
POSTDOCTORAL POSITION in Quantitative Electron Microscopy
A postdoctoral position is available in the area of quantitative TEM/TED of materials starting September 1, 2001. The research project focuses on developing quantitative methods of solving structures from diffraction patterns and HREM images. A good background in computer programming in fortran or C is required, as well as an understanding of dynamical diffraction theory. Prior experience in HREM or Direct Methods would be an advantage.
For further information or applications email Professor L. D. Marks at ldm3-at-risc4.numis.nwu.edu
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Montana Tech's Dept of Metallurgical and Materials Engineering is in need of replacing its SEM/EDX system and is looking for an economical used system. An SEM w/o EDX can be considered since the EDX on our current system can likely be attached. Parties interested in divesting their SEM/EDX system should contact Dr. Courtney Young by e-mail as soon as possible at cyoung-at-mtech.edu or by phone at 406-496-4158 to let him know what the system is, what its condition is, its age, if there is an EDX attached, and what the cost will be to get it delivered to Montana Tech in Butte MT.
We are long time users of the Metamorph image analysis software package from Universal imaging. Our sales rep had told us they were coming up with a networkable edition so that we could buy a licence for a number of copies that would allow users on our campus to install the software on their own lab computers and it would run as long as the number of copies running at any moment was not more than the number of licences we had. It would check this over the internet backbone. We are now told that they have either abandoned this idea or never considered it depending on who and when we talk to them. It is my understanding that ImagePro does have such a capability and we are considering switching over to it. Is there any one out there with experience with this set up? Does it work? I am particularly interested in a solution in which the software runs on the local computer and not on a server so that we would not have to waste time transferring files, etc. over the net. I assume it would be a lot faster to have the software simply look for the licence over the net. If i am wrong about this, I would be happy to be educated on this matter also.
I would also welcome comments comparing the ease and versatility of ImagePro vs. Metamorph.
Thanks, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Fall 2001 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section E2)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A fourteen week, Fall 2001 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered one evening per week (Thursdays) starting at 5:30 PM. Classes will begin on September 6 and end on December 20, 2001.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $92 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Have you looked at the MSA Surplus Equipment pages
http://www.msa.microscopy.com/SurplusEquipment/
Nestor Your Friendly Neighborhood SysOp
} } } Montana Tech's Dept of Metallurgical and Materials Engineering is in need of } replacing its SEM/EDX system and is looking for an economical used system. } An SEM w/o EDX can be considered since the EDX on our current system can } likely be attached. Parties interested in divesting their SEM/EDX system } should contact Dr. Courtney Young by e-mail as soon as possible at } cyoung-at-mtech.edu or by phone at 406-496-4158 to let him know what the system } is, what its condition is, its age, if there is an EDX attached, and what } the cost will be to get it delivered to Montana Tech in Butte MT.
-- =========================================== Dr. Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4289 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
Recently, we started experiencing a problem when using PC disks (1.8MB and Zips) in Macs. We are able to access and write to the PC disks but can not eject the disks without rebooting the Mac. We get a message indicating that the file is in use (even though we have quit the program) and a second message indicating that file sharing could not be enabled. I suspect that this may be due to the disks being formatted on the latest PC operating system. Does anyone know of a fix? Thanks.
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
John, Use Mac formatted disks on the PC and use Conversions Plus from DataViz. I have never had a problem doing anything this way. What works best is to use PC Formatted Zips and then quick format them in the PC to the Mac Format. I have been doing this for about 4 years and it works great. I never lose the long formatted names going to either platform.
The one thing that you do have to take care of is to make sure that you close the directory structure on the Mac before you eject it and carry it to the PC. If you modify it on the PC after you ejected it on the Mac with the directory structure open and then go back to the Mac, your information can get lost. (I think that I got that sequence right --I may have gotten it backwards.) Whatever you do, close the file structure before ejecting the disk from the Mac.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Wednesday, August 01, 2001 2:29 PM To: Microscopy-at-sparc5.microscopy.com
Recently, we started experiencing a problem when using PC disks (1.8MB and Zips) in Macs. We are able to access and write to the PC disks but can not eject the disks without rebooting the Mac. We get a message indicating that the file is in use (even though we have quit the program) and a second message indicating that file sharing could not be enabled. I suspect that this may be due to the disks being formatted on the latest PC operating system. Does anyone know of a fix? Thanks.
John B.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
I am looking for a suitable mounting material to mount small fossil bone samples. The sample extraction method requires the material to be optically clear and will allow some of the mounting to be mixed with the sample. So it would be desirable for the mounting material not to interfere with the elemental isotopic ratio that exists in the sample. Phosphorus compounds are of most concern. Any suggestions would be appreciated.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
I am considering the purchase of a used Zeiss Standard 25 over the web. I will not have the opportunity to inspect the scope and can judge it from photographs only. It has 5 plan acro objectives (2.5, 10, 25, 40, and 100 (oil)), but otherwise seems to be the "base" configuration with no optional components.
I have searched the web for information on this scope and have come up empty-handed (empty-screened?) I would like to know when they were produced, where they fell in the Zeiss product line, what the "successor" to the Standard 25 was or is, where I might find an instruction manual or old product catalog that describes optional configurations, and any other information about the scope.
Would anyone venture an opinion on the quality of this scope or its value? The asking price is $1500. It is said to be in good condition.
Thanks very much, Ed Bachmann
Ed Bachmann Odum Institute for Research in Social Science Manning Hall CB 3355 University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3355 (919) 962-0512
We have had an identical problem with PC formatted Jaz and Zip disks. It appears, in our case, there is a conflict with Norton Anti-virus, Apple's PC Exchange, and PC-formatted Iomega media. If we disable Norton Anti-virus Auto Protect before inserting the Zip or Jaz disk, the problem goes away. This is obviously not a permanent solution. Let me know if you come up with a better solution.
Rod
At 01:28 PM 8/1/01 -0500, John J. Bozzola wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
----------------------------------------------------------------------------------------- Rodney McCabe MST-8 : Structure Property Relations MS-G755 Los Alamos National Laboratory Los Alamos, NM 87545 Phone: (505) 665-3289 -----------------------------------------------------------------------------------------
I have read with fascination Debbie¹s description of the MSA Facility Workshop, and it sounds great. However, for those of you involved in disease and pathology microscopy, the session on Emerging Diseases (unfortunately on the same day) will also pique your interest. We have internationally recognized speakers, including one from Australia and two from the Centers for Disease Control and Prevention. Also there are an AIDS and opportunistic pathogen expert from Geo Wash U and an immunopathologist and transplantation expert from Duke. There are talks on Ebola virus, use of EM to detect new viruses, prion diseases, agents of biowarfare, and others. I hope many of you can join us.
Sara
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I need to order a compound microscope to look at thick sections from plastic embedded plant material before thin sectioning. Could someone suggest one that cost less than $2,500.00?
It seems that you already know the answer: If there is any light emitted, and at 650 C most materials would, then the photomultiplier tube, which is integral to the E-T detector would be overloaded. You can check that by increasing the multiplier tube amplifier from zero up, until it overloads or you reach your usual mid-range setting. Similarly, outgasing at high temperatures is a problem and I would be surprized if you could attain a vacuum sufficient for the ET detector - BS detectors are rather less sensitive to poor vacuum. One obvious way to minimize outgasing would be to have a very small area heated. My question is: How do you avoid heat damage within the chamber, including to the plastic (could be quartz) scintillator and light pipe? Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, August 01, 2001 6:43 PM, S.C.Hogg [SMTP:S.C.Hogg-at-sheffield.ac.uk] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } We've been doing some hot stage environmental SEM (ESEM) } work with aluminium-silicon alloys (up to 650?C), and we get some } interesting contrast enhancement features as the temperature is } increased. We are trying to discount various parameters and it } seems that hot stage high vacuum (conventional) SEM would help } determine if the effect has something to do with the gas in the } chamber. } } Will a conventional Everhart-Thornley (E-T) detector be able to } magange this, or will contamination and out-gassing be a problem? } Also I'm aware that unlike the gaseous secondary electron detector } (GSED) in the ESEM, the E-T detector is sensitive to light, will this } be a problem too? If it does not work, will it simply not give a good } image, or is it likely to damage the dectector? } Finally if anyone has tried this technique and it worked, any tips for } good imaging? } } Thank-you, } } Simon Hogg } } } } -- } } Dr. Simon Hogg } Thixoforming Research Group } University of Sheffield } Department of Engineering Materials } Sir Robert Hadfield Building } Mappin Street } Sheffield } S1 3JD } Tel. 0114 222 5934 } Fax. 0114 222 5943
I acquired some EELS spectrum of diamond-like BCN phase and would like to analyze them. Does anyboday know if a standard spectrum of cubic BCN (theoretical or experimental) can be found somewhere?
Thanks in advance for your help!
James
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
Thank you to all who responded to my previous posting. Like most of the respondents, we thought the most likely problems were a faulty thermal switch or a lack of cooling water, so we checked those first (I forgot to include that detail. sorry). We managed to trace the problem down to the thermocouple gauge today. Apparently the optical switch that triggers the 20-amp relay after the preset vacuum is reached is malfunctioning. After discussing the problem with the people at Gatan and discovering the cost of a new gauge (approximately $700-1500, depending on the source), we went the low buck route and bypassed the TC gauge by running a jumper from the green wire on the "B" switch to the red wire on the relay. Now the "B" switch controls the diffusion pump heating element directly and things are working fine. Hopefully this will save some time and frustration for anyone out there who ends up with a similar problem.
Our last problem is finding the best way to thin the quartz samples. The procedure I have followed in the preparation of the grids prior to ion milling appears at the end of this posting.
My question is this: has anyone found a particularly advantageous setup for the Gatan in milling a material such as a quartz arenite? Specifically, what combinations of gun angles and voltages work best?
(For those not familiar with the Gatan 600, there are two guns firing towards the top of the sample. The guns are linked and can be set so both are at 20 degrees, or some combination such that gun A is at 20 degrees - x degrees and gun B is at 20 degrees + x degrees (e.g. 25 & 15 degrees), to a maximum of 40 degrees and a minimum of 0 degrees. The current and amperage on both guns can be independently varied from 0-4 kV (the manual recommends 0.5 mA for most materials).)
Preparation of grids:
- prepared a standard (30 um) thin section using Crystal Bond and a Logitech thinner
- thinned by hand using diamond/resin polishing disks to approximately 20 um
- applied grids to section with epoxy, applied brass support rings to grid with epoxy. It looks like this:
I have a research group interested in looking at cross sections of coal char particles. The particles are quite porous and have been embedded in epoxy and cross sectioned. There is very little contrast in the SEM images as we are looking at carbon in carbon! In order to get a reasonable image I need a fairly large spot size (3nA, 15kV) which damages the resin which can be a large part of the field of view. I've tried C and Au coatings, the Au reduces the damage slightly but it is still very obvious, especially after a slow scan. I'm thinking a better conducting resin might help, or a more beam resistant resin or one that gives a high contrast compared to the char. Do such things exist? The resin cannot have particles in it (Ag, Cu etc) as a smooth background is required for image analysis. Any other suggestions on imaging such samples are welcome!
Thanks
Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
I am Marilena Re of ENEA Brindisi (Italy). I have spoken of this problem with my technicians, because we have two 600ion mill. The cause of the shut off of the diffusion pump should be due to the existence of electronic valve before, which shuts off the diffusion pump for protection when the flow of the water is not enough to cool it. So you should check it and control, for example, if it is dirty or if the flow of the water is enough. For your interest, I believe that you should ask to Gatan in order to have the manual, because it is very useful to solve many problems. Marilena
I am Marilena Re of ENEA Brindisi (Italy). I have spoken of this problem with my technicians, because we have two 600ion mill. The cause of the shut off of the diffusion pump should be due to the existence of electronic valve before, which shuts off the diffusion pump for protection when the flow of the water is not enough to cool it. So you should check it and control, for example, if it is dirty or if the flow of the water is enough. For your interest, I believe that you should ask to Gatan in order to have the manual, because it is very useful to solve many problems. Marilena
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We have purchased Gatan Model 600 Dual Ion Mill for use in the } preparation of foils of geological material for TEM applications. } Since this unit was purchased used from a government lab it was } delivered without technical support and with incomplete } documentation, leaving many unanswered questions. We are having } problems maintaining high vacuum (the diffusion pump shuts off) and } cannot find the source of the problem. Help from anyone familiar } with this device would be greatly appreciated. } } We are attempting to thin quartz arenites and have spent the majority } of our time (when the unit is operational) in an effort to optimize } the thinning process. Any input in that regard would also be welcome. } } Thanks in advance for any information you can provide. } } Kevin Macke } Bowling Green State University } Department of Geology } Bowling Green, OH } Email: macke-at-bgnet.bgsu.edu } } _________________________________________________________________ } Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
I am posting an advert for a vacancy we have for a food microscopist. Please forward it to anyone you know might be interested but does not subscribe this list.
Food Microscopist, Reading, UK
We at RSSL are looking for a Food Microscopist to join and add to our vibrant and hard working Microscopy team. Much of the work that fully occupies the current staff of 8½ is accounted for by our successful foreign body analysis service, where we identify foreign materials and particulate contamination, largely for the food and pharmaceutical industries. In addition, we have an established special microscopy business, a large part of which concerns food microstructure and we are looking to develop this business and expand it to move into different food systems.
If you are a self-starter, innovative, looking for an interesting and challenging role and believe you can make a positive contribution to our already successful team, we would very much like to hear from you. Ideally, you will have had a number of years practical experience of the application of various modes of microscopy to the investigation of different food systems and be educated to at least degree level. The Company is situated on the University of Reading campus within pleasant surroundings and can offer a competitive salary and benefits package, including contributory pension, private medical cover and a Company share-save scheme.
Please send your CV with a covering letter including your salary expectations to Jane Cook, Human Resources Officer, Reading Scientific Services Ltd, Lord Zuckerman Research Centre, PO Box 234, Whiteknights, Reading, RG6 6LA, UK or e-mail Jane.cook-at-rssl.com . RSSL is committed to providing equal employment opportunities.
Reading Scientific Services Limited is a leading provider of scientific research, analysis and consultancy to a wide range of clients from within the food, consumer goods, healthcare and chemical industries. As a Company, we are committed to scientific excellence, the provision of first class customer service and working to quality standards
******************************************************************** This e-mail is confidential and may contain privileged information. If you are not the addressee it may be unlawful for you to read, copy, distribute, disclose or otherwise use the information in this e-mail. If you are not the intended recipient please notify us immediately.
Reading Scientific Services Ltd, The Lord Zuckerman Research Centre, Whiteknights, PO Box 234, Reading. RG6 6LA.
i will be starting a new core lab. we will be buying a new microscope. i need input for a high resolution TEM with a film quality digatal camera system. anyone have any input? was thinking either FEI or JEOL. john hoffpauir
We have had a 250 Mark 3 since it was new in 1985.
It has been a rugged old SEM.
In the past few years parts are becoming increasingly difficult for even our LEO service engineers to obtain (thank goodness it's still on service contract).
It is a great SEM if you have large samples, need a lot of beam current and don't do a lot of high mag work.
The 4 quadrant BSE detector is a real gem.
We use ours as a back up to our JEOL 5800.
Sometimes I love to use it, so I can twist all those knobs.!
If you need any assistance that a well used user can supply, give me a call.
William T. Giles Sr. Electron Microscopist Met. Lab. Coordinator Henderson Technical Laboratory TIMET PO Box 2128 Henderson NV 89009 Ph: (702)566-4436 Fax: (702)564-9038 E-mail: Bill.Giles-at-timet.com } -----Original Message----- } From: Bart Cannon [SMTP:cannonmp-at-accessone.com] } Sent: Wednesday, August 01, 2001 3:22 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Cambridge S-250 SEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Everyone, } } A nicely mothballed Cambridge S-250 with many lovely accessories has } fallen into my lap. I am contemplating bringing it back from the dead. } } Is there anyone among you who is still delighted their S-250 who would } encourage me to proceed? } } Bart Cannon } Cannon Microprobe } Seattle }
1) I just talked to a gentleman at Fullam who told me he can longer obtain the Linde type 4A molecular sieves I've always used to keep my 100% ethanol dry. Does anyone out there have a source for these or, perhaps more likely, is there some great substitute for them that I'm not aware of?
2) Another quick query: can someone suggest a source for phosphorus pentoxide in which it arrives as larger crystals (as it seems it used to) instead of powder?
I appreciate the help I've had in the past from the listserver.
Thank you
Scott Robinson
Microscopy Suite, room B650J Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana IL 61801 217 265-5071 (office); 217 244-6219 (fax) sjrobin-at-uiuc.edu
Dave, you are using large spot size, so, I believe, you do not need high magnifications. I do not know what type of SEM you have, but I would suggest to try different tilt angles (it can increase contrast, so you can use lower current). Then you can try to use as low voltage as possible with your scope. And if your SEM has a possibility of frame averaging/integration, use this feature instead of slow scan.
If nothing works, try polishing your sections with soft clothes, it will create relief (but some distortion will occur, and image analysis could be less precise).
Vladimir
} G'day } } I have a research group interested in looking at cross sections of } coal char particles. The particles are quite porous and have been } embedded in epoxy and cross sectioned. There is very little } contrast in the SEM images as we are looking at carbon in carbon! } In order to get a reasonable image I need a fairly large spot size } (3nA, 15kV) which damages the resin which can be a large part of } the field of view. I've tried C and Au coatings, the Au reduces the } damage slightly but it is still very obvious, especially after a slow } scan. } I'm thinking a better conducting resin might help, or a more beam } resistant resin or one that gives a high contrast compared to the } char. Do such things exist? The resin cannot have particles in it } (Ag, Cu etc) as a smooth background is required for image } analysis. Any other suggestions on imaging such samples are } welcome! } } Thanks } } Dave } } } } } Dave Phelan } EM/X-Ray Unit } University of Newcastle } NSW 2308 } AUSTRALIA } Ph 02 4921 5667 } Fax 02 4921 7019 } emudp-at-mail.newcastle.edu.au } } }
I would like to know more about methods to adhere cell culture cells to both glass and plastic cover slips. Are there any alternatives to polylysine and does it work with plastic coverslips? I currently use a .001% aqueous solution of polylysine, but by the time I take the cells through all of the dehydrations, there are not many cells left. I probably need both more cells and I thought, better adherence thru a "stickier" cover slip. I am using various cells including Hela and several oral cancer cell lines. Any other methods? Your advice is appreciated.
Barbara Plowman Univ. of the Pacific School of Dentistry 2155 Webster St. San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu
We have several options available at Energy Beam Sciences that you may want to consider.
The first is the Technovit 9100, Methyl Methacrylate Kit. This kit is typically used to embed undecalcified bone. However, their is a monomer provided with the kit, which can be mixed with the plasticiser to vary hardness when embedding softer tissue.
Another option is the Technovit 7100, Glycol Methacrylate kit. Samples from lung to undecalcified bone are routinely processed with Glycol Methacrylate, therefore, hardness of the compund does not usually present an issue.
You can find these products on our web site at www.ebsciences.com. From the home page click on General Supplies. Under the heading of Product Information click on Light Microscopy Supplies. This will bring you to the page with the embedding kits listed.
I hope this helps.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: Judy Trogadis [mailto:TrogadisJ-at-smh.toronto.on.ca] Sent: Wednesday, August 01, 2001 9:11 AM To: Microscopy-at-sparc5.microscopy.com Cc: Judy Trogadis
Fellow Microscopists:
I am looking for a suitable plastic embedding material which should have the following characteristics:
- applicable for light microscopy - not be autofluorescent - used for immunocytochemistry, i.e. tissue should retain immunogenicity - soft enough to cut 20 micron or thicker sections easily - ability to embed large pieces e.g. entire rat lung - opacity to allow identification of parts of embedded tissue (this excludes paraffin)
I hope I am being realistic and there is a product out there to meet these criteria. This group has never let me down.
Thank you in advance. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
} I need to order a compound microscope to look at thick sections from } plastic embedded plant material before thin sectioning. Could someone } suggest one that cost less than $2,500.00? } } Thanks, Nancy
You can cut a zero off that price by shopping the Chinese imports intended primarily for the educational market. The quality is remarkably good, considering the prices; quite good enough for your purpose. You'll find a dealer list on the MICRO website (URL below).
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Barbara We use polylysine at 0.01% not 0.001%. See if this helps.
-----Original Message----- } From: Barbara Plowman [mailto:Bplowman-at-sfmail.dental.uop.edu] Sent: Friday, 3 August 2001 5:04 AM To: Microscopy-at-sparc5.microscopy.com
I would like to know more about methods to adhere cell culture cells to both glass and plastic cover slips. Are there any alternatives to polylysine and does it work with plastic coverslips? I currently use a .001% aqueous solution of polylysine, but by the time I take the cells through all of the dehydrations, there are not many cells left. I probably need both more cells and I thought, better adherence thru a "stickier" cover slip. I am using various cells including Hela and several oral cancer cell lines. Any other methods? Your advice is appreciated.
Barbara Plowman Univ. of the Pacific School of Dentistry 2155 Webster St. San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu
At 10:11 2/08/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Use dehydrated copper sulphate. Bake crystalline copper sulphate over a gas flame until it is a white powder. What we then do is to use dialysis tubing as a container so we have a "sausage" of anhydrous CuSO4 which we put in the 100% alcohol or acetaone bottle. The CuSO4 is insoluble in alcohol and it turns blue when rehydrated - a useful indicator of the presence of water............
} } Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
Myself and a few other folks are looking for a firm to service PGT systems in the central Massachusetts area, other than PGT. Any suggestions? Thanks in advance
Joseph M. Oparowski Materials Science Research - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have used anhydrous sodium sulfate for years to keep ETOH and acetone dry. It works fine. Just add about an inch to the bottom of a pint bottle of ETOH. Discard when you use all of the ETOH.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
Hi You All--
1) I just talked to a gentleman at Fullam who told me he can longer obtain the Linde type 4A molecular sieves I've always used to keep my 100% ethanol dry. Does anyone out there have a source for these or, perhaps more likely, is there some great substitute for them that I'm not aware of?
2) Another quick query: can someone suggest a source for phosphorus pentoxide in which it arrives as larger crystals (as it seems it used to) instead of powder?
I appreciate the help I've had in the past from the listserver.
Thank you
Scott Robinson
Microscopy Suite, room B650J Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana IL 61801 217 265-5071 (office); 217 244-6219 (fax) sjrobin-at-uiuc.edu
I have mouse ears that were glutaraldhyde fixed, post-osmicated, ethanol dehydrated, and embedded in Spurr. They were embedded so as to give a cross section through the modiolus. I am trying to get serial sections that are 2 microns thick and collected every 10 microns. The block face is approximately 7x4 mm. There is a lot of empty plastic within the sections that causes bubbles and folds. I have tried flattening the sections by using 0.5% to 1% benzoyl alcohol in water in the boat, or waving a stick dipped in acetone over the sections. I have tried FisherBrand SuperFrost slides and FisherBrand Premium microscope slides. I have very few sections out of hundreds that are relatively flat. I float them on a drop of water or in a puddle of water on the slide. I heat them on a hot plate at about 80-90° C, then transfer them to a slide warmer at 70° C. Any suggestions to improve flattening are desperately needed and all will be tried! Thanks.
Use dehydrated copper sulphate. Bake crystalline copper sulphate over a gas flame until it is a white powder. What we then do is to use dialysis tubing as a container so we have a "sausage" of anhydrous CuSO4 which we put in the 100% alcohol or acetaone bottle. The CuSO4 is insoluble in alcohol and it turns blue when rehydrated - a useful indicator of the presence of water............ Dear List, Does anyone know the free energies and/or partition coefficients for water in EtOH and CuSO4? I.e., if one puts 1 g of CuSO4 in 1 l EtOH what fraction of the residual H2O is extracted from the EtOH? TIA. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I would like some advice. I am doing FISH (Fluorescent In-Situ Hybridization)and have been unsuccessful in getting any staining. I am using DAPI counterstain, but have no protocol indicating how long to stain. The probe I am using is a dual probe (from Vysis).
If anyone is familiar with doing FISH (and successful!) and could get in touch with me with protocols they have used,including any problem areas, I would appreciate it!
You can contact me off the list server at sherwood-at-helix.mgh.harvard.edu.
Thanks!
Peggy Sherwood -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
----- Original Message ----- } From: "Marco Arienti" {marienti-at-tiscalinet.it} To: {"JHoffpa464-at-aol.com"-at-sparc5.microscopy.com} Sent: Friday, August 03, 2001 7:23 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hagin-at-iname.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, August 3, 2001 at 11:17:35 ---------------------------------------------------------------------------
Email: hagin-at-iname.com Name: hagin
Education: Graduate College
Location: City, State, Country
Question: Are there ROUTINE procedures to view dstrand or sstrand DNA in a resolution of 20 nm ? I have seen pictures of such macromolecules in the desired, and even better, resolution. My question intends to find out whether this pictures were obtained by a procedure requiring non-repeatable steps ? selection of one image out of hundreds ? trial and error ? and so on. Or, may be there are now procedures which can be easily taught, and which guarantee , given some miligrams of tissue, thousands of pictures of dna segments of hundreds of base-pairs long? If there are, could you please refer me to the literature which describes these procedures , or to the individuals who have the knowledge ? Sincerely Yona Hagin
the specimen on my desk is a polyurethane foam (about 100 pores/inch) covered with polypyrrole. I would like to get a ultramicrotome cut of it for TEM anlalysis. Can anybody suggest a preparation method? Which resin should I try for embedding and are there any other tips and tricks about this kind of material? I want to look at an unstained specimen and I would like to try a staining which will allow me to distinguish between the PU and Polypyrrol (and the resin, of course). Will RuO4 or OsO4 work with this combination?
Thanks for your input :)
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Laboratoire d'Analyse des Materiaux (LAM) Centre de Recherche Public - Gabriel Lippmann 162a, av. de la Faiencerie, L-1511 Luxembourg tel. +352-470261-503 fax +352-470261-549 e-mail: petra.wahlbring-at-crpgl.lu http://www.crpgl.lu/lam
Years ago I did some work on foams by TEM and OM. What you do depends on whether it is a closed cell, open cell foam, how rigid the foam is and the cell size. I had to work with fairly rigid closed cell PU foams. I used an Epon clone resin for embedding with no staining. Since only the broken cells at the edges would infiltrate, I chopped up the foam into small pieces so they could be wet with Epon. Try to concentrate the randomly oriented pieces in a small volume of Epon. In the TEM you have to search the thin sections for the structures you want to see. I was looking for particles in the cell walls and at struts.
It helps a bit to use Weck blades (Wecprep surgical blades) to make thin slices of the foam for a stereo microscopic view and a photograph of the structure you will see in the TEM. These blades are real real sharp razor blades (EM suppliers have them) and you can get some nice thin slices for doing OM using a vice and cutting the foam by hand. I had to do the cell size distributions of different PU foams using image analysis. Obviously the OM 'sections' had to be nearly as thin as the cell sizes.
In the TEM you can document the cell walls and struts if you chose your fields well. You can see the different types of struts junctions.
Hope this helps. My views are mine and not those of PPG Industries.
Paul Beauregard Sr. Research Associate PPG Industries Materials Analysis Lab Monroeville Technical Center Monroeville, PA 15146
-----Original Message----- } From: Petra Wahlbring [mailto:petra.wahlbring-at-crpgl.lu] Sent: Monday, August 06, 2001 7:58 AM To: microscopy-at-sparc5.microscopy.com
Dear listers,
the specimen on my desk is a polyurethane foam (about 100 pores/inch) covered with polypyrrole. I would like to get a ultramicrotome cut of it for TEM anlalysis. Can anybody suggest a preparation method? Which resin should I try for embedding and are there any other tips and tricks about this kind of material? I want to look at an unstained specimen and I would like to try a staining which will allow me to distinguish between the PU and Polypyrrol (and the resin, of course). Will RuO4 or OsO4 work with this combination?
Thanks for your input :)
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Laboratoire d'Analyse des Materiaux (LAM) Centre de Recherche Public - Gabriel Lippmann 162a, av. de la Faiencerie, L-1511 Luxembourg tel. +352-470261-503 fax +352-470261-549 e-mail: petra.wahlbring-at-crpgl.lu http://www.crpgl.lu/lam
We are currently in the process of decommissioning our VG HB501-ME0766 STEM. The instrument is in working order and is currently under service contract. Any interested parties should contact: Larry Murphy, Manager - Cabot Analytical Laboratories. Email: Lawrence_Murphy-at-Cabot-Corp.com. or phone (978) 670-6249.
I used to use Ted Pella's Medcast resin kit for my epoxy 'epon 812' clone material. I was forced to switch to Eponate 12.
Some background and for your information: Years ago, the problem with switching was that some suppliers said Epon 812 clone, identical to Epon 812 by NMR, identical by IR, etc.
A few years ago, I ran NMRs on Eponate 12, Epon 812 (original Shell material), and Medcast. They were identical by NMR. Now I know that NMR is not a quantitative technique for assay but it was a way to screen the different clones. I use Eponate 12 now. I have to use 70 degree for curing with Eponate 12 for some reason.
I ran one other supplier's material and it was totally different--totally different!!!
The original Shell(?) EPON 812 stuff I used as a check or reference material was and is much thicker than the material I buy and had bought recently.
Someday I will get and run other suppliers materials or kits. I may have to switch again! I suspect a lot of this material comes from the same chemical source.
Maybe someone out there has screened other material already and could send A SEPARATE EMAIL THREAD about their analysis of 'epon clones'?
Paul Beauregard Sr. Research Associate PPG Industries 440 College Park Drive Monroeville, PA 15146
-----Original Message----- } From: Petra Wahlbring [mailto:petra.wahlbring-at-crpgl.lu] Sent: Monday, August 06, 2001 10:49 AM To: Beauregard, Paul A.
Petra, we don't do much polymer work at my Lab, but I get a lot of them at materials ultramicrotomy workshops that I do. The key point is whether or not it is open-cell or closed-cell. If the former, it is pretty straightforward - use a low viscosity resin like Spurrs (but some ingredients are carcenogenic) or LR White (bit more viscous than Spurrs). Give it lots of time to infiltrate and, if possible, do so in a low vacuum to help encourage the air to exit the material.
If it is the latter - good luck! In 8 years of doing a hands-on workshop in the U.S. with a crack team of 7 experienced microtomists, we have been challenged with, and overcome, an amazing selection of students' samples for sectioning during the course of the week. We have only failed twice; once with a diamond-like carbon coating, but only because it was too thick (over a micron) and we had no way of reducing the thickness. The other was an innocent-looking, closed-cell polymer foam. Not only could we not embed it (quite understandably), we couldn't even fracture it after immersing in liquid nitrogen. (The outer layer would freeze but the interior remained as 'bouncy' as ever). If this is your case, I would find a FIB system and hope that it doesn't ion damage too much during ion sectioning.
Tom Malis Characterization Group Leader Materials Technology Laboratory Natural Resources Canada Ottawa, Canada
} From: Petra Wahlbring {petra.wahlbring-at-crpgl.lu} } Date: Mon, 06 Aug 2001 13:57:31 +0200 } To: microscopy-at-sparc5.microscopy.com } Subject: TEM: embedding media for PU/Polypyrrole-Foam? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear listers, } } the specimen on my desk is a polyurethane foam (about 100 pores/inch) } covered with polypyrrole. I would like to get a ultramicrotome cut of it } for TEM anlalysis. } Can anybody suggest a preparation method? Which resin should I try for } embedding and are there any other tips and tricks about this kind of } material? I want to look at an unstained specimen and I would like to try a } staining which will allow me to distinguish between the PU and Polypyrrol } (and the resin, of course). Will RuO4 or OsO4 work with this combination? } } Thanks for your input :) } } Petra } -------------------------------------------------------------- } Dr. Petra Wahlbring } Laboratoire d'Analyse des Materiaux (LAM) } Centre de Recherche Public - Gabriel Lippmann } 162a, av. de la Faiencerie, L-1511 Luxembourg } tel. +352-470261-503 } fax +352-470261-549 } e-mail: petra.wahlbring-at-crpgl.lu } http://www.crpgl.lu/lam
Hi All, Now this is something that I was warned out when I first pitched up here, but has only just happened over the past couple of weeks. We're getting random noise spikes turning up in the spectra on a couple of spectrometers. These do not occur at the same time on both spectrometers, nor in the same position if the spectrum is obtained again. Sometimes these don't occur at all. It got so bad that we had to call a halt to operations a few days ago, while we tried to suss out what was going on. The peaks were just a mess. After sorting out a few dodgy looking connections, the problem has all but disappeared. We still get a few intermittant spikes but just about ok enough to get going again. We have JEOL counting electronics JSM-XCU. These are connected in series and we find the problem only on 2 and 4. I wonder whether there is anybody out there who has had similar problems and found out what it was. We've swapped things around and it's not the pre-amps, and doesn't seem to be the modules themselves. Any help gratefully received, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (try your luck!) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
We have looked at polypyrrole by itself, so while I can't suggest anything concerning the sectioning of the PU foam, I can think of a possibility regarding the staining. Because both PU and PPy are nitrogenous materials, it is likely that RuO4 and OsO4 would be overkill (though if you are used to handling them, I wouldn't rule out trying). However, polypyrrole is easily doped with various counterions, one of which is iodide, which would be very electron dense, and should give good contrast. There ought to be something in the literature about how to do the doping.
On Mon, 6 Aug 2001, Petra Wahlbring wrote:
} the specimen on my desk is a polyurethane foam (about 100 pores/inch) } covered with polypyrrole ... I would like to try a staining which will } allow me to distinguish between the PU and Polypyrrol (and the resin, of } course). Will RuO4 or OsO4 work with this combination?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Hi, I´m a biologist working with yeasts from natural environments. I would like to know if anyone has or had experience in doing SEM and nuclear staining with yeasts. Or if you know someone who has.
I'm looking for a trimming block for an AO/Reichert Ultracut. The original part description is; catalog number 9-70-17-56 Trimming Block (for serial numbers greater than 365849).
I am interested in using aniline blue to localize callose in botanical material. Does anyone have a reference for the absorption and emission spectra of aniline blue under UV light?
We'll be installing a Tecnai F20 this fall. FEI recommends that the microscope be surrounded by curtains. It is not clear to me whether the curtains should be of a sound-attenuating material. Any advice from those who have already installed a Tecnai 20 (of any variety) would be appreciated.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 recook-at-anl.gov
Russel, We have installed one of the last CM 300's made and we used curtains to create a subroom around the scope for two reasons: 1) to reduce sound reverberations, 2) more importantly, as a focus for the air conditioning. The air coming into the room is outside of the curtains. The cold air falls to the floor and flows through a gap between the floor and curtains. The heat of the scope causes a gentle upward flow in its vicinity and is exhausted out a vent in a "chimney" built into the ceiling. Thus, we have cooling without much wind currents. Ciao for now, Ken
I have a used LKB 2168 TEM grid stainer needing adoption. The unit was in working order as of its last usage (approximately 4 yrs ago). I anticipate the system will only require a thorough cleaning prior to entering service at its new home. Any interested non-commercial parties should contact me off line for details.
Act quickly this is a limited time offer....the junk man cometh .
} John A. Robson } _____________________________________________________ } } Boehringer Ingelheim Pharmaceuticals, Inc. } Research and Development } 900 Ridgebury Road / P. O. Box 368 } Ridgefield, CT 06877-0368 } } phone: 203.798.5640 } fax: 203.798.5698 } email: jrobson-at-rdg.boehringer-ingelheim.com } } }
a few snapshots of Rachel & Natasha from last month are at http://drdcox.home.mindspring.com/family/ncc_rcc_july01.jpg
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
The Life Science Microscopy Facility at Purdue University seeks an electron microscopist for a university-wide, multi-user facility. The successful candidate will supervise the specimen preparation section of the microscopy facility. Duties include (1) prepare and photograph specimens for transmission and scanning electron microscopy, (2) instruct users in the proper use of electron and various light microscopes. (3) Participate in laboratory courses taught by faculty (4) Develop new procedures and techniques and furnish technical knowledge for interpretation of micrographs. (5) Maintain billing records for service activities. The candidate will collaborate with faculty and graduate students on research projects and in some cases will be considered a co-author.
Minimum academic requirements are a B.S. preferably in the life sciences. Salary is commensurate with experience. Purdue offers an outstanding benefit package.
Purdue is an affirmative action / equal opportunity employer. Women and minorities are encouraged to apply.
Applicants can send their CV to: (email acceptable) John J. Turek, Ph.D. Professor of Basic Medical Sciences Purdue University School of Veterinary Medicine W. Lafayette IN, 47907-1246 Phone: 765-494-5854 Fax: 765-494-0781 email:turekj-at-purdue.edu
I have a small (actually not so small), but interesting problem which I hope people out there in microscopy land may be able to help me with.
We have submitted two separate applications for replacement EM's. One application was for a FEG-SEM with cold stage and the other for a 120kv TEM with cryoholder. Both these configurations had been chosen after much consultation with the researchers who use our Unit. The applications were well supported by these researchers.
The response from our Equipment committee follows;
"Taking both applications into consideration, the committee would like to ask you to examine the feasibility of applying for a scanning transmission electron microscope (STEM), which the committee believe may better facilitate research. Can you please investigate whether the purchase of such an instrument would be a viable option for the University in enhancing its biological science research ?"
We in the Unit believe that only a FESEM and a cryo-TEM will meet all the needs expressed to us by researchers and that the purchase of a STEM will mean spending a significant amount of money for a backward step in 'real need' capability.
However I have to write a report justifying this belief.
Unfortunately I have no experience with STEM nor do I know a great deal about STEM. As I see it a STEM with cryo-capability may well be a possible option for our TEM researchers but my reason for not wishing to pursue the STEM option is simply because the trade off would be that a significant number of researchers at this University who have expressed a need for access to a cryo-capable high resolution SEM to study frozen hydrated 'large' samples will not have their needs meet. The purchase of a cryo-capable STEM will result in other compromises. For example, I understand that to have a cryo-capable STEM means we would have to compromise access to a back scattered electron detector. Due to design considerations a cryo-capable STEM can not have a back scattered electron detector fitted.
Any ideas to help me write my report will be gratefully received. Ideas to the contrary of those expressed above are also very very welcome.
Replies please to Allan Mitchell, allan.mitchell-at-stonebow.otago.ac.nz.
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
I am looking for a used dimpler to prepare TEM samples. Any information about it would be greatly appreciated.
Best regards,
Zhiping LUO Research Scientist Electron Microscopy Center Biological Sciences Building West Texas A&M University College Station, TX 77843-2257 Phone: (979) 845-1129 FAX: (979) 847-8933 E-mail: luo-at-emcnext2.tamu.edu
Check out 'The study of Plant Structure: Principles and Selected Methods' by O'Brien and McCully (1981), the bible of preparing plant tissue for microscopy. On pg 6.98, they give the 'peak absorption maximum for the dye-tissue complex' as ca. 370 nm, and the 'peak fluorescence emission' as 509 nm. I hope this helps.
Dear all, Does anybody have any good ideas about a suitable EDX standard for Y:Si?
I am working on yttrium silicates, and they tend to have fairly variable compositions (despite the fact that they are recorded as line compositions on the phase diagrams). Perhaps a well characterised, homogeneous glass containing Y?
I hope there are some good ideas out there?
Thanks in advance
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/ MPI-Stuttgart (mostly in German): http://www.mpi-stuttgart.mpg.de/
Our Astimex WDS / EDS standards for Y use Y3Al5O12 which is a synthetic. I suppose this is used because Astimex recognized years ago that this has better homogeneity than the Y in Si. Disclaimer: ProSciTech supplies these standards. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, August 10, 2001 12:11 AM, Ian MacLaren [SMTP:maclaren-at-hrem.mpi-stuttgart.mpg.de] wrote: } { { File: ATT00008.txt; charset = Windows-1252 } } } } } Dear all, } Does anybody have any good ideas about a suitable EDX standard for Y:Si? } } I am working on yttrium silicates, and they tend to have fairly variable } compositions (despite the fact that they are recorded as line compositions } on the phase diagrams). Perhaps a well characterised, homogeneous glass } containing Y? } } I hope there are some good ideas out there? } } Thanks in advance } } Ian MacLaren } Max Planck Institut fur Metallforschung, Seestra?e 92, } 70174 Stuttgart, Germany } Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk } Home page: http://members.tripod.co.uk/IanMacLaren/ } MPI-Stuttgart (mostly in German): http://www.mpi-stuttgart.mpg.de/
The Delaware Biotechnology Institute (DBI) at the University of Delaware has an immediate opening for a Research Associate I in its state-of-the-art BioImaging Center. The successful candidate should have a BS/MS in Biology or related field and a strong background (2+ years) in microscopy. The BioImaging Center includes an array of microscopy equipment including conventional fluorescence, confocal, multi-photon, atomic force, transmission and field emission scanning electron microscopes and their ancillary sample preparation equipment.
The primary responsibilities of the position include assisting the director of the BioImaging Center with general day-to-day operation of the multi-user facility as well as user training, equipment scheduling, and report generation. Experience in a multi-user facility and good familiarity with digital image processing and output, confocal microscopy and biological EM sample preparation are desirable. The individual must have strong communication and organizational skills, be motivated to learn new techniques, flexible, and have the ability to interact with a diverse group of research personnel. The highly variable nature of projects and collaborations as well as the excellent facilities, provides tremendous opportunity for professional growth.
The salary for the position will be commensurate with successful candidate qualifications. Applicants should send a cover letter, resume/CV, and list of 3 references to Professor Kirk J. Czymmek, 15 Innovation Way, Delaware Biotechnology Institute, University of Delaware, Newark, DE 19711. Email: kirk-at-udel.edu
Applications will be accepted until August 20, 2000 or until the position is filled.
The University of Delaware is an Equal Opportunity Employer, which encourages applications from Minority Group Members and Women.
We do have a used D500 Dimpler that just became available. This is not the same as our newer version D500i, but it is still an excellent product. If you have an interest, please contact me off-line.
Best regards-
David -- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
Zhiping Luo wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All, } } I am looking for a used dimpler to prepare TEM samples. Any information } about it would be greatly appreciated. } } Best regards, } } Zhiping LUO } Research Scientist } Electron Microscopy Center } Biological Sciences Building West } Texas A&M University } College Station, TX 77843-2257 } Phone: (979) 845-1129 } FAX: (979) 847-8933 } E-mail: luo-at-emcnext2.tamu.edu
The problem: to install a Nikon Coolpix 990 on a MEF-3 Reichert-Jung (metallurgical) light microscope.
We bought from Nikon the MCD lens adapter, which fits well the standard port for a CCD camera on the left-hand side of the microscope – actually a “T” connection on the “Rotoscope” screen support tube.
However:
(1) The field-of-view grabbed and displayed by the Coolpix is considerably smaller (just one quarter linear size) than the field-of-view of the Polaroid camera or that seen through the binoculars, for the same specimen and microscope setting. It is noteworthy that the image itself is distortion-free and of high quality.
(2) No in-focus image can be obtained when the microscope is set to magnification of x50 and below.
(3) The same problems appeared also when fitting a CCD camera through the same port.
(4) The problem can be solved by mounting the Coolpix instead of one of the two eyepices (occulars). Obviously, this is a bad solution, as the microscope is left with only one eyepiece.
I have seen an adaptor invented and marketed by Martin Microscope Company designed for the Cool Pix camera. It was quite impressive. I have no financial ties to the company above.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ARSEG -at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, August 10, 2001 at 18:21:39 ---------------------------------------------------------------------------
Email: ARSEG -at-aol.com Name: Rene Griffith
Organization: Union
Education: 9-12th Grade High School
Location: City, State, Country
Question: Hi my name is rene.I am a researcher at union highscool and i will be developing a slide to observe mycorrhizal relationships. So i would like to know what mountant you think i can use?
I'm not sure about your specific microscope model, but I suspect that Optem has an adapter which will fit it.
Optem makes a very nice adapter system for the 990. The top piece is PN 257015 which mates to PN 29-90-56. This then mates to a standard 1X C-mount on the trinoc port of the microscope. The lens inside the adapters provides the correct field of view to match that seen through the oculars.
Optem is in Fairport, NY USA and can be reached at info-at-optemintl.com or URL http://www.optemintl.com
gary g.
At 01:01 AM 8/11/2001, you wrote:
} Hi all, } } Connect Coolpix 990 to a light microscope } } Can anybody help? } } The problem: to install a Nikon Coolpix 990 on } a MEF-3 Reichert-Jung (metallurgical) light } microscope. } } We bought from Nikon the MCD lens adapter, } which fits well the standard port for a CCD } camera on the left-hand side of the microscope } actually a "T" connection on the "Rotoscope" } screen support tube. } } However: } } (1) The field-of-view grabbed and displayed by the } Coolpix is considerably smaller (just one quarter } linear size) than the field-of-view of the Polaroid } camera or that seen through the binoculars, for } the same specimen and microscope setting. It is } noteworthy that the image itself is distortion-free } and of high quality. } } (2) No in-focus image can be obtained when the } microscope is set to magnification of x50 and } below. } } (3) The same problems appeared also when fitting } a CCD camera through the same port. } } (4) The problem can be solved by mounting the } Coolpix instead of one of the two eyepices (occulars). } Obviously, this is a bad solution, as the microscope } is left with only one eyepiece. } } Is there a remedy? } } } Thanx, } } Dr. Uri Admon } } Beer-Sheva, Israel
I am wondering if anyone can help point me in the right direction regarding status of research, or better yet, ready to go software for computing 3D reconstruction of an SEM image.
My particular interest is defects on wafer surfaces. I have seen stereo pairs and some materials on reconstruction from stereo pairs, but not too impressed so far for my needs (not to say I have not seen some dandy stero pair SEM images).
I was thinking one could acquire multiple views (i.e., analogous to top, front, right side, rear, left side ) accomplished by rotation and tilt in the SEM. Then perhaps with a little help from the user to define common points in the various views, one to reconstruct the image. I am oversimplifying and no offense to the image experts, but the rest is math and graphics.
Anyone know of something like this or functionally equivalent ?
Regards, Ed Principe Defect & Thin Film Characterization Laboratory Applied Materials
Ed Principe wrote: ============================================================== I am wondering if anyone can help point me in the right direction regarding status of research, or better yet, ready to go software for computing 3D reconstruction of an SEM image.
My particular interest is defects on wafer surfaces. I have seen stereo pairs and some materials on reconstruction from stereo pairs, but not too impressed so far for my needs (not to say I have not seen some dandy stero pair SEM images).
I was thinking one could acquire multiple views (i.e., analogous to top, front, right side, rear, left side ) accomplished by rotation and tilt in the SEM. Then perhaps with a little help from the user to define common points in the various views, one to reconstruct the image. I am oversimplifying and no offense to the image experts, but the rest is math and graphics.
Anyone know of something like this or functionally equivalent ? =============================================================== Two such firms offering this kind of product are
1] Alicona (Germany) and their MeX 3D Reconstruction Software from SEM images www.alicona.com
and
2] Soft Imaging Systems (also Germany) http://www.soft-imaging.de/
There are some differences between the two products, including price. We have use both in our own laboratory and find them both quite acceptable.
Disclaimer: SPI Supplies will be distributing the Alicona product in the near future.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
In a message dated 8/12/01 1:43:03 PM, cgarber-at-2spi.com writes:
} My particular interest is defects on wafer surfaces. I have seen stereo } pairs and some materials on reconstruction from stereo pairs, but not too } impressed so far for my needs (not to say I have not seen some dandy stero } pair SEM images). } } I was thinking one could acquire multiple views (i.e., analogous to top, } front, right side, rear, left side ) accomplished by rotation and tilt } in } the SEM. Then perhaps with a little help from the user to define common } points in the various views, one to reconstruct the image. I am } oversimplifying and no offense to the image experts, but the rest is math } and graphics. } } Anyone know of something like this or functionally equivalent ?
Fovea Pro (http://reindeergraphics.com) includes this capability, as do several more costly packages. The Fovea routine functions as a plug-in within Photoshop. It uses cross-correlation to match points between the images, producing an image in which the horizontal displacement (parallax) is recorded as a grey scale value. This image can be measured (e.g., line profiles, etc.) or used to render perspective-corrected graphic images of the surface.
} Date: Sun, 12 Aug 2001 09:04:08 -0700 } To: Edward_Principe-at-amat.com } From: Gary Gaugler {gary-at-microtechnics.com} } Subject: Re: 3D SEM Reconstruction } } } Take a look at AutoQuant 3D reconstruction software. } I use it an I am an authorized dealer for AQI products. } } You can see more about their Autovisualize 3D at } } http://www.aqi.com } } gary gaugler } 916.791.8191 (Calif.) } } } At 10:21 PM 8/11/2001, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have a Reichert MeF-3 microscope and have mounted a Kodak Megaplus 1.4 on the right side, which is a port for ther macro camera. A coupling lens from Diagnostic Instruments was needed to fill the camera detector. We had mounted it on the left side but in re-arranging the room we moved it to the other side without difficulty.
Best Regards
Sam Purdy
Technical Center National Steel Corp. 1745 Fritz Dr. Trenton MI Voice:734-676-2682 FAX: 734-676-2682 spurdy-at-nationalsteel.com
-- Electron Microscopy position available. Multi-user; clinical/research facility. Call Valerie Sinor -at- 210-567-2607 for more information. UTHSCSA, Department of Pathology, San Antonio, Texas EQUAL OPPORTUNITY/AFFIRMATIVE ACTION EMPLOYER
-- Electron Microscopy position available. Multi-user; clinical/research facility. Call Valerie Sinor -at- 210-567-2607 for more information. UTHSCSA, Department of Pathology, San Antonio, Texas EQUAL OPPORTUNITY/AFFIRMATIVE ACTION EMPLOYER
Does anyone out there in cyber space have any experience in dealing with periodic acid methenamine silver staining of renal basement membranes at the ultrastructural level (that is thin sections). Any help would be gratefully appreciated
Many thanks
Terry
Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6009
Dear Richard, While an STEM may be a viable substitute for a TEM for some applications, it cannot do what an SEM can do. 1. STEM runs at 100 to 200 kV accelerating voltage for beam penetration, the opposite of the surface studies you want to do at 1 to 20 kV on a FESEM. 2. Sample size will be restricted to a standard TEM holder: 3 mm in diameter and 0.5 mm thick. Most SEM samples are 10 mm cubed or larger. 3. A modern STEM will cost about twice as much as a FESEM. The only dedicated STEM I saw in the Exhibits of the M&M 2001 last week was aimed at the semi-conductor testing market and there were no biological application mentioned. I have a TEM with scanning option, but the restrictions above also apply to it. The STEM mode is used for EDX testing. Anyone who thinks an STEM can substitute for an SEM has no knowledge of either technology. At 09:18 AM 8/9/01 +1200, you wrote: } } Greetings all. } } I have a small (actually not so small), but interesting problem which I } hope people out there in microscopy land may be able to help me with. } } We have submitted two separate applications for replacement EM's. One } application was for a FEG-SEM with cold stage and the other for a 120kv } TEM with cryoholder. Both these configurations had been chosen after much } consultation with the researchers who use our Unit. The applications were } well supported by these researchers. } } The response from our Equipment committee follows; } } "Taking both applications into consideration, the committee would like to } ask you to examine the feasibility of applying for a scanning transmission } electron microscope (STEM), which the committee believe may better } facilitate research. Can you please investigate whether the purchase of } such an instrument would be a viable option for the University in enhancing } its biological science research ?" } } We in the Unit believe that only a FESEM and a cryo-TEM will meet all the } needs expressed to us by researchers and that the purchase of a STEM will } mean spending a significant amount of money for a backward step in 'real } need' capability. } } However I have to write a report justifying this belief. } } Unfortunately I have no experience with STEM nor do I know a great deal } about STEM. As I see it a STEM with cryo-capability may well be a possible } option for our TEM researchers but my reason for not wishing to pursue the } STEM option is simply because the trade off would be that a significant } number of researchers at this University who have expressed a need for } access to a cryo-capable high resolution SEM to study frozen hydrated } 'large' samples will not have their needs meet. } The purchase of a cryo-capable STEM will result in other compromises. For } example, I understand that to have a cryo-capable STEM means we would have } to compromise access to a back scattered electron detector. Due to design } considerations a cryo-capable STEM can not have a back scattered electron } detector fitted. } } Any ideas to help me write my report will be gratefully received. Ideas } to the contrary of those expressed above are also very very welcome. } } Replies please to Allan Mitchell, allan.mitchell-at-stonebow.otago.ac.nz. } } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences, University of Otago } PO Box 913, Dunedin } NEW ZEALAND
Regards and good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Our group uses a CAMECA SU 30 SEM Probe.Recently it has developed a problem in CRT.The image starts shaking when it is continued to be on for some time.We thought it may starts heating up when is kept on so we put a cooling fan in it and also replaced the old heat sinks in the circuits with new ones. Still we could not solve the problem although it decreased to a little extent.
I do hope to get suggestions to overcome the problems.
Thanks in advance
Regards
Mr. Pankaj Kumar Patro PhD student IIT-Bombay Mumbai,India
Hello, Anyone know what non and artifactual causes are there for membrane blebbing or sloughing of parasite intected intestinal cells. We know about glut and over microwaving (insufficient lipid fix and overheating) anything else? thanks Mike D
We are going to purchase an electropolisher for our electron microscopy lab. We intend to use the polisher to prepare metallic TEM samples. Right now I have three candidates, Fishione Model 110 Twin-Jet Electropolisher, Struers TenuPol-5, and South Bay Model 550D. Can anyone with experiences using one of the polishers make comments to help us make decision?
Please contact me offline. My meail address: gongw-at-vsl.cua.edu
Dear Listers, I am having some trouble out of my SEM. Problem is in evacuation no led's are lighting up on the sequence panel. I can hear the rotary pump running, but I am not sure if it is actually working. So far I have checked the rotary pump oil level. Disassembled cleaned and reassembled the RP Vent valve. Checked the diffusion pump heater in and out of circuit. And of course the fuses. Working voltage is 100 volt AC and line voltage as checked is running at 108 volts. Switching to VAC on Checker Meter shows zero..........Any help would be appreciated Thank you in advance
p.s. VITALY FEINGOLD are you still out there I have lost your e-mail address
Robert Fowler Quality Assurance Technician TDK Components USA, Inc. 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.tdk.com
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We are going to purchase an electropolisher for our electron microscopy lab. We intend to use the polisher to prepare metallic TEM samples. Right now I have three candidates, Fishione Model 110 Twin-Jet Electropolisher, Struers TenuPol-5, and South Bay Model 550D. Can anyone with experiences using one of the polishers make comments to help us make decision?
Please contact me offline. My meail address: gongw-at-vsl.cua.edu
I would say the very crudest breakdown of STEM and TEM is: TEM can produce some of the highest signal/noise high resolution phase images (i.e., TEM is good for imaging). The extreme example is 'true' HRTEM. STEM is primarily valued for its spectroscopic analytical capability (not implying at all TEM does not provide analytical capability). The very latest/greatest HRTEM can match STEM image resolution. The STEM's most common imaging mode, high angle dark field, is primarily a Z-contrast image. Today's latest TEM/STEM systems can operate in STEM mode with a ~1.4A (300kV) or ~1.9A (200kV) and spot size with sufficient current to collect (P)EELS and EDS data in 5-20s per point (S/N issue). With a monochromoter, source correction, Cs hardware correction and the latest energy filter/energy loss spectrometer; you can approach meV energy resolution (should be coming "soon"). This is a change from earlier generations of TEM/STEM, where only a dedicated STEM in a research level facility could approach this spot resolution. The jist is; today or soon, you can really get the best of both worlds in a single commercial instrument. Obviously, I am a bit excited about the possibilities.
I think it really comes down to this: Do you need technology at your disposal that will allow you to examine the composition and chemistry of interfaces, cross-sections and surfaces at near atomic resolution by the methods accessible by TEM/STEM.....and you got money (~$2.5M for the hardware)? If yes, you need a STEM. Can you live with pictures and generic elemental EDS/WDS to get the information you want ? If yes, SEM-based information for you. SEM-Based STEM is somewhere in between, but much closer to SEM than STEM. You can approach some of the same image gains provided by STEM and also some of the enhanced spatial resolution EDS benefits by working with a TEM thin ( {2kA) sample on a SEM-based STEM system, but really not the same thing. But, you might have a practical niche.
I omitted a lot of caveats to the above statements, and there are usually multiple ways (analytical approaches) to achieve any given end, but that is the big picture. STEM is up there in terms of near atomic resolution chemical / elemental information.
You mentioned biological research. That niche in particular has a host of "pecadillos" and you might need to track down an expert or five when thinking about STEM. I have zero biological experience, so can offer no bio-specific help. But, you got a lot of current in a small spot. PEELS and especially GIF offer some time/spot advantages that I would find generically appealing if I were in a biological field, but I'm not. I bet there are people who know. How many people are using STEM/cryo in biological research? Why? I also can't comment of geometry and manufacturing limits for cryo STEM verses back-scatter detector.
Regarding SEM in general; well, that is a different type of information access. SEM yields images that when supplemented with various analyzers/detectors can provide topography info (standard detectors, immersion lens, multi-perspective detectors, stereo pair images, etc), "material contrast" info (i.e., BSE detector) and chemical information (primarily EDS and less common WDS/micro-probe), as well as some structural information (including BSED..back scatter electron diffraction). Peolple even stick more exotic and generally less robust analyzers (ie., electron flight tube/AES) on a SEM-based tool. I know I am forgetting a host of other relevant information. There are people who specialize in SEM for a career who know much much better.
SEM sample prep is less demanding, in general, but is also an art form and widely varied (biological could be the tougher). TEM sample prep is demanding, again being very general, and until very recently, was also very dependent on an individual's skill. With the advent of FIB-based "semi-automatic" TEM sample prep, things are getting interesting, if not easier (may not apply to bio applications, ....or could it?). But, there is still no replacement for a skilled operator and a sample prep expert. You can only analyze what you can prepare successfully.
Facilities costs will be considerably higher for a good TEM/STEM operation. The amount of work anticipated is always a factor when justifying capital. Just biological ? Anybody else to support the capital?
Regarding TEM/STEM, there is also a difference in an operator skilled at "imaging" (TEM) and an operator skilled at "analysis" (let's say STEM). The latter is more rare, but there is high value if you find one who fits your bill. I believe TEM/STEM is on the verge of some exciting new growth in terms of applications and general utility.
BIG Disclaimer: I do not represent or work for any SEM, TEM, or STEM manufacturer.
Much more than you likely wanted, but I hope it helps. You may have stumbled into a can of worms. But, I can sympathize with those "beneath" you who count on your educated decision and well-written report. Please appreciate the complexity of the decision matrix laid before you. It may pay significantly if you make the right decision and conversely, costly in terms of either opportunity or actual hardware cost if you make a poor decision.
Good Luck. I'd be intrigued to know the final decision and reasoning. Regards, Ed
Edward L. Principe, Ph.D. Member of Technical Staff Defect & Thin Film Characterization Laboratory Applied Materials
Mary Mager {mager-at-interchange.ubc.ca} on 08/14/2001 09:03:00 AM
To: Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} cc: Microscopy-at-sparc5.microscopy.com
Dear Richard, While an STEM may be a viable substitute for a TEM for some applications, it cannot do what an SEM can do. 1. STEM runs at 100 to 200 kV accelerating voltage for beam penetration, the opposite of the surface studies you want to do at 1 to 20 kV on a FESEM. 2. Sample size will be restricted to a standard TEM holder: 3 mm in diameter and 0.5 mm thick. Most SEM samples are 10 mm cubed or larger. 3. A modern STEM will cost about twice as much as a FESEM. The only dedicated STEM I saw in the Exhibits of the M&M 2001 last week was aimed at the semi-conductor testing market and there were no biological application mentioned. I have a TEM with scanning option, but the restrictions above also apply to it. The STEM mode is used for EDX testing. Anyone who thinks an STEM can substitute for an SEM has no knowledge of either technology. At 09:18 AM 8/9/01 +1200, you wrote: } } Greetings all. } } I have a small (actually not so small), but interesting problem which I } hope people out there in microscopy land may be able to help me with. } } We have submitted two separate applications for replacement EM's. One } application was for a FEG-SEM with cold stage and the other for a 120kv } TEM with cryoholder. Both these configurations had been chosen after much } consultation with the researchers who use our Unit. The applications were } well supported by these researchers. } } The response from our Equipment committee follows; } } "Taking both applications into consideration, the committee would like to } ask you to examine the feasibility of applying for a scanning transmission } electron microscope (STEM), which the committee believe may better } facilitate research. Can you please investigate whether the purchase of } such an instrument would be a viable option for the University in enhancing } its biological science research ?" } } We in the Unit believe that only a FESEM and a cryo-TEM will meet all the } needs expressed to us by researchers and that the purchase of a STEM will } mean spending a significant amount of money for a backward step in 'real } need' capability. } } However I have to write a report justifying this belief. } } Unfortunately I have no experience with STEM nor do I know a great deal } about STEM. As I see it a STEM with cryo-capability may well be a possible } option for our TEM researchers but my reason for not wishing to pursue the } STEM option is simply because the trade off would be that a significant } number of researchers at this University who have expressed a need for } access to a cryo-capable high resolution SEM to study frozen hydrated } 'large' samples will not have their needs meet. } The purchase of a cryo-capable STEM will result in other compromises. For } example, I understand that to have a cryo-capable STEM means we would have } to compromise access to a back scattered electron detector. Due to design } considerations a cryo-capable STEM can not have a back scattered electron } detector fitted. } } Any ideas to help me write my report will be gratefully received. Ideas } to the contrary of those expressed above are also very very welcome. } } Replies please to Allan Mitchell, allan.mitchell-at-stonebow.otago.ac.nz. } } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences, University of Otago } PO Box 913, Dunedin } NEW ZEALAND
Regards and good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Any experience with plastic coverslipping machines? We are having problems with poor resolution from our two year old system. Looking at a glass cover slipped slide the microscope resolution is great, changing to one of the plastic ones and it's not so good... It could be processing or the plastic, the glass cover slipped slides are done by a different facility.
Does anyone out there know of a good source, text or website, that deals with the issue of contaminants in a semiconductor manufacturing environment? I'm putting together a lecture on the subject but I don't want to generate SEM's, optical images etc. if this is available readily. Principally I'm interested in human spittle, airborne contaminates such as skin, hair, cloth fibers, etc. or anything that may be generated by human interaction with a semiconductor device.
Mike: Back when biological SEM was new, there was a lot of excitement over the use of cell surface blebbing, etc to differentiate between T and B cells. Well, the upshot of things (and I forget the references) was that the heating of the sample during critical point drying was causing the blebbing, ruffles, etc. And this was after both glutaraldehyde and osmium fixation. So, my guess is that that the blebbing is caused by heating. Membranes are still very mobile (the lipid phases, that is) following fixation, and driving the temp up to 50C or higher will probably give you some artifacts. Question is, can you live with those artifacts? I would also recommend that you check the Ted Pella, Inc website (http://www.tedpella.com) for protocols using the microwave at lower temps. There were a number of excellent papers at last week's Microscopy & Microanalysis meeting using these techniques. Steve Slap at Hacker might also be a good resource for you here. Roger On Tue, 14 Aug 2001 13:02:31 -0400, Mike Delannoy wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hello, | Anyone know what non and artifactual causes are there for membrane | blebbing or sloughing of parasite intected intestinal cells. We know | about glut and over microwaving | (insufficient lipid fix and overheating) anything else? | thanks | Mike D | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I've received at least a dozen emails sent by this virus over the last few weeks, so watch out for it. I highly recommend viewing messages on the mail server BEFORE downloading your mail (use message dispatcher in The Bat mail client). Then you can delete the message directly from the server.
} You may receive a message sent by SirCam virus. The body of the } message is randomly chosen, although the first and last lines are } always "Hi! How are you?" and "See you later. Thanks" in the English } version of the message and "Hola como estas?" and "Nos vemos pronto, } gracias" in the Spanish version. The message always have a file } attachment with double extension, which is also randomly choosen, } e.g .doc.bat, .doc.lnk, .xls.bat. Altough The Bat! will warn you } before opening such files or blocks them completely, do not try to } open them!
You will not get a copy of the SirCam virus by way of the Microscopy Listserver. My software has been blocking it fine (for now), however, the server is slow now because of the number of attacks I'm getting. Each attempt as Tony points out has an attached file which is starting to cause headaches for me as the server drives are filling and emptying constantly. When the drives fill we hit a bottle neck and the server grinds to a halt until the junk mail and it's attachments are cleared.
Nestor Your Friendly Neighborhood SysOp.
PS, I wish I was getting only a dozen attempt over a few week period. I've been getting over 500/day as of this weekend. Sigh..the joys of being a SysOp...
It sure was nice to see so many of you at M&M 2001 in Long Beach last week!
I generally only look at 0.5 to 2.0 micrometer epoxy or acrylic sections in the light microscope to see where in the block I am before taking ultrathin sections for TEM. I stain the thick sections with toluidine blue, glance at them and then throw them away. However, now I want to keep some of them and get good photomicrographs as well. In the past when I coverslipped them and took (35 mm film) pictures, they looked pretty bad. I've heard of various fixes, such as using thicker coverslips.
Now I'd like to hear from someone doing this routinely, with good results (if possible!). What kind of mounting media? What kind of coverslips? Why? What goes on with the light as it passes through the resin?
What kind of optics? Phase? Hoffman? DIC? Plain vanilla BF?
Any clues will be appreciated!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The problem is I have a surface mount ceramic chip capacitor exhibiting a degradation in internal resistance but not so severe that it is readily visible using light microscope and my SEM is down for repairs. Probable cause is a microcrack and I need a suitable stain or dye penetrant to be able to detect. If necessary I can penetrate using vacuum. I would appreciate your resources and input on this problem. If there is a more suitable forum....I do remember some mention of EDFAS which I plan to join (Don't have time right now) please forward
Thanking you in advance
Robert Fowler Quality Assurance Technician TDK Components USA, Inc. 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.tdk.com
THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL INFORMATION.
If you are not the intended recipient, be advised that any use, dissemination, distribution or duplication of this transmission is strictly prohibited. If you received this transmission in error, please notify the sender immediately by electronic reply to this transmission or by phone (847-803-6100). Thank you.
I experienced a similar problem a few years back with a JEOL 6300. As ridiculous as it may sound, it turned out to be the nitrogen tank. It simply ran out of gas. So, if you have a N2 tank attached to your scope, check it. You may be in luck and just need to exchange the tank. Good Luck!
Jackie Garfield Electron Microscopist LifeCell Corportation One Millennium Way Branchburg, NJ 08876
We have a Wentworth LCS-2150 prober which is fitted with a Hoya L212-62A laser head and a Hoya L-210-P2 power supply. We do not have the user manuals for this system and cannot make it work. The laser fires but nothing happens to the chips under the Mituyo objectives. Power supply is set at 12KV. Simmer is completed but no pulse at the target.
Does anyone have this Hoya system and associated manuals? I would greatly appreciate a copy of any documentation. Reproduction and shipping costs will be gladly paid.
As far as I understand this instrument is intended to be for bio/medical applications. In my experience, STEM doesn't have any serious advantage over TEM for those applications (I don't have too much experience with bio applications, though).
But by all means, if you have enough money, get TEM/STEM not just TEM. Most of the popular TEM manufacturers give you STEM as an option. I wouldn't go with a dedicated STEM in your case. Here's what STEM migh give you: 1. You might get higher contrast in thicker samples than with TEM. 2. Z-contrast might be very helpful. 3. You can image with low dose and get decent images. 4. Extended analytical capabilities (you can conveniently collect all kinds of signals but you need to have all kinds of detectors for that). Since you are going to get a non FEG (I suppose) your resolution in STEM will probably be not as good as in TEM.
The reason why your committee came up with the STEM suggestion might be because, in fact, you can use a STEM in SEM mode. Just lower the voltage down to 20kV or so and if you have an SE or BSE detector- you have an SEM. So,
5. TEM/STEM can be put in SEM mode.
Hope this helps,
Max __________________________________ Max Sidorov, Ph.D. Advanced Micro Devices Sunnyvale, CA
-----Original Message----- } From: Richard Easingwood [mailto:richard.easingwood-at-stonebow.otago.ac.nz] Sent: Wednesday, August 08, 2001 2:18 PM To: EM Listserver - messages Cc: allan.mitchell-at-galadriel.otago.ac.nz; bronwyn.smaill-at-galadriel.otago.ac.nz
Greetings all.
I have a small (actually not so small), but interesting problem which I hope people out there in microscopy land may be able to help me with.
We have submitted two separate applications for replacement EM's. One application was for a FEG-SEM with cold stage and the other for a 120kv TEM with cryoholder. Both these configurations had been chosen after much consultation with the researchers who use our Unit. The applications were well supported by these researchers.
The response from our Equipment committee follows;
"Taking both applications into consideration, the committee would like to ask you to examine the feasibility of applying for a scanning transmission electron microscope (STEM), which the committee believe may better facilitate research. Can you please investigate whether the purchase of such an instrument would be a viable option for the University in enhancing its biological science research ?"
We in the Unit believe that only a FESEM and a cryo-TEM will meet all the needs expressed to us by researchers and that the purchase of a STEM will mean spending a significant amount of money for a backward step in 'real need' capability.
However I have to write a report justifying this belief.
Unfortunately I have no experience with STEM nor do I know a great deal about STEM. As I see it a STEM with cryo-capability may well be a possible option for our TEM researchers but my reason for not wishing to pursue the STEM option is simply because the trade off would be that a significant number of researchers at this University who have expressed a need for access to a cryo-capable high resolution SEM to study frozen hydrated 'large' samples will not have their needs meet. The purchase of a cryo-capable STEM will result in other compromises. For example, I understand that to have a cryo-capable STEM means we would have to compromise access to a back scattered electron detector. Due to design considerations a cryo-capable STEM can not have a back scattered electron detector fitted.
Any ideas to help me write my report will be gratefully received. Ideas to the contrary of those expressed above are also very very welcome.
Replies please to Allan Mitchell, allan.mitchell-at-stonebow.otago.ac.nz.
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
Recently I've been trying to sputter coat nickel onto biological samples for x-ray microanalysis (EDS) in a SEM but have not been able to get any coating under a variety of conditions onto a filter paper test sample. Gold works fine.
Short digression: I like to use nickel because, compared to evaporated carbon, it prevents charging more effectively, the K & L series nickel peaks are at very low and high energies respectively, so the biological peaks of interest are not overlapped, and you get better pictures of the analyzed area with a metal coat on it. I used to coat from nickel wire in a vacuum evaporator, but now have a sputter coater available on an SEM mounted cryo-prep unit, so also want to coat Ni onto frozen hydrated biological samples for EDS.
My unit is a D.C magnetron type. After reviewing the great discussions of sputter coating that took place in this forum in January, 2001, I have a hunch that i just don't have enough voltage in this unit to blast nickel atoms off the target.
Any suggestions on what to try next?
Also, maybe I should revisit carbon coatings - is it possible to sputter carbon without a special unit?
Thanks for any suggestions you might have,
Gib
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} Hi, All- } } It sure was nice to see so many of you at M&M 2001 in Long Beach last } week! } } I generally only look at 0.5 to 2.0 micrometer epoxy or acrylic sections } in the light microscope to see where in the block I am before taking } ultrathin sections for TEM. I stain the thick sections with toluidine } blue, glance at them and then throw them away. However, now I want to keep } some of them and get good photomicrographs as well. In the past when I } coverslipped them and took (35 mm film) pictures, they looked pretty } bad. I've heard of various fixes, such as using thicker coverslips. } } Now I'd like to hear from someone doing this routinely, with good } results (if possible!). What kind of mounting media? What kind of } coverslips? Why? What goes on with the light as it passes through the } resin? } } What kind of optics? Phase? Hoffman? DIC? Plain vanilla BF? } } Any clues will be appreciated! } } Aloha, } Tina
Tina,
I use with full sucess semithin sections (embedded in Spurr resin) mounted in Canada Balsam. However, it is important to remove the resin with a saturated solution of NaOH in ethanol, during 5 minutes (for 0.35 µm) before the staining procedures (Toluidine Blue or another technique). The photomicrographs taked in DIC or BF microscopies are perfect for me. I hope to have help you.
Dr. Rinaldo Pires dos Santos Dept. of Botany - UFRGS Rua Márcio Dias, 25 / apt. 305 Bairro Nonoai 90830-360 - Porto Alegre - RS Brazil E-mail: rinaldop-at-uol.com.br
Hi All: For several years I have struggled with finding the right encapsulant to encapsulate irregular shaped samples, powders, etcetera that will stand up well to heating, mechanical thinning, polishing and ion milling (most bow when heated and have poor adhesion). Does anyone out there(in Listserver Land) have any recommendations of an encapsulant that they use that has these type of characteristics? I would prefer users experiences, rather than sales pitches...
While I am on the subject...does anyone have recommendations of a protocol/procedure to process cross-sectional TEM specimens at room temperature or pretty much near room temperature? eg room temperature curing adhesives (to glue the samples together) and waxes or adhesives to temporarily attach the sample to the polishing puck (I have used Crazy Glue for this purpose, but it is not always reliable) to make cross-sections of let's say a temperature sensitive device? Materials science people will know what I am talking about (I hope). Thanks for your help, Regards, Michael Coviello Lab Manager UT Arlington
P.S. Thanks Nestor for a great job and an extremely valuable resource
We have an "old" Gatan Accuview 789 (based on Kodak Megaplus 1.4 or 1.4i, I'm not sure which) mounted on our TEM side (35mm?) port. We currently use it with a Mac. Sigh. We now want to have the capability to interface it to a PC. We do fairly simple imaging and limited image analysis (basic stuff). Can anyone tell me what we need? Easiest and/or cheapest route preferred. (Private replies to tcgruber1-at-excite.com are also welcome.)
TIA,
Tyler C. Gruber, Physicist III Physics/Microscopy Laboratory Columbian Chemicals Company
From root Fri Aug 17 08:46:04 2001 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA00884 for dist-Microscopy; Fri, 17 Aug 2001 08:15:45 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA00881 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 17 Aug 2001 08:15:14 -0500 (CDT) Received: from epostoffice.uta.edu (epostoffice.uta.edu [129.107.1.1]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA00874 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 17 Aug 2001 08:15:03 -0500 (CDT) Received: from mae.uta.edu (coviello.uta.edu [129.107.2.175]) by epostoffice.uta.edu (8.11.5/8.11.5) with ESMTP id f7HDAJQ24737 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 17 Aug 2001 08:10:19 -0500 (CDT) Message-ID: {3B7C6EF8.E956E91E-at-mae.uta.edu}
Hi All: For several years I have struggled with finding the right encapsulant to encapsulate irregular shaped samples, powders, etcetera that will stand up well to heating, mechanical thinning, polishing and ion milling (most bow when heated and have poor adhesion). Does anyone out there(in Listserver Land) have any recommendations of an encapsulant that they use that has these type of characteristics? I would prefer users experiences, rather than sales pitches...
While I am on the subject...does anyone have recommendations of a protocol/procedure to process cross-sectional TEM specimens at room temperature or pretty much near room temperature? eg room temperature curing adhesives (to glue the samples together) and waxes or adhesives to temporarily attach the sample to the polishing puck (I have used Crazy Glue for this purpose, but it is not always reliable) to make cross-sections of let's say a temperature sensitive device? Materials science people will know what I am talking about (I hope). Thanks for your help, Regards, Michael Coviello Lab Manager UT Arlington
P.S. Thanks Nestor for a great job and an extremely valuable resource
From root Fri Aug 17 08:46:04 2001 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA00995 for dist-Microscopy; Fri, 17 Aug 2001 08:22:16 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA00987 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 17 Aug 2001 08:21:45 -0500 (CDT) Received: from usatl1005.imagion.com ([216.52.197.250]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA00980 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 17 Aug 2001 08:21:33 -0500 (CDT) Received: from ccc-sysnt5.ccc.phelpsd.com ([198.176.209.2] unverified) by usatl1005.imagion.com with Microsoft SMTPSVC(5.0.2195.3779); Fri, 17 Aug 2001 08:54:02 -0400 Received: by ccc-sysnt5.ccc.phelpsd.com with Internet Mail Service (5.5.2650.21) id {QQ4CRSGB} ; Fri, 17 Aug 2001 09:18:38 -0400 Message-ID: {7F7A914EF10DD311AB0B00105A26EB7704FE90AD-at-ccc-sysnt5.ccc.phelpsd.com}
Hi Folks,
First-timer here.
We have an "old" Gatan Accuview 789 (based on Kodak Megaplus 1.4 or 1.4i, I'm not sure which) mounted on our TEM side (35mm?) port. We currently use it with a Mac. Sigh. We now want to have the capability to interface it to a PC. We do fairly simple imaging and limited image analysis (basic stuff). Can anyone tell me what we need? Easiest and/or cheapest route preferred. (Private replies to tcgruber1-at-excite.com are also welcome.)
TIA,
Tyler C. Gruber, Physicist III Physics/Microscopy Laboratory Columbian Chemicals Company
An older article by Bob Lowry, et al., at Harris (now Intersil) has images, elemental composition data, and interesting discussion of human contaminants on IC's.
"Analysis of Human Contaminants Pinpoint Sources of IC Defects" Lowry, R.K., Linn, J.H., Grove, G.M., and Vicroy, C.A. Semiconductor International, v 10 n 8 July 1987 p 73-77
Jack D. Henderson Naval Surface Warfare Center Crane Division
} -----Original Message----- } From: Peter Tomic [mailto:PTomic-at-anadigics.com] } Sent: Wednesday, August 15, 2001 6:56 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Semiconductor Contaminants } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListser} ver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } Does anyone out there know of a good source, text or website, } that deals } with the issue of contaminants in a semiconductor } manufacturing environment? } I'm putting together a lecture on the subject but I don't } want to generate } SEM's, optical images etc. if this is available readily. } Principally I'm } interested in human spittle, airborne contaminates such as } skin, hair, cloth } fibers, etc. or anything that may be generated by human } interaction with a } semiconductor device. } } Regards, } Peter Tomic } Anadigics, Inc. }
Many thanks to everyone who visited the RMC-Boeckeler Instruments booth and special thanks to those of you who took the time to fill out our survey and enter the drawing for a digital camera.
I am pleased to announce that the lucky winner is Mr. Jon Charlesworth at Mayo Clinic. Good luck Jon and hope you have lots of fun with your new "toy".
Sorry that you couldn't all be winners but better luck next time. See you in Quebec and be sure to visit our booth to try for next year's drawing.
Dave Roberts Director- RMC EM Products Boeckeler Instruments, Inc. 4650 S. Butterfield Drive Tucson, AZ 85714 Tel: (520) 745-0001 Fax: (520) 745-0004 website: www.rmcproducts.com
{!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.0 Transitional//EN"} Many = thanks to=20 everyone who visited the RMC-Boeckeler Instruments booth and special = thanks to=20 those of you who took the time to fill out our survey and enter the = drawing for=20 a digital camera.
I am = pleased to=20 announce that the lucky winner is Mr. Jon Charlesworth at Mayo = Clinic. =20 Good luck Jon and hope you have lots of fun with your new=20 "toy".
Sorry = that you=20 couldn't all be winners but better luck next time. See = you in Quebec=20 and be sure to visit our booth to try for next year's=20 drawing.
Dave=20 Roberts Director- RMC EM=20 Products Boeckeler=20 Instruments, Inc. 4650 = S. Butterfield=20 Drive Tucson, AZ=20 85714 Tel: = (520) 745-0001=20 Fax: (520) 745-0004 website: {http://www.rmcproducts.com"} www.rmcproducts.com=
To all you that asked, no it's not a problem with your Email account or your subscription.
The TelCo ( AT&T ) connnection to my router was down for nearly 24 hours.
The network simply failed. Somewhere between Chicago and here. For the moment all appears to be working but they are still doing testing. Probably some small animal chewing on buried cable or something equally silly, but hard to find.
We have a Wentworth LCS-2150 prober which is fitted with a Hoya L212-62A laser head and a Hoya L-210-P2 power supply. We do not have the user manuals for this system and cannot make it work. The laser fires but nothing happens to the chips under the Mituyo objectives. Power supply is set at 12KV. Simmer is completed but no pulse at the target.
Does anyone have this Hoya system and associated manuals? I would greatly appreciate a copy of any documentation. Reproduction and shipping costs will be gladly paid.
This is a survey that was passed out at the Facility Managers Meeting at this year's M&M meeting. We've only gotten 6 responses so far, and many more are needed for valid results (although obviously only one answer per facility). Please take a couple of minutes to fill this form in, and email back to me. Thank you!
Phil
MICROSCOPY FACILITY SUPPORT
This survey will help compile the nature and amount of support provided to microscopy facilities by universities, colleges, and research institutes. We would like to determine how much facility support is provided by the institution, and how much must be generated by user fees or other sources, on a per cent basis.
If we get enough responses to have valid data, we will post the results on the microscopy listserver and send them to the MSA bulletin. Names of institutions will not be included in the reported results.
Institution: Location: Nature of institution (Academic, Commercial, private research, etc.)
Type of Facility (institution core, satellite, departmental, etc):
(Place an 'X' by the appropriate answer) Predominately: Biological Materials Both
User base: Service Multi-user Both
Type of microscopes: SEM FESEM ESEM or LV TEM FETEM Major optical (Specify) Scanning Probe (specify) Other (specify)
Approximately what *per cent* of the funds for each of the following is provided by A) your institution, B) user fees, or C) grants
Salaries: A) B) C)
Instrument Maintenance (Service contracts, instrument insurance, etc): A) B) C)
Replacement of existing equipment: A) B) C)
Purchase of new equipment (not replacement for but addition to existing equipment): A) B) C)
Supplies: A) B) C)
Other operating expenses: A) B) C)
Are there other microscope facilities at your institution? if so, please indicate approximate number having electron microscopes. Ideally we would like a form filled out for each major facility (multiple instruments) at your institution if there are more than one.
Please include additional information such as novel funding ideas for your facility or for others at your institution.
Please email your responses to Philip Oshel peoshel-at-facstaff.wisc.edu -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
In about three weeks time or so, I'll be writing equipment grant appilcations. Our lab needs, as a matter of necessity, an analytical SEM. The one we have now is relatively old (Philips SEM515) and has no EDS facilities. It hinders most of our work since we have to go elsewhere and pay for any work involving EDS. Our attempt last year to get a EDS system grant for it failed. So, we are making a completely new equipment grant this time.
I am wondering if there are metallurgical microscopists in the house to give me some insights on which direction to go. What must we be looking for in a good analytical SEM? Who makes what?
Many thanks in advance.
Ike Oguocha Department of Mechanical Engineering University of Saskatchewan Saskatoon, Canada
____________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk or your free -at-yahoo.ie address at http://mail.yahoo.ie
Dear Listers, I have a student who's had little success attaching and then retaining neurons and PC12 cells to coverslips (glass, Thermanox and aclar) in an attempt to use a pre-embedd labeling procedure. She has coated glass with collagen for the PC12 culture. Any suggestions will be appreciated. Rosemary -- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
At 3:53 PM -0500 8/8/01, John J. Turek wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a question from a client that is beyond my area of expertise, so I turn to you for help.
He would like to look at the layer of ink deposited on paper by the traditional (Guttenberg type) printing press in order to help him standardize his printing process. I haven't a clue as to how to approach this. His idea was to look in SEM since his understanding is that the ink layer is, on average, 4 micrometers thick , but my feeling is that the ink will have absorbed into the paper and not present any topography.
Any ideas? I have TEM & SEM at my disposal, as well as CLSM and widefield LM.
Thanks in advance, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Purdue University seeks an Electron Microscope Specialist to provide service and support for the scanning and transmission electron microscopes, and related equipment, managed by the Purdue Electron Microscopy Consortium. Serving as a primary point-of-contact for all electron microscopy related concerns, questions and problems. He/she will manage the day-to-day maintenance of our equipment. He/she will also interface with manufacturers, provide training to users, train and supervise interns, and assist in the evaluation of proposed and existing equipment.
The successful candidate will possess at least an associates degree (or equivalent) in science, engineering or technology, suitable training in electronics and vacuum systems, and significant experience in the maintenance of electron microscopes and associated equipment.
This position will remain open until filled. For full consideration, applications should be received by October 31, 2001.
The Purdue Electron Microscopy Consortium oversees and co-ordinates the operation of nearly 30 electron microscopes (organized in four clusters) that support the educational and research missions of the university.
Purdue offers competitive salaries and an outstanding benefit package. Located in West Lafayette, Indiana, it provides a very attractive environment in which to live and work.
Applicants should send a letter of interest and a detailed CV to:
Prof. Alex King Director, Purdue Electron Microscopy Consortium School of Materials Engineering Purdue University West Lafayette IN 47907-1289
alexking-at-ecn.purdue.edu
Purdue is an affirmative action / equal opportunity employer. Women and minorities are encouraged to apply.
I recall a couple postings several months ago pertaining to old CRTs that were in need of repair. There were a couple responses with names of someone who repairs these old CRTs. I swore I printed those off but am unable to find it. If anyone has that info I would appreciate hearing from you. Thanks.
Joel McClintock EM Specialist U of Kentucky 859-257-1242
Dear Listers, } } I'm considering buying a used Sputter Coater originally marketed by Earnest } Fullam (I'm on a real shoestring budget). A web search doesn't turn up any } results for an extant company of that name. } } Anyone know whether the company still exists, and/or whether replacement } parts are available for such a unit? Thanks for any info. } } Cheers, } } Paul Grover :o)
I have a question from a client that is beyond my area of expertise, so I turn to you for help.
He would like to look at the layer of ink deposited on paper by the traditional (Guttenberg type) printing press in order to help him standardize his printing process. I haven't a clue as to how to approach this. His idea was to look in SEM since his understanding is that the ink layer is, on average, 4 micrometers thick , but my feeling is that the ink will have absorbed into the paper and not present any topography.
Any ideas? I have TEM & SEM at my disposal, as well as CLSM and widefield LM.
Dear Lee, What is the composition of the ink? If it has element(s) not present in the paper, perhaps you could use XMA or EELS to define the regions of the paper where the ink is. Cutting cross-sections perpendicular to the plane of the paper may give you enough spatial resolution, or you could try to prepare individual fibers if there is a technique to do this without redistributing the ink. This last method would give info about how far the ink gets into each fiber, but you would then have to find out how the fibers are oriented in the paper to get the ink distribution as a function of distance from the surface. This could be a difficult problem to unravel. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Thank you for taking the pain to go through my mail. My name is CHIEF MRS. MARYAM ABACHA. I would first like to apologise for this kind of introduction into your mailbox, but the situation I am now did not afford me any better way. It is with a heart full of hope that I write to seek your help in the context below. I am the wife of the former Nigerian Head of State late General Sani Abacha whose sudden death occurred on the 8th of June 1998. Having gotten your contact from the family's Library I have no doubt about your capability and goodwill to assist me in receiving into your custody (for safety). The sum of TWENTY ONE MILLION, SEVEN HUNDRED THOUSAND UNITED STATES DOLLARS (US$21.7 MILLION) WILLED and deposited in my favour by my late husband. The money is currently kept in a Trust deposited Account with one of the private Security Companies. As it is legally required, the Administration of my late husband is under the authority of the family's lawyer named (Mark Mercyke). However, the new Democratic Government has an assumption of office set up a panel of inquiry to probe the financial activities of my late husband (Former Head of State) with a decision to freeze all his assets respectively. The investigative team has submitted their report. Presently, some Bank Accounts and valuable assets have been frozen and seized. Fortunately, our family lawyer had secretly advised that the WILLED money be urgently moved into an unknown overseas Bank account of a trusted person or company without delay for security reasons. The Government had earlier placed an embargo on Abacha's family members from traveling outside the Country. My son (Mohammed Abacha) is in prison custody being tried for a crime he is innocent of . The situation has been so terrible that we are virtually. I expect you to be trustworthy and kind enough to respond to this call (S.O.S) to save me and my children from a hopeless future. I hereby agree with the Lawyer to compensate you with 25% of the total sum (US$21.7Million) for your sincere and candid effort when the money is finally received into your Bank Account. We have also set aside 5% of the total sum for settling any expenses you and I might incur during the process of the transfer of this money. That is to say, any amount you spend e.g. Telephone Bills, Security Charges etc. must be refunded to you before the sharing of the money and the same thing to us.The lawyer and the Security Company have already perfected arrangement with the manners to effect complete dislodgement of this money within 7 days of receipt of your response. They have equally guaranteed 100% RISK FREE and smooth transfer. Please, send the following information to the Lawyer immediately. (1) YOUR FULL NAME (2) YOUR ADDRESS (3) YOUR PHONE AND FAX NUMBERS. All CORRESPONDENCE/INQUIRY MUST BE DIRECTED TO THE LAWYER VIA THE E- MAIL ADDRESS:markmercyke-at-lawyer.com, as we count on your trustworthiness and confidentiality. Thanks for your kind attention. I look forward to your quick response. With Best Regards. Chief Mrs Maryam Abacha.
http://www.myplace.com/ - find what you want -at-MyPlace.com!
Thank you to all who have spotted my bad spellling (I plead innocence, as the advertisement had the spelling 'Earnest Fullam'). I'm glad so many are on vacation so that they don't see the egg on my face.
} Hello Netters, } } I have a question from a client that is beyond my area of expertise, } so I turn to you for help. } } He would like to look at the layer of ink deposited on paper by the } traditional (Guttenberg type) printing press in order to help him } standardize his printing process. I haven't a clue as to how to } approach this. His idea was to look in SEM since his understanding } is that the ink layer is, on average, 4 micrometers thick , but my } feeling is that the ink will have absorbed into the paper and not } present any topography. } } Any ideas? I have TEM & SEM at my disposal, as well as CLSM and widefield LM. } } Thanks in advance, } Lee
Depending on the coating on the paper and the composition of the ink the ink will be to a mostly deposited on the surface of the paper. Actually very little will be absorbed into the paper or you would have bleed through like that you see when you use Magic Markers or regular paper. It would also cause the type to bleed and become unsharp like you see when you use the wrong type of paper in an ink jet printer.
Printers ink is a compound of pigments, oils and dryers that that forms a polymer much like oil paint on paper. Very little of the drying it due to evaporation of solvents or the absorption of solvents by the paper. The ink is put in place and left to dry by transfer from the type to paper by very firm pressure.
Printing is just one of many things I learned to do in my life that technology has made obsolete.
Your customer knows what he is talking about.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
----- Original Message ----- } From: {secretmail-at-myplace.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, August 17, 2001 6:28 PM
Try this URL for their home page:
http://www.fullam.com/
This one for their sputter coater:
http://www.fullam.com/Effacoat.htm#18930
gary g.
At 03:23 PM 8/17/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
1. Weigh a piece of paper in an analytical balance. 2. Print on it and let it dry. 3. Weigh it again. 4. Use your flatbed scanner to scan the image. 5. Save as a black and white image. 6. Use an image processing program to tell what percent of the surface is black, ie. inked. (Many scanners come with software that allows this type of simple image processing.) 7. Do the math.
If you need to know what your error is do a number of these measurements.
Your error is probably far less than with the TEM.
We use a solution of 5 % gelatin in distilled water to attach cells to glass surface. It has to be diluted at 70 ºC on a magnetic shaker. Then the glasses are dept and dryed. It works.
Albert Cardona University of Barcelona
__________________________________________________ Do You Yahoo!? Make international calls for as low as $.04/minute with Yahoo! Messenger http://phonecard.yahoo.com/
Fixation: paraformaldehid 4% (merck) pepsin (sigma) 0,4 % in HCl 0,1 N pH 5 milk primary antibody biotinilated secondary antibody (sigma) avidin + biotin-peroxidase complex (you can get both apart in Sigma too) DAB revealing solution: DAB 0,3 % in PBS niquel amonium sulfate 0,2 % water peroxide 0,03 % -in PBS.
There can be found recipes on the net. Just write the name of any of the components on google or yahoo.
Albert Cardona University of Barcelona
__________________________________________________ Do You Yahoo!? Make international calls for as low as $.04/minute with Yahoo! Messenger http://phonecard.yahoo.com/
These were from my HP LaserJet 4 (laserpaper) and from page 61 of the Volume 7, Supplement 1 issue of Microscopy and Microanalysis, lower right corner of page 75, grabbing the word "Irvine" (printedpaper).
I did SE and BSE and SE+BSE of these specimens. It is interesting that the laser images turn out quite good. Not so for the offset printing. They are basically non-visible.
However, I did a series of shots using Zeiss DIC LM and can see both imprints very well. In either case, I do not see any absorption of the laser particles or the printing ink.
If anyone is interested, I can put digital images from the LM scope on the site as well. I do seem to conclude that LM observation is more useful than SEM. I would think the same about TEM.
gary g.
At 12:24 PM 8/17/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Cat-at-neto.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, August 18, 2001 at 17:24:17 ---------------------------------------------------------------------------
Email: Cat-at-neto.com Name: Jeni Hall
Organization: Alpha-Omega Home School
Education: 9-12th Grade High School
Location: Paris, Texas America
Question: If a compound microscope has a ten-power eye piece and a ninety-power objective lens, what is its magnification power? Also, what is the limitation of an optical microscope?
} Organization: Alpha-Omega Home School } } Education: 9-12th Grade High School } } Location: Paris, Texas America } } Question: If a compound microscope has a ten-power eye piece and a } ninety-power objective lens, what is its magnification power? Also, } what is the limitation of an optical microscope? } } ---------------------------------------------------------------------------
Jeni -
10x90=900. And if that is a calculation for your microscope, please be aware that any objctive of that power is going to provide a very poor image unless it is designed for use with immersion oil. You'll find a lot of well- organized,useful background information on microscope optics on the Florida State University's "Molecular Expressions" website, http://microscopy.fsu.edu/optics/index.html
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Tobias Baskin pointed out the probable difference between Gutenberg printing and modern web printing.
I was wondering about that too. Gutenberg. Heard about it but not certain what it actually is. I suspect the magazine is indeed offset web press. But if the the two processes are different, are they both based on ink of the same type? If so, I can't see it in the SEM either SE or BSE. But sure can under LM.
I suspect that based on the variety of old books I have, there may or may not be a build up of ink on the paper. Some paper is coated stock whereas others are quite porous. As I think about this, I think the reason I cannot "see" the web printing ink is because it is more "pushed" into the paper. It is akin to being absorbed by the paper. The laser toner is distinctly different and is laying on top of the paper. The laser fuser heats up the toner particles (readily visible as such) and melts and sticks the particles to the paper rather than having the paper absorb them. SEM imaging of the web printing reveals nothing but the texture of the paper fibers. The ink is embedded but is difficult to visualize using SE or BSE.
If I knew the age of Gutenberg type of printing, I do have some rather old books around. Some are late 1700's up to early 1900's. A small piece of one could be sacrificed in the name of science!
I'm working on thin layers of Au on organic substrates for doing ultrasonic imaging in catheters. I find some similarities between this and the ink question. Quite interesting and useful.
Regarding the pix at my web site-- JPEGs of same root name are at the site too. These are only about 45KB-80KB each. These may be better to access since some browsers use Quicktime to display TIFF whereas IE for example will directly display JPEG. In this case, just grab them via http://www.gaugler.com/name.jpg
For example, http://www.gaugler.com/laserpaper-2.jpg All image files are sized to be 450pixels wide.
I also added two files: printedpaper6-bsel.tif printedpaper6-bsel.jpg
which are TIFF and JPEG images of printedpaper6-bse after processing with Image Content's Lucis. I'm certainly no paper expert, but it looks like with web printing, the ink is absorbed into the paper's fibers. The lower right appears to be a glob of ink. What is puzzling is that under DIC LM, both laser and web printing have three dimensional aspects. This is not seen in SEM for both types of printing. Perhaps the Gutenberg method will indeed show up.
Moveable type was used routinely for printing from Gutenburg's invention in 1450 or so until after the second war. But even today there are many printers around who use it for small editions and other kinds of "handmade" printing projects (just as there are potters who still throw pots by hand, etc). So if anyone out there in net land is stimulated by this thread to want to image some letter press printing, you can almost certainly find a printer in your area who can give you some scraps to examine. No need whatever to cut up old books. The same action as printing from type, that is where you have metal in a frame that is inked and then a paper is pressed onto it (where the word "press" comes from) is found in other kinds of printing. I think in some methods photography plus some sort of etching is used to make the metal template. This is all contrasted to methods like the "xerox" where charge is used to put the ink to the paper (the paper in a xerox machine never gets pressed to any extent), and to methods like ink- and laser-jets where the ink is sprayed onto the paper. Again, no pressure. It is clear that the kind of ink you need to lay nicely on a raised metal surface and be pressed onto the page is different than the kind of ink you need to go through a nozzle or be transferred by charge. I can remember from hanging around my dad's letterpress office, the ink came in tins and had the consistancy of chocolate frosting. One used a putty knife to spot a glop on a glass surface and then a roller to roll it out smooth and then apply to the type. If you were printing by hand you did this after every impression--an automated press had ususally a few dozen rollers to spread the ink evenly. Anyway, I think some of these "printer's inks" are colloids of metal particles (like good old titanium oxide white house paint), so possibly imageable in the SEM, though I am not arguing against using light. (And for sure, the idea of weighing the printed output seemed excellent from the point of view of standardizing the inking quantity).
I have some students who want to examine cross sections of very porous coal char in the SEM. Trying to get bubble free blocks is proving difficult as the particles tend to float in the resin (so I am told) and it is difficult to get good impregnation. Even recoating the ground surface and repolishing leaves an unacceptable amount of bubbles in the surface and in the hollow particles. Has anyone perfected a technique for similar samples and be willing to share it? We usually have no problems with polished blocks, it is just this material!
Thanks
Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
Tina You could try making semi-permanent mounts (as long as they are stored in horizontal position) by mounting Tol blue-stained semithin sections (0.5 to about 2micrometers thick) in a drop of immersion oil. The refractive index of the oil (1.515 or thereabouts) matches that of the resin quite closely, with the result that knife marks etc disappear.
If the sections are too dry before they are mounted, the metachromicity of the Tol blue disappears, and the sections appear uniformly blue. You could try the old trick of breathing on 50deg C dried sections to slightly hydrate them before mounting in the oil. Residual liquid water in the sections is undesirable and shows up as very visible droplets and unwetted (with oil) areas.
It becomes slightly messy if you need to look at the slides with immersion optics since the coverslip then has oil below and on top. In this case you can easily remove the coverslip without problems (gently slide it off sideways), and look at the sections in oil without the coverslip.
The stain seems to last reasonably well in the oil. I don't have experimental data, but some old (few years at least) mounts still look good. The only problem is that the slides have to be stored flat.
Try it, I think you'll like it.
Regards Jan
Rinaldo Pires dos Santos wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Tina Carvalho wrote: } } } Hi, All- } } } } It sure was nice to see so many of you at M&M 2001 in Long Beach last } } week! } } } } I generally only look at 0.5 to 2.0 micrometer epoxy or acrylic sections } } in the light microscope to see where in the block I am before taking } } ultrathin sections for TEM. I stain the thick sections with toluidine } } blue, glance at them and then throw them away. However, now I want to keep } } some of them and get good photomicrographs as well. In the past when I } } coverslipped them and took (35 mm film) pictures, they looked pretty } } bad. I've heard of various fixes, such as using thicker coverslips. } } } } Now I'd like to hear from someone doing this routinely, with good } } results (if possible!). What kind of mounting media? What kind of } } coverslips? Why? What goes on with the light as it passes through the } } resin? } } } } What kind of optics? Phase? Hoffman? DIC? Plain vanilla BF? } } } } Any clues will be appreciated! } } } } Aloha, } } Tina } } Tina, } } I use with full sucess semithin sections (embedded in Spurr resin) mounted } in Canada Balsam. However, it is important to remove the resin with a } saturated solution of NaOH in ethanol, during 5 minutes (for 0.35 µm) before } the staining procedures (Toluidine Blue or another technique). The } photomicrographs taked in DIC or BF microscopies are perfect for me. } I hope to have help you. } } Dr. Rinaldo Pires dos Santos } Dept. of Botany - UFRGS } Rua Márcio Dias, 25 / apt. 305 } Bairro Nonoai } 90830-360 - Porto Alegre - RS } Brazil } E-mail: rinaldop-at-uol.com.br
-- Prof Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Hi Dave, Do you have access to a centrifuge somewhere on campus? If your samples are indeed porous and have sufficient density you could try to centrifuge the samples in your embedding media, preferably something with low viscosity. While I have never tried this, it may work. G'luk, Russ Gillmeister Microscopy Xerox Corp. RGillmeister-at-crt.xerox.com
G'day
I have some students who want to examine cross sections of very porous coal char in the SEM. Trying to get bubble free blocks is proving difficult as the particles tend to float in the resin (so I am told) and it is difficult to get good impregnation. Even recoating the ground surface and repolishing leaves an unacceptable amount of bubbles in the surface and in the hollow particles. Has anyone perfected a technique for similar samples and be willing to share it? We usually have no problems with polished blocks, it is
just this material!
Thanks
Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
Apparently Gutenberg use a proprietary ink (graphite + linseed oil + cooking? of some sort). Not surprising that it wouldn't do too well in SEM. Too much C! LM sounds good to me too. For analysis, however, it might be useful to 'dope' the ink for the study (scientific?) phase. Little colloidal gold, or silver, or osmi___um. Ought to be a song there somewhere.
Regards and happy to hear you all had such a good time,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu CASI Home Page: http://darwin.wcupa.edu/casi/ Schedule: http://darwin.wcupa.edu/cgi-bin/casireserve.cgi Please call before visiting.
} ---------- } From: Leona Cohen-Gould } Sent: Friday, August 17, 2001 3:24 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM of paper & ink } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } At 3:53 PM -0500 8/8/01, John J. Turek wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } Hello Netters, } } I have a question from a client that is beyond my area of expertise, } so I turn to you for help. } } He would like to look at the layer of ink deposited on paper by the } traditional (Guttenberg type) printing press in order to help him } standardize his printing process. I haven't a clue as to how to } approach this. His idea was to look in SEM since his understanding } is that the ink layer is, on average, 4 micrometers thick , but my } feeling is that the ink will have absorbed into the paper and not } present any topography. } } Any ideas? I have TEM & SEM at my disposal, as well as CLSM and widefield } LM. } } Thanks in advance, } Lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } }
Hi All: We are starting to look for an inexpensive, used FIB to use for materials science applications. Are there any recommendations as to where to look? Also looking for an older 200 Kv or higher TEM in good working condition. Any recommendations? Mike UT Arlington
Dear Dave, I have had some success with particles of this type. First, mix a small amount of resin and the powder thoroughly together in a small weigh boat, so that the powder is well wetted. Pour this into the bottom of your (lightly release-sprayed) mold and then carefully trickle clear resin on top of this, so your coal/resin layer stays on the bottom. You might want to try pulling a vacuum on the weigh boat sample before pouring it into the mold, as well as after all the resin is in the mold. At 12:23 PM 8/20/01 +1000, you wrote: } G'day } } I have some students who want to examine cross sections of very porous } coal char in the SEM. Trying to get bubble free blocks is proving difficult } as the particles tend to float in the resin (so I am told) and it is difficult to } get good impregnation. Even recoating the ground surface and repolishing } leaves an unacceptable amount of bubbles in the surface and in the hollow } particles. Has anyone perfected a technique for similar samples and be } willing to share it? We usually have no problems with polished blocks, it is } just this material! } } Thanks } } Dave } Regards and good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Why does numerical aperture increase with magnification, rather than decrease? In fact, why is there a relationship between magnification and numerical aperture at all? If increased numerical aperture means better light gathering ability, that implies that higher magnification objectives should produce brighter images.
This, to me, is like saying that going from a 50mm camera lens to a 1000mm lens is going to give me more light. In the photographic world, increased light gathering ability comes only from increased lens diameter and it is completely independent of the "magnification" of the lens.
I visited the Nikon N.A. Java tutorial at http://www.microscopyu.com/tutorials/java/objectives/nuaperture/index.html and this only seemed to reinforce the discrepancy for me.
With an SEM I can imagine a scan over a smaller area (a tighter scan) producing an increase in magnification. In a photographic system I can envision viewing a smaller ray bundle (smaller solid angle) producing a larger magnification. How is it that in light microscopy this is reversed and a high magnification objective takes in a larger angle? What is backward in my thinking?
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
thank you for your rapid and varied responses to my question. I think I will try the weighing trick as a quick way to get some sort of answer for this guy. Printing is his hobby, so this is not work-related, and I think he came in only because the traffic flow in the building has been redirected to pass my doors during renovation of the main entrance, and he wanted to see what an EM looks like and get a shot at playing on it. I've had to put new, coded locks on my doors because of all the curious people just wandering in! It seemed to be an interesting problem, but not one I'm willing or able to put much time/energy too. I was hoping for answers that would contain methods I'm not equipped to do (EDS/EELs, etc) and many of you sent such suggestions. Thanks again.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi, we have this problem with the Jeol 1200. It will not start, the main relay will not hold. If it starts, once in a blue moon, the forepump will or will not run. We replaced already the motion detector on the pump. If it runs, the forepump could stop after a few hours and shut down the instrument. Does anyone have any ideas, water is running well, air is good. Also I need urgently better schematics, the books are not very helpful. We are willing to pay for them. Please give us any ideas. Thank you Peter Stolzenberg, 215-699-6160 Fax 215-699-5275
Bruce, In light microscopy, there is not a quantitative relation between NA and magnification, but as you have observed the numbers are usually correlated. In theory, resolution in light micrscopy depends on the NA, not on the magnficiation. The wider angle light gathered the smaller your diffraction limited spots are comprising the image. So in theory, one could build a 1x lens with a 1.4 NA and then just enlarge the image secondarily as needed. But the trouble is that in practise, I don't believe that optical engineers could make such a lens, or at least not so that anyone could afford to buy it. To function in the low power regime it would need to image a large amount of sample and therefore the front surface of the lens would be immense (maybe as big as a 35 mm camera lens) and I don't think the glass can be manufactured that well. So as the mag goes up, the area sampled on the object goes down and this allows lenses with higher NAs to be built. Depending on the needs and the cleverness of the manufacturers, some rather nice lowish mag high NA lenses can be made. I have a 40x 1.4 NA lens that works delightfully well, and better than the 100x 1.3 NA lens by the same company.
Hope this helps a tad, TB } } } Numerical aperture/magnification/image brightness } } Why does numerical aperture increase with magnification, rather than } decrease? In fact, why is there a relationship between magnification and } numerical aperture at all? If increased numerical aperture means better } light gathering ability, that implies that higher magnification objectives } should produce brighter images. } } This, to me, is like saying that going from a 50mm camera lens to a 1000mm } lens is going to give me more light. In the photographic world, increased } light gathering ability comes only from increased lens diameter and it is } completely independent of the "magnification" of the lens. } } I visited the Nikon N.A. Java tutorial at } http://www.microscopyu.com/tutorials/java/objectives/nuaperture/index.html } and this only seemed to reinforce the discrepancy for me. } } With an SEM I can imagine a scan over a smaller area (a tighter scan) } producing an increase in magnification. In a photographic system I can } envision viewing a smaller ray bundle (smaller solid angle) producing a } larger magnification. How is it that in light microscopy this is reversed } and a high magnification objective takes in a larger angle? What is backward } in my thinking? } } Bruce Girrell } Microline Technology Corp. } 2397 Traversefield Dr. } Traverse City, MI 49686 } http://www.microlinetc.com } } (231) 935-1585 (Voice) } (231) 922-5099 (FAX) } bigirrell-at-microlinetc.com
i am doing renal bx, have been having trouble today with extreamly fine chatter. i am using araldite and an LKB nova. i can't tell if it's the building, or the microtome, or the blocks. the blocks seem the right hardness. john Hoffpauir
This message is for those of you who may not have made it to the M&M 2001 last week or missed the pharmaceutical session where we discussed FDA issues.
Jeff Hanson & Mike Esterman Scientific Imaging Center Eli Lilly and Company
************************************************************************ 21 CFR Part 11 Imaging Workshop A workshop for defining "best practices" for image data management to share with the US FDA September 24-25, 2001 Indianapolis, IN
Do you know if your image is compliant with FDA standards? Do you know the FDA's regulations regarding the appropriate handling and storage of images?
The reality is that these standards are still not defined to any degree of
certainty. That's why Eli Lilly's Scientific Imaging Center is sponsoring
a first-ever Imaging Workshop with companies in a variety of fields. This
2-day workshop will be held on September 24-25, 2001 at the University Conference Center in Indianapolis, IN. If you are involved in generating or storing scientific images that could be included in FDA submissions or in support of a submission, the 21 CFR Part 11 guideline may affect you. This workshop will attempt to document a set of best practices for handling and storage of scientific images and to suggest guidelines to the FDA for applying 21 CFR Part 11 to Scientific
Images.
Scientific Image Center Eli Lilly and Company
WHO SHOULD ATTEND This workshop is intended for scientists, QA professionals, and IT professionals who work with scientific images and imaging within industries regulated by the FDA. Vendors of instruments and software supporting their scientific imaging processes are also welcome to participate.
WORKSHOP OBJECTIVES The primary objective of this workshop will be to define a set of best practices for image data management that can be submitted as a set of recommendations to the FDA. A secondary objective will be to raise awareness of 21 CFR Part 11 and determine how it will impact scientific imaging. Finally, a post-workshop working group will be assembled to follow-up on the conclusions and continue to work through any open issues.
REGISTRATION You may send an email message to the Scientific Imaging Center at Eli Lilly and Company for additional information or to find out more about registration (ImageCtr-at-Lilly.com). Registration needs to be complete by August 31, 2001. Please note that space is limited, so we would prefer to have a single representative from each company. ************************************************************************
} Date: Mon, 20 Aug 2001 08:53:13 -0700 } To: Ike Oguocha {oguocha-at-yahoo.com} } From: Mary Mager {mager-at-interchange.ubc.ca} } Subject: Re: Choosing an Analytical SEM for Metallurgical Research } Cc: Microscopy } Dear Ike, Any of the four main manufacturers of SEMs: Hitachi, JEOL, Philips (FEI) and LEO make SEMs suitable for general metallurgical analysis. Likewise, any of the manufacturers of EDS systems, of which there are too many to name here, make systems that will suit your purposes. Make a wish list of capabilities: sample size, stage movement and automation, variable vacuum (yes or no), kV range, detectors (i.e. BSE type), EDS features and then ask the Canadian representatives of the SEM manufacturers for their information on their microscopes and a list of a few Canadian customers that you can phone or e-mail for information on the durability and service record of each of the SEMs. To write a grant application you just need a quote from each of the four, so you know how much money to ask for. Choosing exactly which SEM and which EDS can come later, after you have the grant. } At 07:03 PM 8/17/01 +0100, you wrote: } } } Hello Netters: } } } } In about three weeks time or so, I'll be writing equipment grant } } appilcations. Our lab needs, as a matter of necessity, an analytical } } SEM. The one we have now is relatively old (Philips SEM515) and } } has no EDS facilities. It hinders most of our work since we have } } to go elsewhere and pay for any work involving EDS. Our attempt } } last year to get a EDS system grant for it failed. So, we are } } making a completely new equipment grant this time. } } } } I am wondering if there are metallurgical microscopists in the } } house to give me some insights on which direction to go. What } } must we be looking for in a good analytical SEM? Who makes what? } } } } Many thanks in advance. } } } } Ike Oguocha } } Department of Mechanical Engineering } } University of Saskatchewan } } Saskatoon, Canada } } } Regards, } Mary } } Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
At 09:28 AM 8/20/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
NA does not increase with magnification. Better objectives have higher NA as they increase in magnification. The relationship between NA and magnification revolved around the issue of resolution (resolving power).
The greater the resolving power is, the smaller the distance between objects that can be distinguished. Resolving power is:
RP=Lambda/2NA
where Lambda is the wavelength of light. So, to increase RP, decrease wavelength and/or increase NA. Decreased wavelength is the same as increasing frequency. White light is a combination of all colors (many different wavelengths). Removing most the wavelengths via a filter (IR, for example) will improve RP. The "frequency" of an electron beam is super high and thus, has extraordinary RP.
Brightness is mostly due to the amount of glass in a lens. For high brightness and high NA, use a PlanAPO objective. Bring lots of money.
} This, to me, is like saying that going from a 50mm camera lens to a 1000mm } lens is going to give me more light. In the photographic world, increased } light gathering ability comes only from increased lens diameter and it is } completely independent of the "magnification" of the lens.
A 50mm/f2 lens will have the same brightness as a 1000mm/f2 lens. Unfortunately, f-stop is the ratio of the focal length divided by the diameter of the focusing element. So, a 50mm/f2 lens would have to have a front lens element which is about 25mm in diameter. The 1000mm/f2 lens would have a front element which is 500mm in diameter. This huge piece of glass would weigh about five pounds and the whole lens would cost about $15,000. My Nikkor 600mm/2.8 AI-EDIF weight about thirty pounds and cost $8500. It is definitely not for point and shoot events.
The alternative to refractive lenses are reflective or Catadioptric "lenses." These are mirrors, like large telescopes. They have fixed f-stop figures and are small and light. But the typical f-stop limit is f8.
} I visited the Nikon N.A. Java tutorial at } http://www.microscopyu.com/tutorials/java/objectives/nuaperture/index.html } and this only seemed to reinforce the discrepancy for me. } } With an SEM I can imagine a scan over a smaller area (a tighter scan) } producing an increase in magnification. In a photographic system I can } envision viewing a smaller ray bundle (smaller solid angle) producing a } larger magnification. How is it that in light microscopy this is reversed } and a high magnification objective takes in a larger angle? What is backward } in my thinking?
A SEM offers huge depth of field (high working distance) at low to medium magnification. This is due to many factors. One is the high frequency of the electrons and the extreme collimation of the electron beam (not a lot of extraneous energy hitting the specimen). SEMs still suffer from spherical abberations and astigmatism. This is diminished by stigmator coils. For optical lenses, it is solved pretty much by lots of money at time of purchase.
I agree with Gary Gaugler's description (the entire reply is appended below) but let me add one minor refinement to his comment : " Brightness is mostly due to the amount of glass in a lens. For high brightness and high NA, use a PlanAPO objective. Bring lots of money."
I agree with this for brightfield but for high brightness with UV (e.g., calcium sensitive fluroochromes like Fura or DAPI), the large amount of glass in a highly corrected PlanApo works against brightness. A high NA achromat can sometimes be a better choice if you don't have a big field of view.
} } -----------------------------------------------------------------------. } } } Let me try to give it a stab-- } } At 09:28 AM 8/20/2001, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
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Try the following trick: turn OFF microscope as usual, wait until all lights is OFF on the panel, than disconnect instrument from main power source (we do have some huge breaker on the wall), turn ON microscope as usual.
Good luck, Sergey
At 02:56 PM 8/20/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Any short run publication pre 1950 can be assumed to be printed on letter press. Duplicate journals would be a good place to look. Anything printed by the college pre 1950 will fall in that category as well unless it was running a experimental print shop. A stop by the university print shop should get you some modern samples of printing and ink. They still probably do some things that way or know some one that does. It still is hard to beat for short runs on weird shaped stuff.
You can look at some old books under a LM and see what letter press looks like and sort though the modern stuff. Letter press hasn't changed enough in the last 100 years to show up much different under a LM.
To get much of a idea of what the film actually looks like I think you will have to section the page edgeways.
One advantage of using pre 1950 inks is that there will probably be lead in them and that should increase the contrast. I think many of the pigments and dryers are metals so you might get x-rays fluorescing out of them as well at high voltage.
Since 1950 more and more attention has been paid to safety in the printing industry. That translates in to less use of heavy metals in inks. So old inks are likely to image better without enhancement than modern inks. Opaque blue and green inks would probably be good choices. Cobalt was used in blue ink until as late as 1970 and I haven't kept up since then.
If you can't find samples I can find some send them to you.
The inks are not a lot different between the two processes. Letter press ink will work in an offset press and visa versa but they will not work as well as they will in the press they are designed for. Black printing ink can be made by grinding carbon black in boiled linseed oil. Form there the chemistry gets increasingly complicated. Today inks are usualy soy bean oil based due to health concerns.
Coated papers will have more ink build up than uncoated papers. Book papers that don't appear to be coated have considerable sizing to keep the ink from being absorbed in the paper fibers and being seen from the back side. This paper is also loaded with clay to increase it's opacity.
Gordon ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Cc: "Tobias Baskin" {BaskinT-at-missouri.edu} Sent: Sunday, August 19, 2001 1:49 PM
OK.....sounds like I am back to cutting up an old book.
I have found some modern samples of letter press from Tobias' references. But as you point out, the modern inks may not be the same as yesteryears'.
I shot a check account number today and was rather under whelmed. It is not on the surface like the laser print. I think that the challenge of measuring ink on paper is not all that easy. Probably not a big surprise to most folks. But the study of thin films is very useful and translatable to many other products.
Interesting and useful topic. Perhaps, not seemingly so.
gary g.
At 07:26 PM 8/20/2001, you wrote:
} Any short run publication pre 1950 can be assumed to be printed on letter } press. Duplicate journals would be a good place to look. Anything } printed by the college pre 1950 will fall in that category as well unless it } was running a experimental print shop. A stop by the university print shop } should get you some modern samples of printing and ink. They still probably } do some things that way or know some one that does. It still is hard to beat } for short runs on weird shaped stuff. } } You can look at some old books under a LM and see what letter press looks } like and sort though the modern stuff. Letter press hasn't changed enough } in the last 100 years to show up much different under a LM. } } To get much of a idea of what the film actually looks like I think you will } have to section the page edgeways. } } One advantage of using pre 1950 inks is that there will probably be lead in } them and that should increase the contrast. I think many of the pigments and } dryers are metals so you might get x-rays fluorescing out of them as well } at high voltage. } } Since 1950 more and more attention has been paid to safety in the printing } industry. That translates in to less use of heavy metals in inks. So old } inks are likely to image better without enhancement than modern inks. Opaque } blue and green inks would probably be good choices. Cobalt was used in blue } ink until as late as 1970 and I haven't kept up since then. } } If you can't find samples I can find some send them to you. } } The inks are not a lot different between the two processes. Letter press ink } will work in an offset press and visa versa but they will not work as well } as they will in the press they are designed for. Black printing ink can be } made by grinding carbon black in boiled linseed oil. Form there the } chemistry gets increasingly complicated. Today inks are usualy soy bean oil } based due to health concerns. } } Coated papers will have more ink build up than uncoated papers. Book papers } that don't appear to be coated have considerable sizing to keep the ink from } being absorbed in the paper fibers and being seen from the back side. This } paper is also loaded with clay to increase it's opacity. } } Gordon } ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Cc: "Tobias Baskin" {BaskinT-at-missouri.edu} } Sent: Sunday, August 19, 2001 1:49 PM } Subject: Re: TEM of paper & ink } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Tobias Baskin pointed out the probable difference between } } Gutenberg printing and modern web printing. } } } } I was wondering about that too. Gutenberg. Heard about it } } but not certain what it actually is. I suspect the magazine is } } indeed offset web press. But if the the two processes } } are different, are they both based on ink of the same type? } } If so, I can't see it in the SEM either SE or BSE. But sure } } can under LM. } } } } I suspect that based on the variety of old books I have, } } there may or may not be a build up of ink on the paper. } } Some paper is coated stock whereas others are quite } } porous. As I think about this, I think the reason I cannot } } "see" the web printing ink is because it is more "pushed" } } into the paper. It is akin to being absorbed by the paper. } } The laser toner is distinctly different and is laying on top } } of the paper. The laser fuser heats up the toner particles } } (readily visible as such) and melts and sticks the particles } } to the paper rather than having the paper absorb them. } } SEM imaging of the web printing reveals nothing but the } } texture of the paper fibers. The ink is embedded but } } is difficult to visualize using SE or BSE. } } } } If I knew the age of Gutenberg type of printing, I do have } } some rather old books around. Some are late 1700's up } } to early 1900's. A small piece of one could } } be sacrificed in the name of science! } } } } I'm working on thin layers of Au on organic substrates } } for doing ultrasonic imaging in catheters. I find some } } similarities between this and the ink question. Quite } } interesting and useful. } } } } } } Regarding the pix at my web site-- } } JPEGs of same root name are at the site too. These are } } only about 45KB-80KB each. These may be better to access since } } some browsers use Quicktime to display TIFF whereas } } IE for example will directly display JPEG. In this case, } } just grab them via http://www.gaugler.com/name.jpg } } } } For example, http://www.gaugler.com/laserpaper-2.jpg } } All image files are sized to be 450pixels wide. } } } } I also added two files: } } printedpaper6-bsel.tif } } printedpaper6-bsel.jpg } } } } which are TIFF and JPEG images of printedpaper6-bse } } after processing with Image Content's Lucis. I'm certainly } } no paper expert, but it looks like with web printing, the } } ink is absorbed into the paper's fibers. The lower right } } appears to be a glob of ink. What is puzzling is that } } under DIC LM, both laser and web printing have three } } dimensional aspects. This is not seen in SEM for } } both types of printing. Perhaps the Gutenberg method } } will indeed show up. } } } } gary g. } } } } } }
If you are really interested in comparing methods having a shop print up samples with both presses using the same color that has a metal oxide as a pigment would probably be the best approach. A small shop could do that in 30 minutes time by washing up the two presses and reinking them and then cleaning them up again.
Get some samples of the various colors of ink they have an see how they image first. One or two should have pretty good contrast. If they don't you can make your own ink by mixing what every powder you want with transparent base and ink with no pigment. Metal powders don't work very well. They don't stay dispersed it might be OK for imaging but it sure is hard to get them to look good to the eye.
If you mix in your own pigment make sure they wash the press as soon as they are done. Most inks they use today dry very slowly on the press and some pigments are natural dryers and can cause the ink to dry on the press in an hour or less. Reds are particularly bad actors in this regard.
Gordon
----- Original Message ----- } From: "Monson, Frederick C." {fmonson-at-wcupa.edu} To: "'Microscopy Listserver'" {microscopy-at-sparc5.microscopy.com} Cc: "'Leona Cohen-Gould'" {lcgould-at-med.cornell.edu} Sent: Monday, August 20, 2001 9:55 AM
If the particles are "small" then the previous suggestion to centrifuge ought to work well. If the particles, however, have lots of pores, then vacuum infiltration would be my choice.
Nothing works if the pores are truly sealed, as in pumice, but most difficult specimens can be infiltrated with vacuum. There is an easy method and a good method, but easy may be good enough: Easy: 1 Place specimen in suitable container and the container into a larger container to take any "mess". 2 Cover specimen with a low viscosity, undiluted resin; if required weigh down specimen. Place into a vacuum chamber. Evacuate briefly, stop evacuation just before resin's boiling point. Vent chamber and then re-evacuate.
Note boiling creates bubbles. The method pulls air out of the specimen and the resin is forced in when venting.
In the "good" method the specimen is evacuated first and the resin is added while under vacuum. This avoids most bubbles in the resin and takes more air out of the pores. Apparatus for this is available, but for small scale work I used a very simply modified vacuum desiccator.
"Good method": 1 As in one above, weigh specimen down. 2 Find a rubber bung that would seal the top valve hole in a desiccator lid (or cut a suitable hole in a small plastic desiccator lid). Cut a hole into the bung that would take the stem of a plastic Pasteur pipette. 3 Find a very large capacity (bellow type) polyethylene Pasteur pipette. Fill pipette with "sufficient" resin. 4 Insert stem of pipette through the hole in that bung, have only a couple of cm protrude past the smaller end of the bung. Kink and apply a bulldog clip to the pipette stem above the upper end of the bung. 5 Evacuate the specimen using the side port of the desiccator. Close the vacuum valve and remove the bulldog clip. The resin should be pulled into the chamber and run over the specimen. 6 Leave under vacuum for a couple of minutes and then vent.
Hope this helps. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Monday, August 20, 2001 12:24 PM, Dave Felon [SMTP:emudp-at-mail.newcastle.edu.au] wrote: } } G'day } } I have some students who want to examine cross sections of very porous } coal char in the SEM. Trying to get bubble free blocks is proving difficult } as the particles tend to float in the resin (so I am told) and it is difficult } to } get good impregnation. Even recoating the ground surface and repolishing } leaves an unacceptable amount of bubbles in the surface and in the hollow } particles. Has anyone perfected a technique for similar samples and be } willing to share it? We usually have no problems with polished blocks, it is } just this material! } } Thanks } } Dave } } } } } Dave Felon } EM/X-Ray Unit } University of Newcastle } NSW 2308 } AUSTRALIA } Ph 02 4921 5667 } Fax 02 4921 7019 } emudp-at-mail.newcastle.edu.au
I need some advice on a new vacuum coater for a TEM lab. I presently have a 13 year old Denton 502A system which has performed satisfactorily. Our main use is for thermal evaporation of Au, C and Al. Denton no longer sells this model but has a new system, an "Explorer 14". I have also gotten a quote for an Edwards "Auto 306" which I know little about. Are there other systems I should be considering? I was considering diffusion systems for reliability. Are turbo systems reliable, and agressive enough now? I would appreciate any comments on the performance of these or other systems. Please respond directly. Thanks for your time. Russ Gillmeister Microscopy Xerox Corp. RGillmeister-at-crt.xerox.com
Hi Bruce, I've done what I think is correct below in red - just in case it's wrong.
} ---------- } From: Bruce Girrell } Reply To: bigirrell-at-microlinetc.com } Sent: Monday, August 20, 2001 12:28 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM - Something I just don't get } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Numerical aperture/magnification/image brightness } } Why does numerical aperture increase with magnification, rather than } decrease? In fact, why is there a relationship between magnification and } numerical aperture at all? } } } I don't remember an equation relating N.A. and magnification. The only one I ever knew was Resolution=(0.61*Wavelength)/index of refraction * sine alpha.{Don't you just love text} Numerical aperture=index of refraction * sine alpha! If you consider the meaning of this, you will readily see that N.A. is dependent on the one had on the optical character of a material between object and objective lens and alpha is the half angle of the aperture angle of the objective lens. The latter can be modified by the designing engineer. All of the above derives from a consideration of diffraction by the specimen and capacity of a lens as defined by its geometry.
} If increased numerical aperture means better } light gathering ability, that implies that higher magnification objectives } should produce brighter images. } } } But what is implied if you CAN get a well-illuminated and highly magnified image from a dark object such as a section of plant stem? A bright back light, of course. Everything in the optical axis is designed to take advantage of the design characteristics of the imaging system. The light is not only bright but focused by the condenser. The specimen is transparent. Oil is added to increase the index of refraction between objective and object (then, as a afterthought, between object and condenser).
} This, to me, is like saying that going from a 50mm camera lens to a 1000mm } lens is going to give me more light. In the photographic world, increased } light gathering ability comes only from increased lens diameter } } } What is your purpose when you change photographic lenses, and why does the higher power lens have to be of greater diameter? This is the key, I think. Remember, the first compound microscope? The first guy who bought one put the objective to his eye and cursed. The compound microscope is engineered [and so is the camera lens]. Why? Different purpose! Tube length, lens geometry, working distance, depth of focus, etc. are engineered to perform a task. Further, in the microscope, magnification is related to working distance and lens geometry. Light gathering capacity is then determined by the illuminator and the condenser (i.e., how much of the available light can be focused on the "field of view". The confocal microscope was patented in the '60's. It was a brilliant idea without a proper light source. } and it is } completely independent of the "magnification" of the lens. } } } I think from what I said above, it is clear that N.A. is NOT dependent on mag., but it IS often described as the capacity of a lens to gather light. Alpha, in the equation above is really limited by the geometry of the objective lens, but the intensity of the illumination is dependent on the geometry (and adjustment) of the condenser as well as the power of the "light". All of this is necessary, because the object is NOT a(the) light source [we have to use Kohler logic to fool it], while in the use of the camera, we depend on the back or "ambient" light or an added "reflected light" for illumination.
} I visited the Nikon N.A. Java tutorial at } http://www.microscopyu.com/tutorials/java/objectives/nuaperture/index.html } and this only seemed to reinforce the discrepancy for me. } } With an SEM I can imagine a scan over a smaller area (a tighter scan) } } } Why don't we do it that way then? } producing an increase in magnification. In a photographic system I can } envision viewing a smaller ray bundle (smaller solid angle) producing a } larger magnification. } } } Haven't you just described the function of the front lens without considering the functions of the others behind it? } How is it that in light microscopy this is reversed } and a high magnification objective takes in a larger angle? } } } Again, I think it is all about the geometry of the objective lens and the intent of the optical engineer. } What is backward } in my thinking? } } } Nothing! Did you ever get a question on an exam that had four story lines only one of which is relevant? Here, you are trying to understand a microscope by studying a camera. They are DESIGNED to achieve different outcomes. Practical question. Have you ever heard of anyone using a camera lens to achieve higher magnification with a microscope or a microscope objective to get higher "zoom" with a camera? Neither have I.
} } } [When I was just a freshman in college, I was advised to change my major from Biology to Mathematics. I was a biology major, I explained, because I didn't have sufficient confidence in my mathematical acumen to be a Chemistry major (one "A" in PChem every 4-5 years) which ought to explain why I wouldn't consider switching to Math. Thank God for the Slayter's (see below) of this world who have taken on the task of instructing me in spite of my computational weaknesses.]
There is a GREAT book by one Elizabeth Slayter entitled "Optical Methods in Biology", Wiley-Interscience, NY, 1970. There is a more recent edition. I have used her instruction to provide help on questions whose answers I should remember from physics.
Regards and sorrow for forcing you to suffer through my thinking on the fly,
Fred Monson
} Bruce Girrell } Microline Technology Corp. } 2397 Traversefield Dr. } Traverse City, MI 49686 } http://www.microlinetc.com } } (231) 935-1585 (Voice) } (231) 922-5099 (FAX) } bigirrell-at-microlinetc.com } } }
Just to summarize some of the information regarding NA and Resolution and Magnification: Resolution and numerical aperture are related by resolution = wavelength/N.A. objective + N.A. condenser. Brightness, numerical aperture and magnification are related by brightness= (N.A.)squared /(magnification)squared. As is apparent from these equations, in an idealized situation, a larger numerical aperture at a given magnification will yield better resolution and greater brightness. Things get a little more complicated in real life as has been astutely pointed out, however. Glass has different refractive indexes for different wavelengths of light. White light is composed of red, green and blue wavelengths, and this presents a problem when specimens are illuminated with full spectrum light sources. The red, green and blue components of the image will be refracted by the lens at slightly angles, and so the focal points for the different wavelengths end up being slightly different and the result is that objects appear to be surrounded by a color fringe. This phenomenon, called axial chromatic aberration, is exasperated at greater angles incident to the lens. In other words, at high mag, the higher the numerical aperture the more pronounced the chromatic aberration. It's a concept that is easy to illustrate but hard to explain. The solution, put simply, is to combine different types of glass which have equal but opposite differences in refractive index for light at different wavelengths. This involves more glass, more smart people with handsome salaries to engineer the glass, and results in an expensive plan-apochromatic lens which is corrected for chromatic aberration in the red, green and blue spectra. If the application only requires limited spectra, as in ultraviolet illumination, chromatic aberration isn't such a concern, and one can get by with less expensive lenses of the same numerical aperture, and because of less corrective optics, may indeed be transmit more light to the eye. Spherical aberration is a defect of lenses in which the surface forms part of a sphere. High numerical apertures at high magnification exasperate spherical aberration, so high quality lenses incorporate corrective optics to bring areas on the circumference of the field of view into focal register with the center of the field of view. Again, more glass, more smart people, more money. So the long and the short of it are that high mag lenses don't necessarily have high numerical apertures, however resolving power and brightness will suffer proportionally. High numerical aperture high mag lenses may incorporate artifact into the image unless they are engineered with compensating corrections. Some of these artifacts, which are expensive to correct, may not be an issue under certain circumstances, and may even hinder optical performance, so it is important to understand what corrections one is purchasing, and why.
Karl G.
/**************** Karl Garsha www.uwm.edu/~keg ****************/ ----- Original Message ----- } From: "Bruce Girrell" {bigirrell-at-microlinetc.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, August 20, 2001 11:28 AM
Hi all
How stable are LRWhite sections? Can I cut slices of LRWhite today and give them to a student lab to do immunogold labelling next week, without the slices losing or altering their antigen-antibody affinity or sensitivity?
I have found epon sections stain better with uranyl acetate immediately after being cut, as compared to uranyl acetate staining several days later (Lead staining doesn't seem to be similarly affected). This suggests there are alterations taking place in the plastic slices between slicing and some time period afterwards. Does anyone have any similar anecdotes for LRWhite, specifically in regard to antibody labels? That is, does labelling efficiency drop off over time in plastic embedded sections?
look forward to hearing from someone in this regard
Steve --
___________________________________________________ Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
Here's one I've never encountered. A colleague just brought me several roundish, dark brown to black particles, about a millimeter in diameter, taken from his car. Although they seem fairly hard, they can be cut with knife, leaving a waxy smear on the microscope slide. Some have flat sides from where they adhered to the car surface. All have a wrinkled, somewhat bubbled looking texture. The exterior appears to be slightly soluble in toluene. When subjected to a hot wire, a brown, oily substance cooks off, leaving a charcoal-like core. The question is, could this be an emission product coming from a large diesel generator that was just put into operation at our site?
The Particle Atlas doesn't depict anything quite this large, so I had some reservations. Much obliged for any thoughts.
In our hands and for our application the Abcam rabbit polyclonal to green fluorescent protein worked pretty good in immuno-em with pFA fixed samples. You will find informations about this antibody at: http://www.abcam.com
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer MCDB Dept. Yale University P.O. Box 208103 New Haven, CT 06520-8103 U.S.A.
Hi Ike, Mary got the SEM info right on since she mirrored our experience with writing a proposal. With respect to EDS I have only one recommendation. We all depend on the big four: Oxford, EDAX, PGT and Thermo-Noran. We are also considering two or three others: IXRF, 4pi Analysis, Quartz X-Ray, Rontek and Clemex. These run the gamut from PC-based packages to off-the-shelf components. We are looking at these, because they are considerably less expensive than the big four and, because we are trying to understand exactly what it is we want to do with EDS. Also, do we want WDS in addition or instead? There is a plethora of information on the subject of EDS on the net, but there is nothing that will replace a thoughtful consideration of three lists: needs, wants and wishes. At every level of a grant proposal, a clear understanding of how to justify each type of component. Though perhaps not considered a mechanical engineering area, you might do a quick scan of a methodology called EBSD (electron backscatter diffraction).
With respect to the SEM, there is the expensive path and the CAMSCAN (http://www.camscan-usa.com/VEGA%20specification.htm#Title) VEGA Series of variable pressure systems. Include the array of SEM substages available from Fullam (http://www.fullam.com/Sem_subs.htm#18025), and you have almost every piece of info you need. The only extra piece of information you may have to consider is how much variability in pressure you might NEED to accomplish the goals set aside for the new system. Good luck and hope you get it.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu CASI Home Page: http://darwin.wcupa.edu/casi/ Schedule: Navigate from CASI Home Page Please call before visiting.
} ---------- } From: Ike Oguocha } Sent: Friday, August 17, 2001 2:03 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Choosing an Analytical SEM for Metallurgical Research } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Netters: } } In about three weeks time or so, I'll be writing equipment grant } appilcations. Our lab needs, as a matter of necessity, an analytical } SEM. The one we have now is relatively old (Philips SEM515) and } has no EDS facilities. It hinders most of our work since we have } to go elsewhere and pay for any work involving EDS. Our attempt } last year to get a EDS system grant for it failed. So, we are } making a completely new equipment grant this time. } } I am wondering if there are metallurgical microscopists in the } house to give me some insights on which direction to go. What } must we be looking for in a good analytical SEM? Who makes what? } } Many thanks in advance. } } Ike Oguocha } Department of Mechanical Engineering } University of Saskatchewan } Saskatoon, Canada } } } ____________________________________________________________ } Do You Yahoo!? } Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk } or your free -at-yahoo.ie address at http://mail.yahoo.ie } }
I have a question regarding breaking glass knives with the LKB 7800 knife-breaker. When I was trained on the instrument, my advisor threatened me with instant death if I ever tried to break 8mm thick glass strips rather than the standard 6.4mm we used. Now I have one of my own and I have a faculty member who would like to use the thicker glass. Has anyone used 8mm glass with the 7800 successfully, or was my advisor correct?
Thanks for your help
Todd
-- Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility Department of Biology and Microbiology University of Wisconsin Oshkosh 800 Algoma Blvd Oshkosh, Wisconsin 54901 Ph: 920-424-3069 Fax: 920-424-1101 E-mail: kostman-at-uwosh.edu
I'd say it probably depends on the antigen - I've had blocks that "went bad" for a particular antibody within a month or two, but I've also used grids of LRWhite sections that had been cut weeks/1-2 months in advance. (This was for a course, so we were already stacking the deck in our favor by using antibodies that labelled really strongly so the students would see something when they did their labelling - the sections were cut in advance for them). The intensoity was somewhat reduced, but that may have been a function of the course environment...things that normally work well tend to weird out.
Histology people say the same thing; some samples can be cut in advance and the sections stored, others are more susceptible to air exposure. If you can keep the sections cold they may be "happier". Cryosections (for EM) can apparently be stored for months at 4C in methylcellulose and be just fine!
Tamara
On Tue, 21 Aug 2001, Steve Barlow wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all } } How stable are LRWhite sections? Can I cut slices of LRWhite today } and give them to a student lab to do immunogold labelling next week, } without the slices losing or altering their antigen-antibody affinity } or sensitivity? } } I have found epon sections stain better with uranyl acetate } immediately after being cut, as compared to uranyl acetate staining } several days later (Lead staining doesn't seem to be similarly } affected). This suggests there are alterations taking place in the } plastic slices between slicing and some time period afterwards. Does } anyone have any similar anecdotes for LRWhite, specifically in regard } to antibody labels? That is, does labelling efficiency drop off over } time in plastic embedded sections? } } look forward to hearing from someone in this regard } } Steve } -- } } ___________________________________________________ } Dr. Steven Barlow } EM Facility/Biology Dept. } San Diego State University } 5500 Campanile Drive } San Diego CA 92182-4614 } phone: (619) 594-4523 } fax: (619) 594-5676 } } email: sbarlow-at-sunstroke.sdsu.edu } http://www.sci.sdsu.edu/emfacility } } Chairman, Educational Outreach subcommittee } promoting microscopy instruction and increased access to microscopes } Microscopy Society of America } http://www.msa.microscopy.com/ } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
I'm using the Image Processing Toolkit for Adobe Photoshop to calculate areas and perimeters of SEM images and was wondering if anybody could provide a simplified explaination of convex area and convex perimeter versus just area and perimeter.
Please, respond offline.
Sincerely,
Cavin Mooers, Research Associate Vitreous State Laboratory The Catholic University of America Hannan Hall Washington, DC 20064 (202) 319-5346phone (202) 319-4469fax
The Nuclear Materials Science Group (NMT-16) of the Nuclear Materials Technology (NMT) Division at Los Alamos National Laboratory is seeking an experienced Surface Scientist interested in the complex surface chemistry of plutonium and other actinide metals. The successful candidate will lead the surface science effort in the Microstructure and Microanalysis team. Working independently as the subject matter expert, this individual will plan and execute experiments in support of the Pit Manufacturing, Pit Surveillance and Certification Programs. They will have for their use two newly acquired instruments: a JEOL JAMP-7830 field-emission gun Auger Microprobe (Auger, XPS, cold fracture, heating, orientation imaging microscopy, gas reaction, AFM/STM); and a Kratos Axis-Ultra imaging XPS system (XPS, Auger, heating/cooling, gas reaction). These instruments will be installed in the Plutonium Facility at TA-55 in the summer of 2002, and enhance the experimental capabilities of a well-equipped material science laboratory that includes: variable pressure SEM (EDS, WDS, OIM, CL spectroscopy, hot/cold stage); new 5 spectrometer JEOL 8200 Electron Microprobe; diverse x-ray diffraction capabilities; hardness testers; several modern optical microscopes with digital cameras; and a nicely equipped metallography line.
Required Skills: Prior hands-on experience gathering and interpreting Auger and XPS data. Experience maintaining state-of-the-art UHV materials characterization equipment. Practical and theoretical knowledge of Auger and photoelectron spectroscopy. Recent peer-reviewed publications that include Auger and XPS as characterization tools. Working knowledge of quantification procedures relating to these techniques. Strong organizational skills as evidenced by the flexibility to work on multiple tasks simultaneously. Excellent communication skills. Ability to obtain a Q clearance, which normally requires U.S. citizenship.
Desired Skills: Experience in preparing radioactive materials for analysis. Demonstrated ability to handle and work with hazardous chemicals in a safe manner. Considerable knowledge and experience in any of the following technical areas: handling and conducting experiments with actinide metals, alloys and compounds; working in labs containing radioactive materials; and applying quality assurance requirements to sampling and analysis activities. Experience in gathering and interpreting technical information and reporting technical results both orally and in writing. Active Q clearance and PSAP.
Education Requirement: Ph.D. degree in Materials Science, Physics, Chemistry or equivalent combination of relevant education and experience.
Additional Requirements: This position is subject to the requirements of the Personnel Security Assurance Program (PSAP). Candidates invited to interview for this position will be subject to a pre-employment screening check, medical examination and drug test, and must consent to be in the program at the time of the interview.
The position described above will be posted at the official LANL Human Resources Department's web page for job openings. Any individual interested in this position MUST apply through this web site to be formally recognized as an applicant. The HR web site also has guidelines for TSM salaries. Any questions about this opening can be directed to: Brad Storey (505) 667-0458 storey-at-lanl.gov or Rollin Lakis (505) 665-9814 rlakis-at-lanl.gov
*************************************************** Brad Storey, Ph.D. Team Leader, Microstructure & Microanalysis NMT-16 - Nuclear Materials Science Group Nuclear Materials Technology Division Los Alamos National Lab PO Box 1663, MS E574 Los Alamos, NM 87545
I am at the point of deciding whether to dump my Axioplan system for an Olympus BX-51. I'm not sure that this is a good idea or is a decision in a positive direction. Thus, I seek your input on this issue.
As I mentioned before, my current Zeiss system is not working beyond about 200X. Is there someone in the Sacramento CA region who can work on my system and find out what is wrong with it? If some minor tweak is all that is necessary, I certainly don't need to spend another $40K for a new 'scope. If I can take the system to someone nearby, that would be good too.
I have the basic following items:
Axioplan 1 stand 44 72 15 100W lamphouse 2ea 44 53 65 condenser turrets 44 52 48 Ph/DIC turret condenser w/ 46 52 68 1.4 top lens 44 53 51 auto-swingout lens assembly with 0.9NA top lens 44 53 50 manual swingout lens assembly with 0.9NA & 1.4 top lenses
Plan Neofluar 1.25/0.075 Plan Neofluar 20X/0.5 Ph2 Acrostigmat 100X/1.25 Ph3 Plan Neofluar 40X/0.75 Ph2 Plan Neofluar 63X/1.25 oil Plan Neofluar 20X/0.5 with DIC prism Plan Neofluar 40X/0.75 with DIC prism Plan Neofluar 40X/1.30 oil Plan Neofluar 5X/0.15 Plan Neofluar 10X/0.30 Plan Neofluar 63X/1.25 oil with DIC prism Plan Neofluar 100X/1.30 oil with DIC prism 44 4 80
Is there any chance for support and/or parts for this system? Is it a lost cause? You indicated that these systems were still supported. How can I get such support?
gary gaugler
At 03:58 PM 7/25/2001, you wrote:
} Dear Dr. Gaugler,
} I want to thank you for taking the time to talk to me yesterday. I appreciate } your honesty and frankness. We are always glad to hear from our customers, } especially if the feedback is constructive. Certainly, I was pleased to hear } your praise for the performance our Microscopes deliver. Of course, I was } surprised to hear of your negative experience with our support } team. Therefore, } I would like to answer your email and phone conversation in two parts:
We would like to order some conductive carbon cement (Leit-c), but were told that we have to pay more than $100 for special package over the price of the cement because of the chemical contents in the cement. We wonder who has bought Leit-C recently and through which local dealer in Australia. Thanks for any information. Cheng Huang
----------------------------------------------------- Cheng Huang Australian National University EM Unit, RSBS Box 475, ACT 2601 Canberra, Australia Phone: 61-2- 6125-6553 Fax: 61-2- 6125-3218 http://www.anu.edu.au/EMU/
} From what I have seen coming out of diesel engines it is soot from unburned fuel. This is not a scientific observation but the observation of a farming. It comes from running the fuel too rich and in high load conditions. Taken to extremes I have seen a tractor blowing a plume of black smoke from 12 miles away on a still morning.
Regulations no longer allow setting like this but you can still see engines putting out black smoke when starting up from a stop. It is more pronounced in older trucks. The soot particles can be fairly large.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger ----- Original Message ----- } From: {"David_R_Stadden-at-armstrong.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 21, 2001 1:43 PM
In a message dated 8/21/01 9:58:12 PM, cavinm-at-vsl.cua.edu-at-sparc5.microscopy.com writes:
} I'm using the Image Processing Toolkit for Adobe Photoshop to } calculate areas and perimeters of SEM images and was wondering } if anybody could provide a simplified explaination of convex area } and convex perimeter versus just area and perimeter.
Thought that was covered in the tutorial on the CD. Anyway, the convex area and perimeter are measured by constructing a bounding polygon (32 sided in the tool kit) around the feature. This is sometimes called a "taut string" or "rubber band" outline since it does not follow any indentations in the periphery of the object. For a convex shape with no internal holes, the values are identical to the "regular" perimeter and area.
I can offer a story I heard at a meeting. This is actually about a factory where cameras were assembled. Contamination on the lenses was subjected to EDX and the elements seen (Na?, K?) were consistent with saliva. Rather than ban talking, the management solved the problem by use of transparent screens above the equipment.
Dave
On Wed, 15 Aug 2001 07:56:07 -0400 Peter Tomic {PTomic-at-anadigics.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone out there know of a good source, text or website, that deals } with the issue of contaminants in a semiconductor manufacturing environment? } I'm putting together a lecture on the subject but I don't want to generate } SEM's, optical images etc. if this is available readily. Principally I'm } interested in human spittle, airborne contaminates such as skin, hair, cloth } fibers, etc. or anything that may be generated by human interaction with a } semiconductor device. } } Regards, } Peter Tomic } Anadigics, Inc. }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
1) tighten the block in the chuck 2) tighten the knife in its holder 3) check knife angle(I use 4 deg. clearance angle) 4) try at different times of the day- at my old place my lab was next to two large turbines and early in the day I occasionally had problems with chatter
Hi all, Does anyone know the proper bulb for a Durst 138S Laborator enlarger? Is it a giant 110V, 200W opale Atlas bulb? and if so where can I get one? Durst has not been very helpful. thanks for the help, Beth
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
Hi Todd, I have a 7800B, and an official "LKB 7800 KnifeMaker Spare Parts Catalog". In all of the spare parts and in all of instructions which we also still have, there is NO mention of glass with thickness other than 6-7mm. I have always taken that to mean, "NO THICKER GLASS ON PENALTY OF DEATH!" Since we kept a 1/2" Dupont knife breaker, there was no need to pester folks about this, I merely kept the 1/2" glass close to the Dupont, the "DEATH" notice over the LKB and depended on human nature for the rest. I was correct, no one was ever seen to carry 1/2" glass the 3 feet from the Dupont to the LKB. I was always easier to reach up and get a strip of the 6mm from the supply. Also, if 1/2" glass gave better knives than 1/4", none of us would have kept the LKB's - I think! I have tried the 1/2" from the Dupont on an ultramicrotome, and the results were spotty. I thought that the Dupont broke better thin-sectioning knives from 1/4"(6mm) glass. Get the man(?) an estimate, and don't use the LKB for something for which it apparently not designed. Watch me get followed by someone who breaks 1/2" glass all the time with no visible harm to the LKB or the 6mm knives. Well, I'm old enough to refuse to change and just be called a "Grumpy Old Man".
Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu CASI Home Page: http://darwin.wcupa.edu/casi/ Schedule: Navigate from CASI Home Page Please call before visiting.
} ---------- } From: Todd Kostman } Sent: Tuesday, August 21, 2001 4:37 PM } To: Microscopy Listserver } Subject: Knifebreaking } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } } I have a question regarding breaking glass knives with the LKB 7800 } knife-breaker. When I was trained on the instrument, my advisor } threatened } me with instant death if I ever tried to break 8mm thick glass strips } rather } than the standard 6.4mm we used. Now I have one of my own and I have a } faculty member who would like to use the thicker glass. Has anyone used } 8mm } glass with the 7800 successfully, or was my advisor correct? } } Thanks for your help } } } Todd } } } -- } Dr. Todd A. Kostman } Assistant Professor of Biology and Microbiology } Director, Electron Microscopy Facility } Department of Biology and Microbiology } University of Wisconsin Oshkosh } 800 Algoma Blvd } Oshkosh, Wisconsin 54901 } Ph: 920-424-3069 } Fax: 920-424-1101 } E-mail: kostman-at-uwosh.edu } } }
To the person wanting to know if you can keep sections on grids for later labeling: I don't know as I have never tried it. However I have some blocks that are up to 7 years old that from time to time I resection and label with the antibody of the day. In the last few months I have been working with an assortment of Aurion ultra small gold antibodies that can be easily visualized in the scope after silver enhancement. I have been using sections from the same blocks with the same antibodies and I can tell you that using the ultra small gold increases the sensitivity at least ten times over Aurion 10 nm gold particles.
Getting back to your original question, my gut reaction is that there will not be any difference between labeling the sections right away or a week later. Best of luck and let us know what happens with your week old sections, Tim
Timothy Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 238 JAH 1020 Locust Street Philadelphia, Pa 19107 215-513-4798 Work 610-613-8170 Cell
Wrong, the thickness of the glass does not affect the knife-maker, let alone ruin the knife maker.
Understand the workings of that instrument: The weight of the clamping head determines the pressure which is exerted on the glass by the upper two pressure points. The clamping arm locks the head in position, but moving the arm further down does not increase the clamping pressure at all. Clamping pressure could be increased when breaking thicker glass. This is done by adding a kilo or so by pressing onto the head, while locking the clamping head's arm into position.
After clamping and scoring (which is not affected by the clamping of the head, so long as its down) the lower two pressure points are moved up microscopically when rotating the breaker wheel. If thicker glass does not break at this point, you could exert a kilo or two of additional pressure by pushing on the locked clamping head. Although this is very solid and clamped in position, a little extra pressure can hasten the break.
Those knife makers are so solidly made that the additional one or even two kg force I suggest would make no difference.
I expect that the majority of those LKB knife makers in labs today are over 25 years old and they could last another 25 years. Even when used with 8 or 10mm glass. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, August 22, 2001 6:37 AM, Todd Kostman [SMTP:kostman-at-vaxa.cis.uwosh.edu] wrote: } } } Hello all, } } } I have a question regarding breaking glass knives with the LKB 7800 } knife-breaker. When I was trained on the instrument, my advisor threatened } me with instant death if I ever tried to break 8mm thick glass strips rather } than the standard 6.4mm we used. Now I have one of my own and I have a } faculty member who would like to use the thicker glass. Has anyone used 8mm } glass with the 7800 successfully, or was my advisor correct? } } Thanks for your help } } } Todd } } } -- } Dr. Todd A. Kostman } Assistant Professor of Biology and Microbiology } Director, Electron Microscopy Facility } Department of Biology and Microbiology } University of Wisconsin Oshkosh } 800 Algoma Blvd } Oshkosh, Wisconsin 54901 } Ph: 920-424-3069 } Fax: 920-424-1101 } E-mail: kostman-at-uwosh.edu
Think of chromatic aberration as different wavelength light rays (i.e., chromatic) passing through the same point in a lens being focused at different points. CA is a function of wavelength and refractive index of the glass.
Think of spherical aberration as same wavelength light rays (i.e., monochromatic) passing through different points in a lens being focused at different points. SA is a function of the spherical surface of lenses.
It is the challenge of lens design to use different curvature lens surfaces, in conjunction with different refractive index glasses, to bring light waves associated with a point in the object to a focus in the image plane. Planapo objectives are used in conjunction with compensating eyepieces to complete the correction.
Gary Gill
-----Original Message----- } From: Karl Garsha [mailto:keg-at-csd.uwm.edu] Sent: Tuesday, August 21, 2001 12:20 PM To: bigirrell-at-microlinetc.com; Microscopy-at-sparc5.microscopy.com
Just to summarize some of the information regarding NA and Resolution and Magnification: Resolution and numerical aperture are related by resolution = wavelength/N.A. objective + N.A. condenser. Brightness, numerical aperture and magnification are related by brightness= (N.A.)squared /(magnification)squared. As is apparent from these equations, in an idealized situation, a larger numerical aperture at a given magnification will yield better resolution and greater brightness. Things get a little more complicated in real life as has been astutely pointed out, however. Glass has different refractive indexes for different wavelengths of light. White light is composed of red, green and blue wavelengths, and this presents a problem when specimens are illuminated with full spectrum light sources. The red, green and blue components of the image will be refracted by the lens at slightly angles, and so the focal points for the different wavelengths end up being slightly different and the result is that objects appear to be surrounded by a color fringe. This phenomenon, called axial chromatic aberration, is exasperated at greater angles incident to the lens. In other words, at high mag, the higher the numerical aperture the more pronounced the chromatic aberration. It's a concept that is easy to illustrate but hard to explain. The solution, put simply, is to combine different types of glass which have equal but opposite differences in refractive index for light at different wavelengths. This involves more glass, more smart people with handsome salaries to engineer the glass, and results in an expensive plan-apochromatic lens which is corrected for chromatic aberration in the red, green and blue spectra. If the application only requires limited spectra, as in ultraviolet illumination, chromatic aberration isn't such a concern, and one can get by with less expensive lenses of the same numerical aperture, and because of less corrective optics, may indeed be transmit more light to the eye. Spherical aberration is a defect of lenses in which the surface forms part of a sphere. High numerical apertures at high magnification exasperate spherical aberration, so high quality lenses incorporate corrective optics to bring areas on the circumference of the field of view into focal register with the center of the field of view. Again, more glass, more smart people, more money. So the long and the short of it are that high mag lenses don't necessarily have high numerical apertures, however resolving power and brightness will suffer proportionally. High numerical aperture high mag lenses may incorporate artifact into the image unless they are engineered with compensating corrections. Some of these artifacts, which are expensive to correct, may not be an issue under certain circumstances, and may even hinder optical performance, so it is important to understand what corrections one is purchasing, and why.
Karl G.
/**************** Karl Garsha www.uwm.edu/~keg ****************/ ----- Original Message ----- } From: "Bruce Girrell" {bigirrell-at-microlinetc.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, August 20, 2001 11:28 AM
Listers,
Since our switch last year from OEM service contracts to service managed by insurance companies for our EM's, life has been, to put it mildly, interesting. I will be happy to share the details of our experiences with anyone who is having to make this decision (or is just interested), but I didn't want to put a lengthy post on the list if it's not relevant to a fair number of people. Please let me know if you would like to hear our take on this issue.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
You may want to consider a "Polaron" Evaporator, made by Thermo V.G. Scientific in the U.K. They have been manufacturing EM sample prep equipment for many years. Energy Beam Sciences, Inc. is the U.S. agent for Polaron and we would be happy to help you in any way we can.
Best regards,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 mnesta-at-ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: Gillmeister, Russ [mailto:RGillmeister-at-crt.xerox.com] Sent: Tuesday, August 21, 2001 9:37 AM To: 'MSA'
Hi TEM'ers
I need some advice on a new vacuum coater for a TEM lab. I presently have a 13 year old Denton 502A system which has performed satisfactorily. Our main use is for thermal evaporation of Au, C and Al. Denton no longer sells this model but has a new system, an "Explorer 14". I have also gotten a quote for an Edwards "Auto 306" which I know little about. Are there other systems I should be considering? I was considering diffusion systems for reliability. Are turbo systems reliable, and agressive enough now? I would appreciate any comments on the performance of these or other systems. Please respond directly. Thanks for your time. Russ Gillmeister Microscopy Xerox Corp. RGillmeister-at-crt.xerox.com
} I have a question regarding breaking glass knives with the LKB 7800 } knife-breaker. When I was trained on the instrument, my advisor threatened } me with instant death if I ever tried to break 8mm thick glass strips rather } than the standard 6.4mm we used. Now I have one of my own and I have a } faculty member who would like to use the thicker glass. Has anyone used 8mm } glass with the 7800 successfully, or was my advisor correct? } } Todd -
It's possible, but it requires resetting almost everything - which means that it won't break "standard" glass properly at the new settings. This is definitely a "majority rule" situation, unless your faculty member outranks everyone else...
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Dear Gary, We have a Zeiss Axioscope that is setup similar to your axioplan. Over the last five or so years (maybe longer?) it has been well supported by SERCO technical services located in Livermore CA. If you're interested you can contact Emile Meylan or John O'Neill at 1-800-483-0508 they are both very competent. But if you are going to "dump the axioplan" let me know....
Jon Mulholland Botstein Lab, L313 Department of Genetics Stanford University School of Medicine Stanford CA 94305-5210
650-725-1609
On Tue, 21 Aug 2001, Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Jim: } } I am at the point of deciding whether to dump my } Axioplan system for an Olympus BX-51. I'm not sure } that this is a good idea or is a decision in a positive } direction. Thus, I seek your input on this issue. } } As I mentioned before, my current Zeiss system is not working } beyond about 200X. Is there someone in the Sacramento CA } region who can work on my system and find out what is } wrong with it? If some minor tweak is all that is necessary, } I certainly don't need to spend another $40K for a new 'scope. } If I can take the system to someone nearby, that would be } good too. } } I have the basic following items: } } Axioplan 1 stand } 44 72 15 100W lamphouse } 2ea 44 53 65 condenser turrets } 44 52 48 Ph/DIC turret condenser w/ 46 52 68 1.4 top lens } 44 53 51 auto-swingout lens assembly with 0.9NA top lens } 44 53 50 manual swingout lens assembly with 0.9NA & 1.4 top lenses } } Plan Neofluar 1.25/0.075 } Plan Neofluar 20X/0.5 Ph2 } Acrostigmat 100X/1.25 Ph3 } Plan Neofluar 40X/0.75 Ph2 } Plan Neofluar 63X/1.25 oil } Plan Neofluar 20X/0.5 with DIC prism } Plan Neofluar 40X/0.75 with DIC prism } Plan Neofluar 40X/1.30 oil } Plan Neofluar 5X/0.15 } Plan Neofluar 10X/0.30 } Plan Neofluar 63X/1.25 oil with DIC prism } Plan Neofluar 100X/1.30 oil with DIC prism 44 4 80 } } 44 52 11 swingout low power condenser } 44 53 12 thing-a-ma-bob } } 45 29 21 stress-free trinoc head } 43 36 05 analyzer } Polarizer } Ph 1,2,3 inserts } DIC 1,2,3 inserts } 2ea 45 31 80 DIC nosepieces } } Is there any chance for support and/or parts for this system? } Is it a lost cause? You indicated that these systems were } still supported. How can I get such support? } } gary gaugler } } } } At 03:58 PM 7/25/2001, you wrote: } } } Dear Dr. Gaugler, } } } I want to thank you for taking the time to talk to me yesterday. I appreciate } } your honesty and frankness. We are always glad to hear from our customers, } } especially if the feedback is constructive. Certainly, I was pleased to hear } } your praise for the performance our Microscopes deliver. Of course, I was } } surprised to hear of your negative experience with our support } } team. Therefore, } } I would like to answer your email and phone conversation in two parts: } } [snip] } }
Copies can be made available. Responses/requests OFFLINE only please.
Regards,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 CASI Home Page: email: fmonson-at-wcupa.edu CASI Home Page: http://darwin.wcupa.edu/casi/ Schedule: Navigate from CASI Home Page Please call before visiting.
We have an Oxford ISIS EDS system. As it stands it is not set up to acquire spectral images. We would like to be able to do spectral image acquisition on this machine. Is there anyone out there who knows how the system may be modified (macros, interfaces, whatever) to do it? We do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that helps.
-- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
If those fail, call me at 916.791.8191 or e-mail and I can probably find one locally.
gary
At 09:56 AM 8/22/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Randy, I, for one, would be most interested in your experiences and possibly comparison with your OEM experiences. As a third party service company I would find the feedback helpful in tweaking my own contracts to better serve the users. All three modes have pluses and minuses for a number of valid reasons. How various customers view those +'s, -'s and reasons for them is helpful to a lot of us on both sides of the fence.
Ken Converse owner Quality Images third party SEM service Delta, PA 17314
Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listers, } } Since our switch last year from OEM service contracts to service managed by } insurance companies for our EM's, life has been, to put it mildly, } interesting. I will be happy to share the details of our experiences with } anyone who is having to make this decision (or is just interested), but I } didn't want to put a lengthy post on the list if it's not relevant to a fair } number of people. Please let me know if you would like to hear our take on } this issue. } } Cheers, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } }
If you have a vacant position for a Ph.D holder in Materials Science and Engineering (Electron microscopy/Materials characterization) that needs to be filled immediately, please contact me at benedict-at-u.arizona.edu.
We supply Leit-C in Oz, though I have not yet listed the material in our online. I guess its true for many suppliers: however large our online catalogue is we have ready access to two or three times as many items. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Wednesday, August 22, 2001 2:51 PM, Cheng Huang [SMTP:HUANG-at-rsbs.anu.edu.au] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } We would like to order some conductive carbon cement (Leit-c), but were told } that we have to pay more than $100 for special package over the price of the } cement because of the chemical contents in the cement. We wonder who has } bought Leit-C recently and through which local dealer in Australia. } Thanks for any information. } Cheng Huang } } ----------------------------------------------------- } Cheng Huang } Australian National University } EM Unit, RSBS } Box 475, ACT 2601 } Canberra, Australia } Phone: 61-2- 6125-6553 } Fax: 61-2- 6125-3218 } http://www.anu.edu.au/EMU/ }
By spectral images, do you mean xray mapping? A "key" disk for the ISIS software program is required to activate the xray mapping facility. Of, course this will cost a bit of money to purchase from Oxford. When the ISIS program is loaded initially, all the software required to use the program is there, but activating the unpaid for parts of the program is the thing.
Fred
On Wed, 22 Aug 2001, Alwyn Eades wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } We have an Oxford ISIS EDS system. As it stands it is not set up to } acquire spectral images. We would like to be able to do spectral image } acquisition on this machine. Is there anyone out there who knows how } the system may be modified (macros, interfaces, whatever) to do it? We } do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that } helps. } } -- } .......... } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvania 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu } }
I would be interested. I have never had a "good" experience with any claim from any insurance company.
The money is the only thing that matters with them.
Regards,
Earl Weltmer
Claimer (not DISclaimer) : The above opinion are my own and are necessarily the opinion of my company. They are based upon actual experiences with insurance comapnies over the years from health insurance (Blue Cross), to auto (Farmer's), to moving Companies (United Van lines, FEDEX).
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 22, 2001 8:35 AM
Hi Alwyn,
This is something I thought about a while back. Unfortunately, I didn't come up with any ideal solutions. You can define an 2-D array of points using the AUTO module and an appropriately short value of live time. This will save a full spectrum for each point on disk in separate sequentially numbered files (painfully slow and get a larger disk). I don't remember what the limits on the dimensions of the array are. The big problem is then how to analyze the data. Within the ISIS software the only thing I can think of is to batch process for peak integrals. Ray Twesten has written a short utility which can be used to batch convert the ISIS format spectra to 1-column ASCII format. From here you could write your own processing routines or perhaps use digital micrograph (or a little of both). I'm sure many of us would be intested if you find a workable solution?
P.S. I just remembered Oxford also had a little known programming API product, which could be used to control and read from the ISIS hardware from your own application. I forget what they called it. I suspect this would be a major undertaking to work with.
Jim Mabon
-----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] Sent: Wednesday, August 22, 2001 1:22 PM To: EMNET
We have an Oxford ISIS EDS system. As it stands it is not set up to acquire spectral images. We would like to be able to do spectral image acquisition on this machine. Is there anyone out there who knows how the system may be modified (macros, interfaces, whatever) to do it? We do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that helps.
-- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I beg to differ and stand by my previous contribution, which is appended last. Caroline talks about changing settings. Well, the settings adjust the lateral position of the score mark and how far forward the score is to extend. These adjustments need to be changed when cutting strips other then squares (25.4mm), or for non 45 degree knives. Drawing a felt tip marker across the correct settings (different colours for different settings) makes it easy to find the correct settings again. Anyway, those settings have nothing to do with the thickness of the glass. Similarly, the clamping pressure is gravity (weight of clamping head) and the head will exert the same pressure regardless of the glass thickness. Dto the score pressure.
Fred is concerned about half inch glass, but the original question concerned 8mm glass. I have no hesitation to use (and have) 8mm glass with the LKB knifemaker. The 1/4 inch (6.4mm) thickness glass has been a convenient standard material for EM, but the design is not specific to that thickness. I expect that 10mm glass would also be no problem with the LKB, half inch (12.6mm) would require still greater pressure, and perhaps there is a limit, but I cannot see that reasonably any harm could be done to those very solidly build part of the knifemaker.
Incidentally, years ago LKB was the sole agent for "LKB" (actually Alkar) glass. That glass is made up to 10mm thick and I cannot imagine that LKB marketed that glass for use with another knifemaker. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
-----Original Message----- } From: Caroline Schooley [SMTP:schooley-at-mcn.org] Sent: Thursday, August 23, 2001 1:51 AM To: Todd Kostman Cc: Microscopy-at-sparc5.microscopy.com
Dear Earl and all else,
You may think this sounds so extreme that it is beyond belief. Sad to say that not everyone has sound judgement and have fallen for this ruse. At worst, the victims are eventually duped to travel to Nigeria and held for ransom. Some have been killed.
No substitute for earning a living the old fashioned way.
Al Stone
At 07:24 AM 8/18/2001 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } 1) tighten the block in the chuck } 2) tighten the knife in its holder } 3) check knife angle(I use 4 deg. clearance angle) } 4) try at different times of the day- at my old place my lab was next to } two large turbines and early in the day I occasionally had problems with } chatter } }
Here are a few more suggestions:
1) Use distilled water in your boat as opposed to de-ionized. 2) If possible re-trim your block. Even though you think things look good, sometimes just cleaning up the edges with a new razor blade makes things better. 3) As much as you can, keep your block chuck and knife stage centered on zero. It seems for me if I swing the knife to the left or right too far, chatter is a given. 4) Try another knife. I've had diamonds re-sharpened and I swear they came back duller than they started out. Also something could have happened to the mount and the diamond may be loose. 5) Pray to the Microscopy gods. :-)
Hope this helps,
Paula Moore Wake Forest Univ. Medical Center Pathology/EM Lab
Problem solved! thanks to Bill Sharp! As always the Microscopy list is the best! thanks for all the help, Beth
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
As promised, here is a rundown on our OEM vs. insurance experiences. I decided to put this on the list based on the very large number of replies I received, some asking specifically that this be posted. As a result, I've removed all names, since I don't have a clue as to legal ramifications of being more specific. Sorry to be so wimpy, but in these cases I believe in CYA.
The following obviously reflects our experiences alone, but based upon what I have heard from a few others, I don't believe that our situation is unique, by any means.
Our lab, like many others, has been under increasing pressure to cut costs for the last two to three years, and since service contracts represent a significant proportion of our budget, they logically became a cost-cutting target immediately. It was beginning to appear that we were going to lose the contract on one of our TEM's to save money, especially since it wasn't heavily used. About this time one insurance company (IC, for short) began a series of high-powered presentations to the University purchasing people and administrators, promising significant cost reduction, with no reduction in services, choice of service providers, elimination of paperwork, etc. Facilities like ours were contacted by University purchasing staff in order to set up individual meetings to assess our needs and get estimates on the IC's costs versus OEM charges.
After attending two open seminars and having two meetings with the insurance people and purchasing staff in our facility, the decision was made, with my agreement, to give the IC a shot at managing our service. The thinking was that we would try it for a year. The IC said we could back out at any time with no penalty, they would pro-rate our contracts to adjust for the differing ending dates for our OEM contracts, and, they said, there should be no "re-certification fee" if we decided to go back to OEM contracts, since we intended to use only the OEMs' engineers anyway. They would take care of the paperwork notifying the OEMs about the new arrangement, and all billing would be handled between the IC and the service providers. They would guarantee no increase in contract costs for, I believe, three years.
The very first service call we made to the manufacturer on one of our TEMs resulted in our being told that they couldn't send an engineer, because we weren't on service contract any longer. When I explained that they were supposed to have been notified that we were with the IC, I was told that we would have to provide a University P.O. before anyone would be sent. Apparently, there was a substantial outstanding bill from this IC for service provided to a university in another state, and until it was paid no service would be provided on our contract. The IC's on-campus rep, our purchasing director, and the director of the university core facilities were notified by me immediately. Apparently some high level phone calling went on, because before the day was out, the service provider had called back saying the outstanding bill would be paid and an engineer was on the way. (Additionally, our purchasing director assured us that we could get a P.O. if necessary. We have terrific support here from purchasing and they have been invaluable on several occasions.)
The engineer was sent out, the scope was back on line. BUT, it turns out that the promised payment was never made and we were back to square one the next time we needed service on that machine. In fact, it was worse, because the OEM was never paid for the service on our scope either.
Our other scopes are from a different manufacturer and initially service was pretty much the same as before (although they made it clear they weren't happy about the switch). Response time was a bit slower, and there was more stinginess with replacement parts, but we had no real complaints.
Then, the IC declared bankruptcy. Their campus reps disappeared, their web site went down, and they effectively vanished. We had already used part of the year's contract, but were only able to recover about $700 on the unused portion. Some labs lost thousands, I've been told. The service provider was never paid, and lost a substantial amount of money. (I'm told that this IC has reorganized under a different name and is soliciting new business, especially from universities. Ask lots of questions if a new company comes around. Be especially careful of large numbers of enthusiastic people in nice suits giving slick multi-media presentations!)
We began discussing returning to our old OEM contracts and I was asked to get prices. The first thing I was told by one service provider was that we would have to have our instrument re-certified, which would cost about $1500, even though nobody had touched that scope but the OEM engineers. Needless to say, that rankled a bit, so we checked into another insurance company that had been recommended by several other labs. We were immediately impressed by the fact that they presented what seemed to be a more realistic picture of what they could do for us. They also offered significant savings (10-15%) over the old contracts, so we once again decided to give it a try.
This time, the first call we made had the same result as before, i.e., the provider wanted a P.O. before an engineer would come out. When I asked why, they said that even though they had no problems with the new IC, they weren't going to take a chance on losing a ton of money again. If the university would sign a waiver accepting responsibility for paying any amounts in default, we could get service again. Fine, I said. Several days later the form was faxed to us and I forwarded it to our purchasing director, who responded that he had passed it along for review to the people who needed to sign off on it. That was last month sometime, and I have heard nothing yet. So far we have been waiting to get a preventive maintenance visit on this scope for about two months and nothing's in sight, yet.
On our other scopes, we have again been able to get service, but now it seems that we are way, way down on the priority list. OEMs will tell you that they are obligated to service their contract customers first, and in our experience they certainly do. The service management (insurance) companies will tell you that this is not the case, that it's first-come, first-served. That has not been our experience. On one TEM we have been waiting weeks for a PM. It was scheduled twice and an engineer was here, but was pulled away on "emergency" calls, which I read as calls from contract holders. The PM has not yet been done. (Just minutes ago we received a call from our service engineer saying he's been pre-empted again and now he has NO idea when he can get here.) Our SEM has been serviced under the new arrangement in a relatively timely manner.
Under the OEM contracts, service was provided "yesterday" and parts were replaced if they were even suspected to be bad. The working relationship was wonderful, and the service was stellar. We were called and reminded of PMs before they were due and we received periodic phone calls just to check on the status of our equipment. Under the new contracts, service is provided (if we can get it) whenever they get around to it, and engineers can be interrupted by calls to respond to contract holders. Parts are changed less frequently and are all billed. We schedule the PMs (no big deal). The relationship is much more tense than it used to be.
We could probably increase the level of service by reminding the service people that when it's time to replace our scopes, service history will have a MAJOR bearing on whose machines we buy. However, I'm extremely reluctant to turn a previously very pleasant relationship into a confrontational, grudging one (although it seems to be headed that way, anyway). In addition, I can understand why some of these things are happening. If I put myself in the shoes of the OEM service managers, I'm not sure what I would do differently. On the other hand, I also have a tiny, nagging feeling that part of these problems are deliberately designed to pressure us back into our old contracts. That possibility (and I don't know it to be true) annoys me intensely, but I still haven't played the sales managers against service managers card. My sense is that OEMs make big bucks on service contracts and they take it VERY personally when we "fire them" as one service manager phrased it.
I would like to emphasize very strongly that when field service engineers come out, they are uniformly excellent. They do everything they are allowed to do to provide top-notch care for the instruments. The problems originate higher up.
In summary, we have found that service management companies do indeed save money on comparable contracts, but we have had a miserable time getting service at all when using them. This may not be typical---I don't know. Our level of service has declined greatly in terms of promptness, attitude, and parts supply. The attitude part was certainly not worth the extra several thousand dollars the OEMs wanted, but the promptness and parts issues probably were. Most disturbing was finding out that we could be denied service because of a problem originating at another university in another state that had absolutely nothing to do with us.
We will continue for now with our new IC's to see if the problems correct themselves, especially since the problems didn't arise with this company. In addition, one of the OEM service managers called and suggested we might try a custom contract in which we could work with them to design our own service schedule. He said we could be informally trained during service visits to perform many of the maintenance chores on the scope and could retain a contract that would basically cover emergencies, at a substantial cost savings. We are looking into this now, because it seems to have lots of nice possibilities. (We already do filament/gun exchanges and cleanings, obviously, but I've never personally done a mechanical column alignment or routine maintenance requiring major disassembly.)
My best advice: if you are currently with an OEM, stay there. If you are losing your local in-house service wizard and need to decide on contracts, go with the OEMs. Fight like hell to resist pressure to switch.
If you are with an insurance company and are happy with them, stay there, and I wish you continued good luck. Please tell me your secret.
My suggestion to OEM service providers: offer some in-house maintenance training courses, better maintenance manuals and schematics, and encourage customers to do more maintenance tasks (with appropriate provisos for covering dumb mistakes). Lower your contract prices so we're not so pressured to drop them. The glory days are over, folks. Time to compete.
As for me, I have no financial interest in any of these folks. I just want our scopes to work. Feel free to contact me with any questions.
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
This is correct, the keydisk is all that is required for the software to be enabled. However, there may be hardware required, too, at least a cable interface (RS232?) to the microscope. I believe the ISIS Autobeam likes to select its own scan rates. You may need to speak with Oxford re: compatibility with your particular microscope.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Wednesday, August 22, 2001 9:23 PM, Fred Pearson [SMTP:eoptics-at-mcmail.cis.mcmaster.ca] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Alwyn: } } By spectral images, do you mean xray mapping? A "key" disk } for the ISIS } software program is required to activate the xray mapping } facility. Of, } course this will cost a bit of money to purchase from } Oxford. When the } ISIS program is loaded initially, all the software } required to use the } program is there, but activating the unpaid for parts of } the program is } the thing. } } Fred } } } On Wed, 22 Aug 2001, Alwyn Eades wrote: } } } } } --------------------------------------------------------- } } --------------- } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } ml } } } } --------------------------------------------------------- } } --------------. } } } } } } } } We have an Oxford ISIS EDS system. As it stands it is } } not set up to } } acquire spectral images. We would like to be able to do } } spectral image } } acquisition on this machine. Is there anyone out there } } who knows how } } the system may be modified (macros, interfaces, } } whatever) to do it? We } } do have Digital Micrograph (on a Mac whereas ISIS is on } } a PC), if that } } helps. } } } } -- } } .......... } } Alwyn Eades } } Department of Materials Science and Engineering } } Lehigh University } } 5 East Packer Avenue } } Bethlehem } } Pennsylvania 18015-3195 } } Phone 610 758 4231 } } Fax 610 758 4244 } } jae5-at-lehigh.edu } } } } } }
We are looking to acquire a Link AN10000 EDS system for a Hitachi H-600 TEM. Currently this EDS system is on a H-7000 STEM. My questions is: will an EDS system that fits the H-7000 STEM, also fit the H-600 microscope?
Also, please share your experiences with the Link AN10000 EDS system - its reliability, support, etc.
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I am pretty sure that Alwyn is not talking about x-ray mapping. If that was all he wanted, I suppose the key disk is all that would be needed if he already has the other imaging applications and hardware installed.
Someone suggested using Auto to scan an image for multiple spectra. Indeed, the software and hardware should allow it. The spectra might then be exported and processed by some other package to perform the type of component analysis that has been described at MSA over the last few years. My current ISIS x-ray application (version 3.32) has the option of storing the x-ray data in single column MSA format. Previous versions placed multiple channels on a single line.
One hitch would be the limitation on the number of spectra per job. I think there is a limit of 1000 or so built into the database used for managing the data. I suppose it could be changed to another number, but we bumped into it some time back.
That would mean you could do a 32x32 raster of points saving a spectrum at each. A four second acquisition per point would mean a bit more than an hour for acquisition. That would not be too bad, but the spatial resolution would probably be too low to be very useful. But the data should readily fit onto hard drives with capacities of a few gigabytes.
I would be interested in seeing how the data processing is handled. If someone makes it work, I would be very interested in hearing the details.
Warren
At 10:33 AM 8/23/2001 -0400, you wrote: } -----------------------------------------------------------------------. } This is correct, the keydisk is all that is required for } the software to be enabled. However, there may be hardware } required, too, at least a cable interface (RS232?) to the } microscope. I believe the ISIS Autobeam likes to select } its own scan rates. You may need to speak with Oxford re: } compatibility with your particular microscope. } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Wednesday, August 22, 2001 9:23 PM, Fred Pearson } [SMTP:eoptics-at-mcmail.cis.mcmaster.ca] wrote: } } } } Alwyn: } } } } By spectral images, do you mean xray mapping? A "key" disk for the ISIS } } software program is required to activate the xray mapping facility. Of, } } course this will cost a bit of money to purchase from Oxford. When the } } ISIS program is loaded initially, all the software required to use the } } program is there, but activating the unpaid for parts of the program is } } the thing. } } } } Fred } } } } On Wed, 22 Aug 2001, Alwyn Eades wrote: } } } We have an Oxford ISIS EDS system. As it stands it is not set up to } } } acquire spectral images. We would like to be able to do spectral image } } } acquisition on this machine. Is there anyone out there who knows how } } } the system may be modified (macros, interfaces, whatever) to do it? We } } } do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that } } } helps. } } } .......... } } } Alwyn Eades } } } Department of Materials Science and Engineering } } } Lehigh University } } } 5 East Packer Avenue } } } Bethlehem } } } Pennsylvania 18015-3195 } } } Phone 610 758 4231 } } } Fax 610 758 4244 } } } jae5-at-lehigh.edu
I am investigating EDS systems to integrate with a JEOL-6100 SEM. I'm leaning towards PGT's Spirit system. Other considerations are Oxford's INCA and Thermo Noran's Vantage. Does anyone have particularly positive or negative experience with PGT? I don't often hear much about that company. Any comments about the other possibilities would be appreciated, as well.
Thanks!
-Peggy
Peggy McKarns Macatangay, PhD Project Analyst 2 Celanese Corpus Christi Technical Center
Has anyone seen or heard from John Best at the ELMDAS Company? They/He manufacture the "DIGISEM" SEM Digital Imaging System. I along with many other people(customers) have left many messages(voice & email) to no avail. Any help would be greatly appreciated. Please reply directly to me. Thanks in advance.
Gary M. Easton Scanners Corporation {mailto:gary.easton-at-scannerscorp.com} gary.easton-at-scannerscorp.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (john_bruss-at-bose.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 22, 2001 at 17:00:52 ---------------------------------------------------------------------------
Email: john_bruss-at-bose.com Name: John Bruss
Organization: Bose Corp.
Education: Graduate College
Location: San Diego, CA, USA
Question: What materials and process would an amateur use to suspend a spider in a clear cube for unmagnified artistic and historic use? Where are those materials available in small quantities?
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Hi all, This post is directed primarily to those doing biological prep in the midwest. I am sure that a number of you were as impressed with the recent advances in microwave specimen prep presented at M&M2001 as I was. I have threatened in the past to have a workshop at Purdue so we could get up to speed on these new techniques.
Well the time has come.....or at least will come if there is sufficient interest. The proposed hands-on workshop, conducted by Rick Giberson of Ted Pella, Inc., would be 2-2 1/2 days in length. Exact cost would not be known until we have an idea of numbers of participants. The dates for the workshop will be set after determining interest but hopefully will be within the next few month.
Available would be two microwaves, 2 TEM's and 1 SEM to check samples, 5 microtomes, misc. other equipment, etc so that all attending would be able to process and check samples.
Purdue is located in West lafayette, Indiana which is ~ 1hr. north of Indianapolis or 2hr SE of Chicago by car. It is serviced by Northwest airlines via the Lafayette/Purdue airport or via the airport in Indianapolis.
Please respond immediately if you are seriously interested in attending a workshop. I will be in touch with more details to those responding as soon as we determine whether the workshop will be held.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id OAA18359 for dist-Microscopy; Thu, 23 Aug 2001 14:17:14 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id OAA18356 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 23 Aug 2001 14:16:43 -0500 (CDT) Received: from es05.hosts.jhmi.edu (es05.hosts.jhmi.edu [162.129.224.75]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id OAA18349 for {microscopy-at-sparc5.microscopy.com} ; Thu, 23 Aug 2001 14:16:31 -0500 (CDT) Received: from welchlink (welchlink2 [162.129.224.74]) by es05.hosts.jhmi.edu (8.9.3/8.9.3) with ESMTP id PAA29299 for {microscopy-at-sparc5.microscopy.com} ; Thu, 23 Aug 2001 15:10:49 -0400 (EDT)
Hi, everyone, Does anyone have the recipe and protocol on how to make uranyl formate solution for negative staining? And do you have any idea about whhich is better, uranyl aceate or formate? Thanks a lot.
Chen Chen
Dept of Biol. Chem. the Johns Hopkins Univ. School of Medicine
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, August 23, 2001 7:23 AM
A lab I am working in for a while has a new Nikon Eclipse inverted and when I went to use it, I could not get the DIC to work. The rep came by today and fixed it by turning over the slider holding what I believe is the initial polarizer (the one above the body that presumably is the first polarizer in the transmitted light path). I have not seen a real detailed description of the Nikon DIC so I do not understand what each element is and why this should have had so great an effect. I thought the rotatable element below the condenser was a rotatable polarizer, so I thought at some point I would have gotten extinction even if the orientation of the first was changed. Thanks- Dave
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mcs4-at-dana.ucc.nau.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, August 23, 2001 at 11:04:58 ---------------------------------------------------------------------------
Email: mcs4-at-dana.ucc.nau.edu Name: Matthew Salanga
Organization: Northern Arizona University
Education: Graduate College
Location: Flagstaff, Arizona
Question: I have been doing TEM work on plant cells for my thesis. I was wondering if anyone might have a suggestion on a stain which would be specific for beta glucans mainly callose. Literature suggests that normal UA and lead citrate do not stain callose at all, however it appears as though it may according to what I believe is Callose in my micrographs. Anyways if anyone has suggestions or comments please email me.
Sorry to use the list for this, but my email to Tom Gore got bounced. Tom -- your IT department doesn't believe you exist. I got this error message when I emailed you: The original message was received at Thu, 23 Aug 2001 16:49:49 -0500 from [144.92.45.161]
----- The following addresses had permanent fatal errors ----- {togo-at-uvvm.uvic.ca}
----- Transcript of session follows ----- .. while talking to smtp.uvic.ca.: } } } RCPT To: {togo-at-uvvm.uvic.ca} { { { 553 5.3.0 {togo-at-uvvm.uvic.ca} ... Mail from 144.92.9.40 rejected(outputs);see http://orbz.org 550 {togo-at-uvvm.uvic.ca} ... User unknown
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
We've had the same budgetary problems with oem contract maintenance on our Philips CM120. We could only do 3 years of a 5 year contract, renewable every year. At $16,000 to $18,000 for the fourth and fifth years, our Dean bulked at the cost for what is essentially an insurance policy, and so we didn't renew for the last two years. Our choice was to go with an independent contractor, who formerly worked for the OEM, and got good service. We still couldn't do a service contract with him (less than half the OEM) because our funding is very limited. Instead we used an open or blanket $2500 service order. He at times bent over backwards to help us, but without a service contract, he could not be "on-call." Another limitation was a need for proprietary diagnostic software that he no longer had access to. We had to go back to the OEM for a one-time service.
The upshot is that you should try an independent service. Their requirements will differ, but your alternative is to pony up $10-20 grand for "insurance". Maintenance on new microscopes that are computer controlled maybe problematical for in-house training especially when proprietary diagnostic software is needed. If buying a new microscope, get as much funding for 5 years of service as possible. We needed the 2 year warranty plus the 3 year service contract before it performed steadily without problems. We're starting into early middle age on our CM120 and experiencing increasing problems. Our first TEM bought in 1962 lasted 30 years and was used for teaching EM and research. We were able to operate it without a service contract. I think we'll never see that kind of performance again, but then they were much simplier machines.
-- Greg Lum Lab Manager Electron Microscope Facility San Francisco State University College of Science & Engineering Ph: 415/338-1339 Email: glum-at-sfsu.edu
I'd like to respond from the viewpoint of one who worked for an OEM for 4 years and has subsequently had his own third party service company for 20 years.
First, the OEM is obligated to service contract customers first. You have become a "billable" call and DO go to the bottom of the list. That would not be all that different with a third party firm (although they might try a little harder to be timely). The contract people have paid up front (or have at least committed up front to pay) for an entire year. This is guaranteed income and includes a guaranteed liability (for the OEM) "to maintain the instrument/system to its original specifications". Service organizations have to take this seriously. This liability doesn't exist for billable customers. Work will be done to the best of the OEM's ability and not beyond the limit of the purchase order.
Second, the IC has no technical expertise, no intimate connection with its customers and no obligation to use the manufacturer. I have been called by at least one IC several times, but have never done any work for them because they haven't a clue what I'm talking about when I tell them I service SEMs but not TEMs and I like to stay within 500 miles of home. (Their geography hasn't impressed me, either). They are trying to emulate what happens when your car gets in a fender bender. There are lots of people who can fix it, and many of them can do a pretty satisfactory job. But ask yourself, "When my car has a rumble, or the engine misfires and I need expert help or maintenance, who do I call? My insurance company?" Probably not, not only because your insurance doesn't cover that kind of thing, but because they couldn't help you anyways. They know about insurance, not the inner workings of your car.
Third, any outside party that starts making commitments FOR the OEM (you won't have to pay for recertification..............) I would turn away from and run, not walk, run. If they tell you that THEY will pay for recertification if you're unhappy and they will put it in writing, that's another story.
As to the manufacturers making lots of money on their contracts, some may, some may not. A lot depends on how well they are set up and run. It also depends on the instrument density. If one engineer can service 20 or 30 systems within driving distance, the engineer is competent and the service department backs him up well, yes, it can be quite profitable. If you have to fly to every site, you're short on experienced engineers and you don't support them well, you can lose your shirt ( and have a whole lot of PO'd customers to boot).
Every time I've gotten one of those glib calls from an IC in Texas, in the back of my mind a little voice is always saying "And just what makes you think you're going to get paid?" I do thank you for confirming that this concern is warranted.
The question now comes down to "What do you want?". The OEM has reason to feel miffed. You want their expertise, their parts, their top level response, their best guess on advance parts replacement for reliability and you're going to pay someone ELSE the bonus if they do all this. The IC is betting that the instrument will run reliably as it always has, meaning very little cash out. The OEM no longer has the incentive to go out of its way to insure reliability. They'll actually make more if the system breaks more frequently. And as I pointed out, if they do their best work, you pay someone else the bonus while they still have the considerable expense of running a service department. I'm not implying in any way that they will go out of their way to make it fail. One of the advantages of sticking with the organization that actually does the servicing is that you establish a raport with the personnel in the field and in house. I'm only saying that they aren't going to go out of their way to KEEP it from failing, because it's not in their interest to do that. It IS in their interest when they have guaranteed the performance for a full year.
Yes, generally speaking my service contracts are profitable, but there is another major reason that I prefer to have systems on contract. It becomes up to me how much time I spend on an instrument and how fast or slow I work on it. Sometimes I'm very sure what the problem is and fix it immediately, but often there are intermittent problems and I can tell you that a billable customer watches the clock like a hawk when he sees me sitting in front of a running system waiting for an intermittent problem to show up. Sometimes there are subtle problems that the customer isn't even aware of, yet. If the system is on contract I can work without watching the clock and get everything right. In the long run it saves me repeat trips. Also, if I should decide to let something slide due to whatever circumstances, I know that I'm guaranteeing the work and a later failure will be fixed at MY expense, not the customer's.
There was a time when people seemed more likely to reward good service by paying a premium for it and showing a little loyalty to those who provided it. The loyalty seems to be going by the board, and more and more people want more for less, they aren't willing to pay for quality because in their everyday life they buy cheap and when it breaks they throw it out and buy cheap again. I don't believe that the scientific instrument market will get there any time soon. This is specialty equipment, capital investment, there for the long haul.. If it's well taken care of , it will last for decades. That's where you can get the savings.
If you were happy with the quality of the OEM's service and they are willing to work with you, by all means see what you can work out! If you weren't so happy with an OEM, see if there is a third party service company that you can establish a good relationship with. The ICs can't charge less, take out a middleman's cut, and give the organizations who actually do the work and support the equipment, enough money to maintain the quality you demand. If it looks too good to be true...................................................
I sympathize with the cost-cutting that is being forced on you from the bean-counters, but beware of false economies. Maybe one or more instruments could be taken off contract, but what kind of grants are you (the school and faculty) going to lose if it's down too much of the time? What are the priorities of the organization? University service labs are not supposed to be profit centers. They are SERVICE labs and are heavily subsidized because the typical student (grad or undergrad) can't afford to pay commercial rates for instrument time (especially with what they're paying for tuition). If these instruments are important to various departments then the decision is too important to be made by some MBA who doesn't know the difference between hydrogen embrittlement and cilia.
It would be interesting if we could get a response from the insurance industry side of things.
My $.02 worth.
Sincerely,
Ken Converse owner Quality Images third party SEM service Delta, PA
Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } As promised, here is a rundown on our OEM vs. insurance experiences. I } decided to put this on the list based on the very large number of replies I } received, some asking specifically that this be posted. As a result, I've } removed all names, since I don't have a clue as to legal ramifications of } being more specific. Sorry to be so wimpy, but in these cases I believe in } CYA. } } The following obviously reflects our experiences alone, but based upon what } I have heard from a few others, I don't believe that our situation is } unique, by any means. } } Our lab, like many others, has been under increasing pressure to cut costs } for the last two to three years, and since service contracts represent a } significant proportion of our budget, they logically became a cost-cutting } target immediately. It was beginning to appear that we were going to lose } the contract on one of our TEM's to save money, especially since it wasn't } heavily used. About this time one insurance company (IC, for short) began a } series of high-powered presentations to the University purchasing people and } administrators, promising significant cost reduction, with no reduction in } services, choice of service providers, elimination of paperwork, etc. } Facilities like ours were contacted by University purchasing staff in order } to set up individual meetings to assess our needs and get estimates on the } IC's costs versus OEM charges. } } After attending two open seminars and having two meetings with the insurance } people and purchasing staff in our facility, the decision was made, with my } agreement, to give the IC a shot at managing our service. The thinking was } that we would try it for a year. The IC said we could back out at any time } with no penalty, they would pro-rate our contracts to adjust for the } differing ending dates for our OEM contracts, and, they said, there should } be no "re-certification fee" if we decided to go back to OEM contracts, } since we intended to use only the OEMs' engineers anyway. They would take } care of the paperwork notifying the OEMs about the new arrangement, and all } billing would be handled between the IC and the service providers. They } would guarantee no increase in contract costs for, I believe, three years. } } The very first service call we made to the manufacturer on one of our TEMs } resulted in our being told that they couldn't send an engineer, because we } weren't on service contract any longer. When I explained that they were } supposed to have been notified that we were with the IC, I was told that we } would have to provide a University P.O. before anyone would be sent. } Apparently, there was a substantial outstanding bill from this IC for } service provided to a university in another state, and until it was paid no } service would be provided on our contract. The IC's on-campus rep, our } purchasing director, and the director of the university core facilities were } notified by me immediately. Apparently some high level phone calling went } on, because before the day was out, the service provider had called back } saying the outstanding bill would be paid and an engineer was on the way. } (Additionally, our purchasing director assured us that we could get a P.O. } if necessary. We have terrific support here from purchasing and they have } been invaluable on several occasions.) } } The engineer was sent out, the scope was back on line. BUT, it turns out } that the promised payment was never made and we were back to square one the } next time we needed service on that machine. In fact, it was worse, because } the OEM was never paid for the service on our scope either. } } Our other scopes are from a different manufacturer and initially service was } pretty much the same as before (although they made it clear they weren't } happy about the switch). Response time was a bit slower, and there was more } stinginess with replacement parts, but we had no real complaints. } } Then, the IC declared bankruptcy. Their campus reps disappeared, their web } site went down, and they effectively vanished. We had already used part of } the year's contract, but were only able to recover about $700 on the unused } portion. Some labs lost thousands, I've been told. The service provider } was never paid, and lost a substantial amount of money. (I'm told that this } IC has reorganized under a different name and is soliciting new business, } especially from universities. Ask lots of questions if a new company comes } around. Be especially careful of large numbers of enthusiastic people in } nice suits giving slick multi-media presentations!) } } We began discussing returning to our old OEM contracts and I was asked to } get prices. The first thing I was told by one service provider was that we } would have to have our instrument re-certified, which would cost about } $1500, even though nobody had touched that scope but the OEM engineers. } Needless to say, that rankled a bit, so we checked into another insurance } company that had been recommended by several other labs. We were } immediately impressed by the fact that they presented what seemed to be a } more realistic picture of what they could do for us. They also offered } significant savings (10-15%) over the old contracts, so we once again } decided to give it a try. } } This time, the first call we made had the same result as before, i.e., the } provider wanted a P.O. before an engineer would come out. When I asked why, } they said that even though they had no problems with the new IC, they } weren't going to take a chance on losing a ton of money again. If the } university would sign a waiver accepting responsibility for paying any } amounts in default, we could get service again. Fine, I said. Several } days later the form was faxed to us and I forwarded it to our purchasing } director, who responded that he had passed it along for review to the people } who needed to sign off on it. That was last month sometime, and I have } heard nothing yet. So far we have been waiting to get a preventive } maintenance visit on this scope for about two months and nothing's in sight, } yet. } } On our other scopes, we have again been able to get service, but now it } seems that we are way, way down on the priority list. OEMs will tell you } that they are obligated to service their contract customers first, and in } our experience they certainly do. The service management (insurance) } companies will tell you that this is not the case, that it's first-come, } first-served. That has not been our experience. On one TEM we have been } waiting weeks for a PM. It was scheduled twice and an engineer was here, } but was pulled away on "emergency" calls, which I read as calls from } contract holders. The PM has not yet been done. (Just minutes ago we } received a call from our service engineer saying he's been pre-empted again } and now he has NO idea when he can get here.) Our SEM has been serviced } under the new arrangement in a relatively timely manner. } } Under the OEM contracts, service was provided "yesterday" and parts were } replaced if they were even suspected to be bad. The working relationship } was wonderful, and the service was stellar. We were called and reminded of } PMs before they were due and we received periodic phone calls just to check } on the status of our equipment. Under the new contracts, service is } provided (if we can get it) whenever they get around to it, and engineers } can be interrupted by calls to respond to contract holders. Parts are } changed less frequently and are all billed. We schedule the PMs (no big } deal). The relationship is much more tense than it used to be. } } We could probably increase the level of service by reminding the service } people that when it's time to replace our scopes, service history will have } a MAJOR bearing on whose machines we buy. However, I'm extremely reluctant } to turn a previously very pleasant relationship into a confrontational, } grudging one (although it seems to be headed that way, anyway). In } addition, I can understand why some of these things are happening. If I put } myself in the shoes of the OEM service managers, I'm not sure what I would } do differently. On the other hand, I also have a tiny, nagging feeling that } part of these problems are deliberately designed to pressure us back into } our old contracts. That possibility (and I don't know it to be true) annoys } me intensely, but I still haven't played the sales managers against service } managers card. My sense is that OEMs make big bucks on service contracts } and they take it VERY personally when we "fire them" as one service manager } phrased it. } } I would like to emphasize very strongly that when field service engineers } come out, they are uniformly excellent. They do everything they are allowed } to do to provide top-notch care for the instruments. The problems } originate higher up. } } In summary, we have found that service management companies do indeed save } money on comparable contracts, but we have had a miserable time getting } service at all when using them. This may not be typical---I don't know. } Our level of service has declined greatly in terms of promptness, attitude, } and parts supply. The attitude part was certainly not worth the extra } several thousand dollars the OEMs wanted, but the promptness and parts } issues probably were. Most disturbing was finding out that we could be } denied service because of a problem originating at another university in } another state that had absolutely nothing to do with us. } } We will continue for now with our new IC's to see if the problems correct } themselves, especially since the problems didn't arise with this company. } In addition, one of the OEM service managers called and suggested we might } try a custom contract in which we could work with them to design our own } service schedule. He said we could be informally trained during service } visits to perform many of the maintenance chores on the scope and could } retain a contract that would basically cover emergencies, at a substantial } cost savings. We are looking into this now, because it seems to have lots } of nice possibilities. (We already do filament/gun exchanges and cleanings, } obviously, but I've never personally done a mechanical column alignment or } routine maintenance requiring major disassembly.) } } My best advice: if you are currently with an OEM, stay there. If you are } losing your local in-house service wizard and need to decide on contracts, } go with the OEMs. Fight like hell to resist pressure to switch. } } If you are with an insurance company and are happy with them, stay there, } and I wish you continued good luck. Please tell me your secret. } } My suggestion to OEM service providers: offer some in-house maintenance } training courses, better maintenance manuals and schematics, and encourage } customers to do more maintenance tasks (with appropriate provisos for } covering dumb mistakes). Lower your contract prices so we're not so } pressured to drop them. The glory days are over, folks. Time to compete. } } As for me, I have no financial interest in any of these folks. I just want } our scopes to work. Feel free to contact me with any questions. } } } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } }
Hayat cites Brack, C.(1973), Experientia, 29:76 ("Use of uranyl formate [0.5%] staining for the electron microscopic visualization of DNA-protein complexes").
You might also check, Bremer, et al.(1994), J Mol Bio, 742:683-700, and Steinmetz et al., M(1998), JMB,276:1-6.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy email: fmonson-at-wcupa.edu Phone: 610-738-0437
} ---------- } From: Chen Chen } Sent: Thursday, August 23, 2001 3:14 PM } To: microscopy-at-sparc5.microscopy.com } Subject: uranyl formate } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, everyone, } Does anyone have the recipe and protocol on how to make uranyl formate } solution for negative staining? } And do you have any idea about whhich is better, uranyl aceate or formate? } Thanks a lot. } } Chen Chen } } Dept of Biol. Chem. } the Johns Hopkins Univ. } School of Medicine } } }
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At 01:13 PM 8/23/2001 -0500, Macatangay, Peggy J., Celanese/US wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a PGT EDS / imaging system on and ISI DS130. The PGT sales people are very nice, the applications folks helpful, and their service man is highly competent - but I truly regret buying their system. The short version of the story is that this particular system (from 1997) uses a PC front end to run the Sparc board that really runs the EDS and imaging software, and the PC and Sparc *hate* each other. This is a cumbersome, frustrating construct; maybe later versions are less unsatisfactory.
If anyone wants to know more please let me know off-list, as I am uncomfortable being so negative in a public forum. Unless that is bad netiquette - maybe it is unprincipled to take it to private e-mail, and ought to be said in plain view. Whatever...
Disclaimer: These comments are my personal observations and opinions and do not reflect or represent the views (if any) of my employer.
Regards, Andrew T. Werner Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
As was pointed out to me by another person, I neglected to discuss third-party service providers. This is mainly because I have no experience with them, but they are increasingly becoming an attractive alternative. The bulk of my posting, I think, argued for retention of OEM contracts whenever possible, for many of the reasons you discussed below. In our experience, the difference in service between insurance and OEMs is incredible.
That said, however, it is still a mystery to me why OEMs seem to consider billable work at $200 or more per hour, including travel time, plus per diem, mileage, hotels, meals, etc. to be some kind of sacrifice that deserves punishment. To me that seems like a pretty good bonus over what they are likely to make under a service contract. One recent visit to service our FESEM resulted in charges of well over $7000, mostly to work with software glitches! Two more visits like that and we will have paid for the OEM contract and the OEM will have increased their income over what a contract would have brought them. Maybe it's simply that money up front is preferable, as you say, since that eliminates the risk of not being paid at all.
If working on a billable basis really is a hardship for OEMs and we're told we can't afford their service contracts, then that's where the free market comes in and third-party engineers have found their niche. People like you fill that gap admirably. I expect life isn't always easy, though, especially when you must rely on OEMs for parts and specialized expertise. One third-party provider told me of losing thousands of dollars when an OEM changed the price of a part upon finding out that it was ordered by an independent service provider. I've also heard that sometimes OEMs try to avoid selling parts to independents. Have you had any experiences along these lines?
It seems that the entire service landscape is changing and everyone is scrambling for alternatives. Makes for interesting discussion.
Randy
-----Original Message----- } From: Ken Converse [mailto:qualityimages-at-netrax.net] Sent: Thursday, August 23, 2001 9:53 PM To: Tindall, Randy D.; MSA, listserver
As much in any area, it is important to just get your hands on a unit and try it out and see if it will do what you want in the way you want. Most systems will have the capabilities you want, but sometimes there are restrictions or encumbrances that can build up frustration over time.
I believe there is also an issue of corporate heritage and influence. As a rule, larger, more established companies _should_ have and devote the resources necessary for new developments. They should have good, polished products which continue to improve. There should also be good support behind those products over many years. Smaller, newer companies are generally innovative and should be expected to come up with new product ideas, but the execution of those ideas might be a little rough. They do not have the resources to polish everything nicely. There might also be issues of support over the long run. There is also a good chance that a smaller company might be swallowed up by a larger one.
This seems to be proving true in the EDS field, but there are exceptions to some of these principles with almost every company. Some you would expect to deliver more but they don't, and vice versa. We currently have an Oxford ISIS and an IXRF EDS system. We also had a Kevex Delta (became part of Noran) and a Tracor TN-2000 (became Noran) systems. We looked at EDAX and PGT and they had good systems, but we made other choices. That has been a few years ago. I would have to investigate the field thoroughly if I were making the decision again.
I don't think that we regret any of the decisions that we made under the circumstances of the time. We have been satisfied with the systems we chose. I suppose you could say that in various ways, the companies did not keep up or made some strategic choices of their own and we opted for different systems when it came time for replacement. (Some of that is now ancient history. For example, Tracor used a proprietary operating environment on its 5000 series of analyzers and that was a major negative for us. They have since gone more standard.) Sometimes it was an issue of price-performance ratio for the features we were getting.
So I come back to my earlier position. -Find out the capabilities of the current systems. (Qual, Quant, Imaging, Mapping, Linescans, etc.) -Determine which of those are really important to you and necessary. -Evaluate your options for those functions for accuracy AND usability and price. Be sure to get your own hands on the units during the demos. It make take a couple looks at the models to give a fair look. -Then make your choice.
Happy hunting. Warren
At 01:13 PM 8/23/2001 -0500, you wrote:
} I am investigating EDS systems to integrate with a JEOL-6100 SEM. I'm } leaning towards PGT's Spirit system. Other considerations are Oxford's INCA } and Thermo Noran's Vantage. Does anyone have particularly positive or } negative experience with PGT? I don't often hear much about that company. } Any comments about the other possibilities would be appreciated, as well. } } Thanks! } } -Peggy } } Peggy McKarns Macatangay, PhD } Project Analyst 2 } Celanese Corpus Christi Technical Center
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Regarding this subject, the local Zeiss rep gave me a copy of an interesting and valuable article that hits right on the topic. The article is "Choosing Objective Lenses: The importance of numerical aperture and magnification in digital optical microscopy." Piston, D. (August, 1998). The Biological Bulletin, 195/1.
Anyone interested should look this up. It also talks about digital capture and the relationship to laser scanning confocal microscopes.
Sorry I am so long in posting this but a strange and wonderful thing intervened - summer vacation.
Below are three of the responses I received on the question of HMDS. We used the following protocol and the sample seems to show good morphology. The gross morphological features I think are probably affected by mechanical forces when you remove "flimsy" structures from any liquid. The ciliated structures appear well preserved.
Every step of the protocol is done in a fume hood especially for the HMDS, which is a strong irritant.
A. After the fixation (in 1G4F), the dehydration begins with two baths of one hour for each step. 1. PO4 wash 2. 50% ethanol 3. 70% ethanol 4. 80% ethanol 5. 95% ethanol 6. 100% ethanol
C. Pure HMDS 1. HMDS 1 - 1 hour 2. HMDS 2 - 1 hour
D. The samples are placed fresh HMDS in a dessicator with silica gel in the bottom overnight. They are ready to be dissected.
Celine Barre is the researcher looking at the specimens.
"O'Neil, David" wrote: } } We have been critical point drying scallop specimens successfully for SEM to } examine them as they grow. We are now at a point where the specimens no } longer fit in the CPD chamber. With only a couple of specimens to go we } thought to try HMDS. The samples are approximately 30mm in diameter. Does } someone have a protocol they have used for larger specimens that has worked } for them? You can send them directly to me and I can summarize for the } list. Thanks in advance.
Hi David, I have not worked on scallops but I do regularly work on fleshy invertebrates - polychetes, snails, insects etc. My method is to take the specimen through OH dehydration series 50, 70, 80, 90, 100, 100%. At each stage I leave the specimen in for a few hours. Because the specimen is large in diameter and quite thick (5mm). Then 50% HMDS + 50%OH for several hours, I do change the solution 2 to 3 times during this period. Then finally into fresh 100% HMDS for several hours. To dry I remove the specimen from the HMDS and place it in a petri dish that has filter paper on the bottom of it which is lightly soaked with HMDS. The lid sits over the top - this ensures that the drying is slow and controlled. If the drying is too rapid I find that some specimens distort. Oh all of this procedure must be done in a fume hood. Another little bit is that when soaking in 50% and 100% HMDS be careful when removing the lid. There is often a gas buildup in the bottle. I think the gas is NH4???? Not very pleasant. Also, you might find that the scallop is too thick for successful penetration from both HMDS and OH. If this happens then you might have to consider halving the specimen so to reduce the thickness thus allowing better penetration. I hope this helps???
Have fun
Sue
-- Sue Lindsay
SEM Laboratory Manager Scanning Electron Microscope Unit The Australian Museum ph 02 9320 6198 6 College st fax 02 9320 6059 Sydney, NSW, Australia Email suelind-at-austmus.gov.au
Hi David I don't know what CPD you have available, but I've dried crocodile pharyngeal specimens (ca. 5X8X1.5cm) quite successfully in a BioRad CPD Jumbo. I remove the basket holder from the CPD chamber and fill up with Eth-OH while tilting the apparatus, door uppermost and then transfer the specimens into the chamber and seal it. Specimens are "manipulated" onto the floor by tilting the whole bomb back and fore. Drying is done over an extended period of about 3.5 days. Naturally a large volume of CO2 is used flushing every ±2-3h through the day. I have a circulating waterbath and do the run at 10oC.
I'm interested to see how other folk overcome large specimen drying.
Regards John
Mr John F. Putterill Electron Microscopy Unit Tel: (Int) 27-12-529-9175 Pathology Section Fax: (Int) 27-12-529-9165 Onderstepoort Veterinary Institute E-mail: john-at-moon.ovi.ac.za Private Bag X05 http://www.ovi.ac.za Onderstepoort 0110 South Africa
I've been watching the listserver for several days for replies to your inquiry, and haven't seen any. I've been using HMDS since the 1980's, and have found that it works well within certain constraints. It is not something to use if you need to preserve the size of a sample, as it causes shrinkage of tissues, particularly of connective tissues, that is not seen to a great extent with critical point drying. HMDS is caustic, and flammable, and should be used only in a fume hood by someone with gloves on. It produces samples with good surface morphology, similar to CPD. With samples such as you are talking about, you will need extensive time to infiltrate your scallops in order to replace your dehydrating fluid (ethanol) with the HMDS. Otherwise, you will find that the ethanol will still be in the center of your sample and will not give you good drying results. If I were doing your project, I would try to slice the scallops in cross section into several pieces so the fluids you use can get in and out of the tissue more easily, or cut tissue out of the bottom of the scallops (since this side will be mounted on your sample holder and not used for imaging). Make sure to dispose of the waste HMDS in a sealed container so the highly irritating fumes don't get out in your lab.
For tissue like rabbit ovaries, I bisect the ovaries longitudinally after glutaraldehyde fixation. These ovaries are 1.5cm long, but only 0.5cm in diameter. It is not necessary to use osmium tetroxide for HMDS preps. I then rinse my tissue for 15 minutes in buffer, then 15 minutes in 0.9 % salline, the idea here being to slowly change the osmotic pressure seen by the tissue. I use phosphate buffer, and so must then go through 3 10 minute rinses in distilled water to insure complete removal of phosphate salts from the tissue. If this step is skipped , the salts will precipitate on the outside of the tissue when drying in HMDS, a phenomenon I didn't see when doing CPD.
Following the water rinse, I dehydrate the tissue in changes of ethanol, 15-30 minutes per change, in 35%, 70%, 95%, and two half hour changes in 100%. I infiltrate my samples with 100% HMDS, 2 times, for a minimum of 10 minutes each time. Following infiltration, I remove the HMDS from my sample vial and then shake my samples out on a paper towel or filter paper so they will dry, moving them several times so all sides dry quickly. The tissue will turn white colored as it dries, a process that can take up to a minute for larger samples. I work with all of my solutions at room temperature, and do all of my work in the hood. For dense samples such as the ovaries, or your scallops, once the sample appears visibly dry I transfer it to a 60 Degree vacuum oven for several hours or overnight to insure that the sample is completely dry before sputter caoting it for the SEM. If you don't have access to a vacuum oven, a vacuum is still better than letting the sample remain at atmospheric pressure to dry.
HMDS is said to harden the tissue while it is infiltrating it after ethanol dehydration. I don't know if the chemical interaction of HMDS and acetone or methanol will produce the same hardening effect. This is why I only dehydrate with ethanol.
I envy people like you that get to work with edibles. I met some people in Florida that study reproduction in Florida lobsters, and others that study the same thing in grouper. These poor folks have to "discard" the carcasses of their study subjects after removing their gonads. Tough life, but somebody has to do it! I only get to work with human and lab animal tissue. Oh well! Let me know if this helps you. Feel free to write with other questions if you need to.
Edward Haller Lab Manager Diagnostic Electron Microscopy University of South Florida Pathology Department (813)974-9584
hi matthew, There is a monoclonal antibody commercially available that recognizes callose (1-3)-B-glucans), if you want to do immunogold labeling. It's easy to use - it doesn't care if the tissue is embedded in Epon (Embed 812) or if it has been fix with osmium. The company is Biosupplies Australia. Visit their web site for more info www.biosupplies.com.au The product is Cat # 400-2. it's a good one so you should have good luck with your project, Beth
} Subject: Ask-A-Microscopist:stain for b eta glucans mainly callose
} Email: mcs4-at-dana.ucc.nau.edu } Name: Matthew Salanga } } Organization: Northern Arizona University } } Education: Graduate College } } Location: Flagstaff, Arizona } } Question: I have been doing TEM work on plant cells for my thesis. I } was wondering if anyone might have a suggestion on a stain which } would be specific for } beta glucans mainly callose. Literature suggests that normal UA and } lead citrate do not stain callose at all, however it appears as } though it may according to what I believe is Callose in my } micrographs. Anyways if anyone has suggestions or comments please } email me. } } Thanks!
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
I think any EDS system search would be somewhat incomplete without taking a look at and evaluating some of the smaller companies that provide PC based systems coupled with new EDS detectors. In the final analysis, it seems to come down to a system that best fits your application and needs, ease of usability and performance. Last but certainly not least is future support.
Some of these companies that come to mind would be WinEDS by TNAS, 4pi Analysis, and Emispec Systems. There are, of course, others that provide these systems. Sometimes dealing with smaller companies can be an advantage.
Good Luck,
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 480.967.3946
Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } As much in any area, it is important to just get your hands on a unit } and try it out and see if it will do what you want in the way you want. } Most systems will have the capabilities you want, but sometimes there } are restrictions or encumbrances that can build up frustration over time. } } I believe there is also an issue of corporate heritage and influence. As } a rule, larger, more established companies _should_ have and devote the } resources necessary for new developments. They should have good, } polished products which continue to improve. There should also be good } support behind those products over many years. Smaller, newer companies } are generally innovative and should be expected to come up with new } product ideas, but the execution of those ideas might be a little rough. } They do not have the resources to polish everything nicely. There might } also be issues of support over the long run. There is also a good chance } that a smaller company might be swallowed up by a larger one. } } This seems to be proving true in the EDS field, but there are exceptions } to some of these principles with almost every company. Some you would } expect to deliver more but they don't, and vice versa. We currently have } an Oxford ISIS and an IXRF EDS system. We also had a Kevex Delta (became } part of Noran) and a Tracor TN-2000 (became Noran) systems. We looked at } EDAX and PGT and they had good systems, but we made other choices. That } has been a few years ago. I would have to investigate the field } thoroughly if I were making the decision again. } } I don't think that we regret any of the decisions that we made under the } circumstances of the time. We have been satisfied with the systems we } chose. I suppose you could say that in various ways, the companies did } not keep up or made some strategic choices of their own and we opted for } different systems when it came time for replacement. (Some of that is } now ancient history. For example, Tracor used a proprietary operating } environment on its 5000 series of analyzers and that was a major } negative for us. They have since gone more standard.) Sometimes it was } an issue of price-performance ratio for the features we were getting. } } So I come back to my earlier position. } -Find out the capabilities of the current systems. (Qual, Quant, } Imaging, Mapping, Linescans, etc.) } -Determine which of those are really important to you and necessary. } -Evaluate your options for those functions for accuracy AND usability } and price. Be sure to get your own hands on the units during the demos. } It make take a couple looks at the models to give a fair look. } -Then make your choice. } } Happy hunting. } Warren } } At 01:13 PM 8/23/2001 -0500, you wrote: } } } I am investigating EDS systems to integrate with a JEOL-6100 SEM. I'm } } leaning towards PGT's Spirit system. Other considerations are } } Oxford's INCA } } and Thermo Noran's Vantage. Does anyone have particularly positive or } } negative experience with PGT? I don't often hear much about that } } company. } } Any comments about the other possibilities would be appreciated, as well. } } } } Thanks! } } } } -Peggy } } } } Peggy McKarns Macatangay, PhD } } Project Analyst 2 } } Celanese Corpus Christi Technical Center } } } ---------------------- } Warren E. Straszheim } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } }
Mike, The technique is "energy-dispersive spectroscopy" (in contrast to wavelength-dispersive spectroscopy). In formal writing, there is little reason to abbreviate these terms, but of course folks do. It is possible that EDX describes a proprietry instrument that some OEM named in this way.
Hope this helps, Tobias (no problem is too small to baffle me) Baskin
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Either one is correct. It's a matter of taste. EDXS would be more correct (Energy Dispersive X-ray Spectroscopy). Williams&Carter suggest yet another one - XEDS. I use EDX.
Max Sidorov AMD
-----Original Message----- } From: Ingram, Mike [mailto:MIngram-at-rodel.com] Sent: Friday, August 24, 2001 11:58 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Not a serious question here, more of a curiosity.
I use EDS when talking about x-ray analysis. I see many using EDX. Which is correct?
I have always used EDS because of the counterpoint to WDS. But I always define its first use as Energy Dispersive X-ray Spectroscopy. Now, I am going to start using Nestor's convention and start referring to both of them as X-ray Energy Dispersive Spectroscopy (XEDS) and X-ray Wavelength Dispersive Spectroscopy (XWDS). Hey I have an idea --let's start a convention!
BTW, it should never be EDAX. That's the same as calling all photocopies, Xeroxes.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ingram, Mike [mailto:MIngram-at-rodel.com] Sent: Friday, August 24, 2001 2:58 PM To: 'Microscopy-at-MSA.Microscopy.Com'
Not a serious question here, more of a curiosity.
I use EDS when talking about x-ray analysis. I see many using EDX. Which is correct?
I have used what I believe is the generic version of XEDS, x-ray energy dispersive spectroscopy.
At 2:58 PM -0400 8/24/2001, Ingram, Mike wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
When I was shopping for one, I was corrected by someone along the way that the "S" in EDS stands for spectrometry, not spectroscopy. Several books here in the lab library also use that term.
Before I confuse part of the next generation of microscopists this upcoming semester, is one term better than the other - or is it a potato, potatoh, tomato, tomahtah (let's call the whole thing off) kind of thing?
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
The best material we've found to use is a polyester mounting media such as our Polymet material. In fact, I just did exactly that for some office "fun" around here. We found a nice little black widow that ended up making an even nicer paperweight. I will send you information off-line on that product.
Best regards-
David
john_bruss-at-bose.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (john_bruss-at-bose.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } August 22, 2001 at 17:00:52 } --------------------------------------------------------------------------- } } Email: john_bruss-at-bose.com } Name: John Bruss } } Organization: Bose Corp. } } Education: Graduate College } } Location: San Diego, CA, USA } } Question: What materials and process would an amateur use to suspend } a spider in a clear cube for unmagnified artistic and historic use? } Where are those materials available in small quantities? } } ---------------------------------------------------------------------------
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
This name problem has been around for 30 years! Back in 1971 John Russ edited a little book called Energy Dispersive Analysis of X-rays published by the ASTM. Then Nuclear Diodes, Inc, renamed themselves EDAX and the competition could not use that trademarked name. John worked for EDAX and promoted the technique as "EDAX". Kevex had a book published in 1973 called "Everything you wanted to know about XES (X-Ray Energy Spectrometry)" but that name did not stick. EDS and EDX have stuck and are the most popular. I once wrote a paper on filter use for "EDXRF" (Energy Dispersive X-Ray Fluorescence) and it did not get listed in abstract indexes of X-Ray papers because they did not recognize the name! I think EDS is a bad name, but I use it because others understand what I mean.
Mike, My guess is that either will do the trick, but I would take EDX as a shortened version of Energy Dispersive Analysis of X-rays, which, of course, is EDAX. I've also seen EDXA which would keep you out of trouble with EDAX. One really is doing a specific type of analysis, spectroscopy, which would cause me to lean towards Energy Dispersive Spectroscopy.
Ken Converse owner Quality Images third party SEM service Delta, PA
Ingram, Mike wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Not a serious question here, more of a curiosity. } } I use EDS when talking about x-ray analysis. I see many using EDX. Which } is correct? } } Mike Ingram } } }
While this is off topic I am getting enough email from on the subject I think has enough interest to the group that I should post the particulars on sources for the CD-ROM. My only interest in the project is I suggested the idea to Shawn Carson and I am very glad that they carried it out. It has all the Amateur Scientist articles from the 20's until very recently.
I got mine from CD-ROM http://www.surplusshed.com/ the Surplus shack for $39 dollars. Fry's Electronics is a west coast electronics chain http://www.frys.com/ they don't sell from their web but the Sunny Vale store does sell mail order their phone number is (408) 617-1300. Fry's has them for $25 dollars and I have been told that they have a mail in $25 dollar rebate that comes with it.
There are still a few places left for he New York Microscopical Society Workshop listed below: N.Y.M.S. Bernard Freidman Memorial Workshop Use of the Microscope, September 15,22,29,Oct. 6,2001,10AM to 4PM
A basic course on light microscopy which will cover the following topics: T heory of microscopy, Kohler Illumination, Diffraction Theory, Contrast Methods , Polarized light, Phase Contrast, Interference, Hoffman contrast, Rheinberg, Dark-field & oblique Illumination, etc. The workshop will consist of four consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of microscopy. The course instructors include Jan Hinsch of Leica, Inc., Dennis O’Leary of Micro-Optical Methods, Mary McCann of McCann Imaging, John Reffner of SensIR Technologies, Inc. and N.Y.M.S. Instructor Don O'Leary.
WHERE: 30 3. Mountain Avenue, Montclair, NJ, Phone (973) 744-0043 ( parking, accessible by public transportation, Information on car pools and transportation will be provided.) COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.HOW: Register using the form below. Limited to the first 12 registrants.Send form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410. FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849 E-mail donoleary-at-worldnet.att.net Fax (425) 988-1415
PLEASE POST ---------------------------------------------------------------------------- ------------------------------------------------------------------ Registration Form, Use of the Microscope
When I was shopping for one, I was corrected by someone along the way that the "S" in EDS stands for spectrometry, not spectroscopy. Several books here in the lab library also use that term.
Before I confuse part of the next generation of microscopists this upcoming semester, is one term better than the other - or is it a potato, potatoh, tomato, tomahtah (let's call the whole thing off) kind of thing?
Dear Heather, The "scopy" part has to do with observation, whereas the "metry" part has to do with measurement (i.e., quantitation), so energy-dispersive spectroscopy is separating the photons by energy in order to look at the spectrum, and E-D spectrometry is separating the photons in order to perform measurements. I can understand a vendor insisting that the instrument is capable of quantitation and is, thus, a spectrometer. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
We no longer need our Quantimet 970 Image Analysis System interfaced with an intensified Vidicon – type camera. The camera was bottom-mounted on a TEM. We are the original owners, have kept it well serviced (contract), and have used this system up until the last year. Interested parties please contact tcgruber1-at-excite.com offline.
Regards,
Tyler Gruber
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Does anyone know if gridded coverslips are available made out of Thermonox?
Thanks,
David Spector -- Dr. David L. Spector Cold Spring Harbor Laboratory One Bungtown Road Cold Spring Harbor, New York 11724 Tel. (516) 367-8456 Fax (516) 367-8876 email: spector-at-cshl.org
A couple of years ago, our company was contacted by lawyers from "Electronic Data Systems" (EDS) who told us that we were violating their trademark by offering a product called "The Personal EDS". I personally thought this was nonsense, but it was going to cost us a lot of money to pursue this, even if we were able to prevail.
So I now use EDX because: (1) it is recognized; and (2) I don't much like talking to lawyers.
If it were up to me, I would much prefer "XES" (which as Ron Vane pointed out, was the name Kevex used to use). "X-ray Energy Spectroscopy" may not be completely self-explanatory, but at least it is not misleading. The use of the word "dispersive" in this context is just plain wrong! (you are not "dispersing" energy) EDS does have a "symmetry" with WDS, but it is an inappropriate one IMHO.
Fred Schamber ASPEX, LLC
"Ingram, Mike" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Not a serious question here, more of a curiosity. } } I use EDS when talking about x-ray analysis. I see many using EDX. Which } is correct? } } Mike Ingram
An excellent Philips CM-10 transmission electron microscope is being offered for sale by Michigan State University. It is 1987 vintage and has been on service contract since new. Asking price: $10,000. Please contact Dr. Xudong Fan for more detail, fanx-at-msu.edu, 517-353-4525.
Yours Sincerely, Xudong Fan, Ph.D. Center for Advanced Microscopy Michigan State University East Lansing, MI, 48824
Looking for a replacement RGB monitor for my LINK eXL system. Oxford has been contacted and due to the "very high" price to get a replacement "for this rare jewel" I am exploring other options. I know the power supply is damaged because of all the burnt components but am not sure what else may be damaged. Fix may take a while, so I'm hopeful someone may have a retired system with a monitor I could deal for. Interest from third party EDS repair companies who would like to tackle this beast would also be useful.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Chen Chen: Uranyl formate, in my experience using it as a negative stain to reveal the finest detail of virus structures at very high mags, gave superior results to uranyl acetate. In short, there appeared to be a finer grain detail and a better resolution of the virus's structure. A word of caution: uranyl formate should be used within about 15 minutes after making it as it tends to readily form a precipitate. For general work where I'm not trying to obtain the ultimate detail, I stick to uranyl acetate simply to avoid the precipitate problem. My protocol is to make up a 1% sol. in dd water and immediately give a brief spin in a centrifuge. Some have simply filtered it with a Whatman filter. Draw off just the top portion for staining. I find a final solution of 0.5% works ok. I place a drop of my virus solution on parafilm, and after a few minuets wait, touch my grid to the surface to pick up the phage. I find that the phage tends to collect on the surface of the drop. After removing most of the phage containing drop I apply a drop of stain or, if you wish to "cleanse" your sample prior to staining, you may apply a drop of a 1 mM solution of ammonium acetate in dd water for a very brief period. Depending on the sample, I've sometimes follow up with another brief application of a drop of dd water. The uranyl formate negative stain is applied and all but a trace immediately removed. Keep in mind that beam exposure does effect the stain and delicate virus structure. I try to use low dose exposure. Choose something off to the side of your sample to focus on, and if necessary, do a through-focus series to avoid creating the appearance of a substructure artifact. If your uranyl formate or uranyl acetate is in a bottle that has sat on the self for donkey's years, look to see that it is of uniform color and not peppered by greenish flecks. If it is not of a uniform color, you may find it has difficulty going into solution, indicating it time to order a new bottle. Hope this helps, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences State University of New York at Binghamton Binghamton, NY 13902-6000
A colleague is looking for an electron micrograph (SEM and/or TEM) of a cockroach cercus (or cerci) to show students in our undergraduate physiology laboratory, who are carrying out electrophysiological measurements with this structure. If you have a picture we could use, we would be very grateful, and would, naturally, give you full credit.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
I'll add my thanks for your viewpoint. The problem we in Universities are having is this: you wrote } "University service } labs are not supposed to be profit centers. They are } SERVICE labs and } are heavily subsidized...."
Sadly, survey says that very few of us have administrators who share your viewpoint. You make excellent points about all of the options, and we can only hope this entire discussion helps when renewal time comes around. Perhaps the OEM Service departments need to "suit up", bring a fancy presentation, and state their case against the ICs.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Thursday, August 23, 2001 10:53 PM, Ken Converse [SMTP:qualityimages-at-netrax.net] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Randy, } Thank you. This is quite interesting. } } I'd like to respond from the viewpoint of one who worked } for an OEM for } 4 years and has subsequently had his own third party } service company for } 20 years. } } First, the OEM is obligated to service contract customers } first. You } have become a "billable" call and DO go to the bottom of } the list. That } would not be all that different with a third party firm } (although they } might try a little harder to be timely). The contract } people have paid } up front (or have at least committed up front to pay) for } an entire } year. This is guaranteed income and includes a guaranteed } liability } (for the OEM) "to maintain the instrument/system to its } original } specifications". Service organizations have to take this } seriously. } This liability doesn't exist for billable customers. } Work will be done } to the best of the OEM's ability and not beyond the limit } of the } purchase order. } } Second, the IC has no technical expertise, no intimate } connection with } its customers and no obligation to use the manufacturer. } I have been } called by at least one IC several times, but have never } done any work } for them because they haven't a clue what I'm talking } about when I tell } them I service SEMs but not TEMs and I like to stay within } 500 miles of } home. (Their geography hasn't impressed me, either). } They are trying } to emulate what happens when your car gets in a fender } bender. There } are lots of people who can fix it, and many of them can do } a pretty } satisfactory job. But ask yourself, "When my car has a } rumble, or the } engine misfires and I need expert help or maintenance, who } do I call? } My insurance company?" Probably not, not only because } your insurance } doesn't cover that kind of thing, but because they } couldn't help you } anyways. They know about insurance, not the inner } workings of your car. } } Third, any outside party that starts making commitments } FOR the OEM (you } won't have to pay for recertification..............) I } would turn away } from and run, not walk, run. If they tell you that THEY } will pay for } recertification if you're unhappy and they will put it in } writing, } that's another story. } } As to the manufacturers making lots of money on their } contracts, some } may, some may not. A lot depends on how well they are set } up and run. } It also depends on the instrument density. If one } engineer can service } 20 or 30 systems within driving distance, the engineer is } competent and } the service department backs him up well, yes, it can be } quite } profitable. If you have to fly to every site, you're } short on } experienced engineers and you don't support them well, you } can lose your } shirt ( and have a whole lot of PO'd customers to boot). } } Every time I've gotten one of those glib calls from an IC } in Texas, in } the back of my mind a little voice is always saying "And } just what makes } you think you're going to get paid?" I do thank you for } confirming that } this concern is warranted. } } The question now comes down to "What do you want?". The } OEM has reason } to feel miffed. You want their expertise, their parts, } their top level } response, their best guess on advance parts replacement } for reliability } and you're going to pay someone ELSE the bonus if they do } all this. The } IC is betting that the instrument will run reliably as it } always has, } meaning very little cash out. The OEM no longer has the } incentive to go } out of its way to insure reliability. They'll actually } make more if } the system breaks more frequently. And as I pointed out, } if they do } their best work, you pay someone else the bonus while they } still have } the considerable expense of running a service department. } I'm not } implying in any way that they will go out of their way to } make it fail. } One of the advantages of sticking with the organization } that actually } does the servicing is that you establish a raport with the } personnel in } the field and in house. I'm only saying that they aren't } going to go } out of their way to KEEP it from failing, because it's not } in their } interest to do that. It IS in their interest when they } have guaranteed } the performance for a full year. } } Yes, generally speaking my service contracts are } profitable, but there } is another major reason that I prefer to have systems on } contract. It } becomes up to me how much time I spend on an instrument } and how fast or } slow I work on it. Sometimes I'm very sure what the } problem is and fix } it immediately, but often there are intermittent problems } and I can tell } you that a billable customer watches the clock like a hawk } when he sees } me sitting in front of a running system waiting for an } intermittent } problem to show up. Sometimes there are subtle problems } that the } customer isn't even aware of, yet. If the system is on } contract I can } work without watching the clock and get everything right. } In the long } run it saves me repeat trips. Also, if I should decide to } let something } slide due to whatever circumstances, I know that I'm } guaranteeing the } work and a later failure will be fixed at MY expense, not } the customer's. } } There was a time when people seemed more likely to reward } good service } by paying a premium for it and showing a little loyalty to } those who } provided it. The loyalty seems to be going by the board, } and more and } more people want more for less, they aren't willing to pay } for quality } because in their everyday life they buy cheap and when it } breaks they } throw it out and buy cheap again. I don't believe that } the scientific } instrument market will get there any time soon. This is } specialty } equipment, capital investment, there for the long haul.. } If it's well } taken care of , it will last for decades. That's where } you can get the } savings. } } If you were happy with the quality of the OEM's service } and they are } willing to work with you, by all means see what you can } work out! If } you weren't so happy with an OEM, see if there is a third } party service } company that you can establish a good relationship with. } The ICs can't } charge less, take out a middleman's cut, and give the } organizations who } actually do the work and support the equipment, enough } money to maintain } the quality you demand. If it looks too good to be } true................................................... } } I sympathize with the cost-cutting that is being forced on } you from the } bean-counters, but beware of false economies. Maybe one } or more } instruments could be taken off contract, but what kind of } grants are you } (the school and faculty) going to lose if it's down too } much of the } time? What are the priorities of the organization? } University service } labs are not supposed to be profit centers. They are } SERVICE labs and } are heavily subsidized because the typical student (grad } or undergrad) } can't afford to pay commercial rates for instrument time } (especially } with what they're paying for tuition). If these } instruments are } important to various departments then the decision is too } important to } be made by some MBA who doesn't know the difference } between hydrogen } embrittlement and cilia. } } It would be interesting if we could get a response from } the insurance } industry side of things. } } My $.02 worth. } } Sincerely, } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } } } Tindall, Randy D. wrote: } } } } } ---------------------------------------------------- ----- } } --------------- } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } ml } } } } --------------------------------------------------------- } } --------------. } } } } } } As promised, here is a rundown on our OEM vs. insurance } } experiences. I } } decided to put this on the list based on the very large } } number of replies I } } received, some asking specifically that this be posted. } } As a result, I've } } removed all names, since I don't have a clue as to legal } } ramifications of } } being more specific. Sorry to be so wimpy, but in these } } cases I believe in } } CYA. } } } } The following obviously reflects our experiences alone, } } but based upon what } } I have heard from a few others, I don't believe that our } } situation is } } unique, by any means. } } } } Our lab, like many others, has been under increasing } } pressure to cut costs } } for the last two to three years, and since service } } contracts represent a } } significant proportion of our budget, they logically } } became a cost-cutting } } target immediately. It was beginning to appear that we } } were going to lose } } the contract on one of our TEM's to save money, } } especially since it wasn't } } heavily used. About this time one insurance company } } (IC, for short) began a } } series of high-powered presentations to the University } } purchasing people and } } administrators, promising significant cost reduction, } } with no reduction in } } services, choice of service providers, elimination of } } paperwork, etc. } } Facilities like ours were contacted by University } } purchasing staff in order } } to set up individual meetings to assess our needs and } } get estimates on the } } IC's costs versus OEM charges. } } } } After attending two open seminars and having two } } meetings with the insurance } } people and purchasing staff in our facility, the } } decision was made, with my } } agreement, to give the IC a shot at managing our } } service. The thinking was } } that we would try it for a year. The IC said we could } } back out at any time } } with no penalty, they would pro-rate our contracts to } } adjust for the } } differing ending dates for our OEM contracts, and, they } } said, there should } } be no "re-certification fee" if we decided to go back to } } OEM contracts, } } since we intended to use only the OEMs' engineers } } anyway. They would take } } care of the paperwork notifying the OEMs about the new } } arrangement, and all } } billing would be handled between the IC and the service } } providers. They } } would guarantee no increase in contract costs for, I } } believe, three years. } } } } The very first service call we made to the manufacturer } } on one of our TEMs } } resulted in our being told that they couldn't send an } } engineer, because we } } weren't on service contract any longer. When I } } explained that they were } } supposed to have been notified that we were with the IC, } } I was told that we } } would have to provide a University P.O. before anyone } } would be sent. } } Apparently, there was a substantial outstanding bill } } from this IC for } } service provided to a university in another state, and } } until it was paid no } } service would be provided on our contract. The IC's on- } } campus rep, our } } purchasing director, and the director of the university } } core facilities were } } notified by me immediately. Apparently some high level } } phone calling went } } on, because before the day was out, the service provider } } had called back } } saying the outstanding bill would be paid and an } } engineer was on the way. } } (Additionally, our purchasing director assured us that } } we could get a P.O. } } if necessary. We have terrific support here from } } purchasing and they have } } been invaluable on several occasions.) } } } } The engineer was sent out, the scope was back on line. } } BUT, it turns out } } that the promised payment was never made and we were } } back to square one the } } next time we needed service on that machine. In fact, } } it was worse, because } } the OEM was never paid for the service on our scope } } either. } } } } Our other scopes are from a different manufacturer and } } initially service was } } pretty much the same as before (although they made it } } clear they weren't } } happy about the switch). Response time was a bit } } slower, and there was more } } stinginess with replacement parts, but we had no real } } complaints. } } } } Then, the IC declared bankruptcy. Their campus reps } } disappeared, their web } } site went down, and they effectively vanished. We had } } already used part of } } the year's contract, but were only able to recover about } } $700 on the unused } } portion. Some labs lost thousands, I've been told. The } } service provider } } was never paid, and lost a substantial amount of money. } } (I'm told that this } } IC has reorganized under a different name and is } } soliciting new business, } } especially from universities. Ask lots of questions if } } a new company comes } } around. Be especially careful of large numbers of } } enthusiastic people in } } nice suits giving slick multi-media presentations!) } } } } We began discussing returning to our old OEM contracts } } and I was asked to } } get prices. The first thing I was told by one service } } provider was that we } } would have to have our instrument re-certified, which } } would cost about } } $1500, even though nobody had touched that scope but the } } OEM engineers. } } Needless to say, that rankled a bit, so we checked into } } another insurance } } company that had been recommended by several other labs. } } We were } } immediately impressed by the fact that they presented } } what seemed to be a } } more realistic picture of what they could do for us. } } They also offered } } significant savings (10-15%) over the old contracts, so } } we once again } } decided to give it a try. } } } } This time, the first call we made had the same result as } } before, i.e., the } } provider wanted a P.O. before an engineer would come } } out. When I asked why, } } they said that even though they had no problems with the } } new IC, they } } weren't going to take a chance on losing a ton of money } } again. If the } } university would sign a waiver accepting responsibility } } for paying any } } amounts in default, we could get service again. Fine, } } I said. Several } } days later the form was faxed to us and I forwarded it } } to our purchasing } } director, who responded that he had passed it along for } } review to the people } } who needed to sign off on it. That was last month } } sometime, and I have } } heard nothing yet. So far we have been waiting to get a } } preventive } } maintenance visit on this scope for about two months and } } nothing's in sight, } } yet. } } } } On our other scopes, we have again been able to get } } service, but now it } } seems that we are way, way down on the priority list. } } OEMs will tell you } } that they are obligated to service their contract } } customers first, and in } } our experience they certainly do. The service } } management (insurance) } } companies will tell you that this is not the case, that } } it's first-come, } } first-served. That has not been our experience. On one } } TEM we have been } } waiting weeks for a PM. It was scheduled twice and an } } engineer was here, } } but was pulled away on "emergency" calls, which I read } } as calls from } } contract holders. The PM has not yet been done. (Just } } minutes ago we } } received a call from our service engineer saying he's } } been pre-empted again } } and now he has NO idea when he can get here.) Our SEM } } has been serviced } } under the new arrangement in a relatively timely manner. } } } } } } Under the OEM contracts, service was provided } } "yesterday" and parts were } } replaced if they were even suspected to be bad. The } } working relationship } } was wonderful, and the service was stellar. We were } } called and reminded of } } PMs before they were due and we received periodic phone } } calls just to check } } on the status of our equipment. Under the new } } contracts, service is } } provided (if we can get it) whenever they get around to } } it, and engineers } } can be interrupted by calls to respond to contract } } holders. Parts are } } changed less frequently and are all billed. We schedule } } the PMs (no big } } deal). The relationship is much more tense than it used } } to be. } } } } We could probably increase the level of service by } } reminding the service } } people that when it's time to replace our scopes, } } service history will have } } a MAJOR bearing on whose machines we buy. However, I'm } } extremely reluctant } } to turn a previously very pleasant relationship into a } } confrontational, } } grudging one (although it seems to be headed that way, } } anyway). In } } addition, I can understand why some of these things are } } happening. If I put } } myself in the shoes of the OEM service managers, I'm not } } sure what I would } } do differently. On the other hand, I also have a tiny, } } nagging feeling that } } part of these problems are deliberately designed to } } pressure us back into } } our old contracts. That possibility (and I don't know } } it to be true) annoys } } me intensely, but I still haven't played the sales } } managers against service } } managers card. My sense is that OEMs make big bucks on } } service contracts } } and they take it VERY personally when we "fire them" as } } one service manager } } phrased it. } } } } I would like to emphasize very strongly that when field } } service engineers } } come out, they are uniformly excellent. They do } } everything they are allowed } } to do to provide top-notch care for the instruments. } } The problems } } originate higher up. } } } } In summary, we have found that service management } } companies do indeed save } } money on comparable contracts, but we have had a } } miserable time getting } } service at all when using them. This may not be } } typical---I don't know. } } Our level of service has declined greatly in terms of } } promptness, attitude, } } and parts supply. The attitude part was certainly not } } worth the extra } } several thousand dollars the OEMs wanted, but the } } promptness and parts } } issues probably were. Most disturbing was finding out } } that we could be } } denied service because of a problem originating at } } another university in } } another state that had absolutely nothing to do with us. } } } } } } We will continue for now with our new IC's to see if the } } problems correct } } themselves, especially since the problems didn't arise } } with this company. } } In addition, one of the OEM service managers called and } } suggested we might } } try a custom contract in which we could work with them } } to design our own } } service schedule. He said we could be informally } } trained during service } } visits to perform many of the maintenance chores on the } } scope and could } } retain a contract that would basically cover } } emergencies, at a substantial } } cost savings. We are looking into this now, because it } } seems to have lots } } of nice possibilities. (We already do filament/gun } } exchanges and cleanings, } } obviously, but I've never personally done a mechanical } } column alignment or } } routine maintenance requiring major disassembly.) } } } } My best advice: if you are currently with an OEM, stay } } there. If you are } } losing your local in-house service wizard and need to } } decide on contracts, } } go with the OEMs. Fight like hell to resist pressure to } } switch. } } } } If you are with an insurance company and are happy with } } them, stay there, } } and I wish you continued good luck. Please tell me your } } secret. } } } } My suggestion to OEM service providers: offer some in- } } house maintenance } } training courses, better maintenance manuals and } } schematics, and encourage } } customers to do more maintenance tasks (with appropriate } } provisos for } } covering dumb mistakes). Lower your contract prices so } } we're not so } } pressured to drop them. The glory days are over, folks. } } Time to compete. } } } } As for me, I have no financial interest in any of these } } folks. I just want } } our scopes to work. Feel free to contact me with any } } questions. } } } } } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } } } } } } } } }
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Dear Listers, I need your suggestions about the possiblity to buy a system based on the inverted microscope, but suitable for deconvolution and time-lapse analysis of fast events in living cells.
We would like to buy such system. Whinch one is the best (I mean quality devided on price)?
Sincerely yours, Alexander Mironov Consorzio Mario Negri Sud, Italy
At 05:26 PM 8/28/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We (Nanoprobes) sell methylamine vanadate as a 2 % aqueous solution (suitable for use directly) - the product is called "NanoVan." It's listed on our web site catalog under "Negative staining."
Regards,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
Try: Joseph S. Wall, Ph.D.
516-344-2912; Fax: 516-344-3407
E-mail: wall-at-stem.bio.bnl.gov Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Jeannette Taylor } Sent: Tuesday, August 28, 2001 6:26 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: query methyl amine vanadate } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would appreciate information on methyl amine vanadate and where to buy } it. It is used as a negative stain. } } Thank you, } Jeannette Taylor } }
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Jeannette Taylor } Sent: Tuesday, August 28, 2001 6:26 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: query methyl amine vanadate } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would appreciate information on methyl amine vanadate and where to buy } it. It is used as a negative stain. } } Thank you, } Jeannette Taylor } }
A post-doctoral position is immediately available to work on the development of new computational methods in image processing. The candidate will join a multidisciplinary team working in the field of structural biology. Our goal is to determine the three-dimensional structures of macromolecular complexes using high-resolution electron cryo-microscopy. Electron microscopy is a rapidly developing technique to explore a large number of cellular structures with medical and biological importance. Our laboratory has a well-established electron microscope facility, and equipment for protein purification, sample preparation, and image processing. Image data are currently being generated by several projects in our laboratory. Processing is performed on our UNIX alpha and SGI workstations using software partly developed in our laboratory. The role of the post-doctoral researcher will be to improve existing, and develop new image processing algorithms to push protein structure determination by electron microscopy towards atomic resolution. One important objective will be to develop a detailed understanding of electron scattering from biological samples, and to develop algorithms to extract information from electron scattering patterns and images.
The ideal candidate would have a background in applied physics or material sciences. Candidates from other fields with experience in microscopy, as well as light, electron or X-ray scattering techniques are also encouraged to apply.
Formal applications, which should include a CV, statement of research interests, and the names, addresses and e-mails of 3 references, should be sent to
Dr. Nikolaus Grigorieff Brandeis University - MS029 415 South Street Waltham, MA02454-9110 USA
I hope you will indulge me with a moment of your time.
I'm involved with a group here in Wichita that's working on some water treatment technologies that reduce chemical usage as compared to current
treatment methodologies. We also are working on a system using a version of these same new technologies to reduce algae growth in pond water. I've been conscripted to help them assemble the "water lab" for experimentation and testing and generally manage the development project.
Well, I've done plenty of development projects, but my knowledge of microscopy is pretty much limited to the Skillcraft 'scope I had as a child and the low magnification bench scopes I've set up for microelectronics manufacturing. I am absolutely delighted to have found
your microscopy.com group and have been "cramming" for the past week or so. I think I'm now ready to ask you a couple of questions. (and maybe make sense...)
The two main (and quite different) thrusts of microscopic analysis that we're attacking first are determining the polymorphic state that calcium
carbonate is present in, in potable water (aragonite or calcite) and the
viability of algae that has been exposed to "treated" water in a pond setting. What I'm most curious about is your (expert) opinion as to the best strategies to image these two areas of interest, and to do so out to a high- quality digital imaging system. (Nikon, SPOT RT, etc.)
After nucleation, the calcium carbonate in question will either be in colloidal suspension in liquid water or will be dried onto a slide. I'm betting that a water immersion objective would be your recommendation for the liquid viewing, but I'd rather hear it from you. Sure, if we had an electron microscope, we could image individual seed crystals, BUT a) we don't have that kind of budget and b) we're pretty sure we can induce nucleation and crystal growth on demand, so we'll just grow 'em until we can see 'em. I guess what I'm more curious about is whether you feel that good old fashioned transmissive light microscopy is the right path or whether we'd see significant advantage in exploring polarizing light, phase contrast, DIC
or some other kinky form of imaging to best see the orthorhombic nature of the calcite or the hexagonal prismatic aragonite. For what it's worth, I've read somewhere of using a special setup to actually derive the refractive index of the sample in question. Aragonite does have a different value in this respect than does calcite - in fact aragonite's value is higher than the two values for calcite (birefringent?). Eeek! I now know just enough to hurt myself and not much more...
As for the algae, we're trying to show that we have a process for killing it, so primarily we're looking at ways to image healthy (pre-treatment) specimens and then treat the water and show (over time) that we really do kill the stuff. Again, same question - is there some creative way to do this with good 'ol planapochromatics or do I need a pricey Fluorescence setup to make the chlorophyll twinkle? OR "What exactly does freshly dead or dying algae look like and how do you best look at it?".
We'll be doing some particle sizing and Zeta potential analysis as well,
so we're not trying to be cheap on the microscopy side - BUT, I don't want to buy more or less than we need and the 'scope salesmen all think we need everything they sell...
Any light you have time to shed on these issues will be greatly appreciated - if there's a chance of speaking with you on these issues or on the possibility that either you or someone you respect being available for some consulting work in these areas, please let me know and I'll gladly set up a conference call. Much gratitude for whatever you can do!
Best regards,
Jeff -- Jeff Hedlesky Aladdin Companies Wichita, KS
Hi All, I'm hoping that someone out there thinks that in situ localization using non-radioactive DIG-labelled RNA probes on sections of tissue on glass slides is almost as easy as flipping a light switch. I would greatly appreciate some advice as I've been banging my head against the wall for months trying to get this to work. The background is that I'm doing the in situ work with plant tissue, namely roots that have been lightly fixed in a paraformaldehyde/glut. mixture (as per in situ chapter in Steven Ruzin's bible of plant microtechnique) and embedded in butyl-methyl-methacrylate as per Tobias Baskin's methods (thanks again Tobias). After some struggle with the molecular biology side of the experiment, I found that the gene I was given to work from had been subcloned along with a bit of "junk" DNA from the old plasmid it had been in. I used PCR to amplify ~100 bp pieces of that gene to get around the "junk" and to avoid having to hydrolyse my probes, since I often lose a large amount of probe when I hydrolyse. The questions I have relate more to the detection side of the matter. I've been using about 4ng of RNA probe per slide (as recommended in the Ruzin bible), then using an anti-DIG (Fab fragments) antibody conjugated to alkaline phosphatase supplied by Roche. Then I "develop" the labelling using the standard NBT-BCIP detection used often-times for Western blots. The controls work pretty well at this point. My main question is, has anyone used fluorescently conjugated anti-DIG antibodies for this type of work (or a secondary fluorescent conjugate)? It seems that it would be much more straight-forward than guessing when the NBT-BCIP reaction has reached a good level of staining. Any other insights/advice would be appreciated. Thanks again for your help, Kristen
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011
I am trying to section with a cryo diamond knife that has a boat. I am not sure on what solution to use in the boat. So far we have tried a DMSO and ETOH, 50/50 mixture. This slowly froze over. Please help:) Melissa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kai-at-lehigh.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 29, 2001 at 15:34:48 ---------------------------------------------------------------------------
Email: kai-at-lehigh.edu Name: Kai Lorcharoensery
Organization: Lehigh University
Education: Graduate College
Location: Bethlehem, PA, USA
Question: TEM sample preparation: I would like to see the structure of Fe particle cross-section. These particles are about 40-100 um in size. I tried casting them with epoxy in 3-mm copper tube but either particles fell off or epoxy thin film (100 um) was detached from Cu ring.
The preparation steps are 1. Casting 2. Slicing the Cu tube with high speed diamond saw 3. Grinding with 600 grit- 8 um- 3 um SiC paper 4. Polishing with 1 um Al2O3 on nylon cloth ==} ~ 80-100 um and here is where the epoxy disk was usually detached from the Cu ring.
Position Announcement Chemical and Materials Engineering Dept. Arizona State University
A post-doctoral candidate is sought to work on an interdisciplinary research project to study the interaction of functional molecular monolayers with nanostructured arrays on electronic materials. A new interface force microscope will be constructed and studies conducted on molecular monolayers on quantum dots, wires, etc. Emphasis will be placed on biomimitic molecules containing functional and transductional components with potential for electronic applications. Position available immediately. Please send cover letter and resume including the names, addresses and phone or e-mail of 3 professional references to picraux-at-asu.edu {mailto:picraux-at-asu.edu} . Preference will be given to applications received by September 14, 2001 and will be continued until the position is filled.
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John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Perhaps with controlled nucleation and experience you could induce recognizable and differentiable calcite and aragonite forms to grow. However, these materials can take on a wide variety of crystal habits depending upon the other ionic species and concentrations present, temperature and organic materials present. Often acicular spherical crystal groups will form and the individual crystals will not be distinguishable with sufficient clarity to determine their optical properties. It is also possible to arrive at situations where mixed phase agglomerates form.
Calcium carbonate is subject to the influence of organic materials which strongly influence crystal morphology by preferentially binding to one or another of the crystal faces. In this way the aspect ratio of the crystals may change dramatically from one batch to another if trace organics vary. Other organics may influence the degree of saturation of the solution with respect to calcium carbonate. For example, most sea water is undersaturated with respect to calcium carbonate even though vast amounts of limestone occur in contact with it. One reason is that a very small amount of free fatty acid is sufficient to complex with the exterior of the carbonate phase, in effect isolating the solid surface and interrupting the solid {==} solution equilibrium.
To get an idea of how varied the crystal habit of this material can be (and to gain an appreciation for living in the age of X-ray diffraction!) you might take a look at the classic work on morphological description by Victor Goldschmidt, "Atlas der Krystallformen". There is an entire volume out of the library-shelf-size "Atlas" on calcite.
Good luck.
John Twilley Conservation Scientist
Jeff Hedlesky wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopy Wisemen (and women!): } } I hope you will indulge me with a moment of your time. } } I'm involved with a group here in Wichita that's working on some water } treatment technologies that reduce chemical usage as compared to current } } treatment methodologies. We also are working on a system using a } version of these same new technologies to reduce algae growth in pond } water. I've been conscripted to help them assemble the "water lab" for } experimentation and testing and generally manage the development } project. } } Well, I've done plenty of development projects, but my knowledge of } microscopy is pretty much limited to the Skillcraft 'scope I had as a } child and the low magnification bench scopes I've set up for } microelectronics manufacturing. I am absolutely delighted to have found } } your microscopy.com group and have been "cramming" for the past } week or so. I think I'm now ready to ask you a couple of questions. } (and maybe make sense...) } } The two main (and quite different) thrusts of microscopic analysis that } we're attacking first are determining the polymorphic state that calcium } } carbonate is present in, in potable water (aragonite or calcite) and the } } viability of algae that has been exposed to "treated" water in a pond } setting. } What I'm most curious about is your (expert) opinion as to the best } strategies to image these two areas of interest, and to do so out to a } high- } quality digital imaging system. (Nikon, SPOT RT, etc.) } } After nucleation, the calcium carbonate in question will either be in } colloidal suspension in liquid water or will be dried onto a slide. I'm } betting } that a water immersion objective would be your recommendation for the } liquid viewing, but I'd rather hear it from you. Sure, if we had an } electron } microscope, we could image individual seed crystals, BUT a) we don't } have } that kind of budget and b) we're pretty sure we can induce nucleation } and } crystal growth on demand, so we'll just grow 'em until we can see 'em. } I } guess what I'm more curious about is whether you feel that good old } fashioned transmissive light microscopy is the right path or whether } we'd see } significant advantage in exploring polarizing light, phase contrast, DIC } } or some other kinky form of imaging to best see the orthorhombic nature } of the calcite or the hexagonal prismatic aragonite. For what it's } worth, I've read somewhere of using a special setup to actually derive } the } refractive index of the sample in question. Aragonite does have a } different } value in this respect than does calcite - in fact aragonite's value is } higher } than the two values for calcite (birefringent?). Eeek! I now know } just enough to hurt myself and not much more... } } As for the algae, we're trying to show that we have a process for } killing it, so primarily we're looking at ways to image healthy } (pre-treatment) specimens and then treat the water and show (over time) } that we really do kill the stuff. Again, same question - is there some } creative way to do this with good 'ol planapochromatics or do I need a } pricey Fluorescence setup to make the chlorophyll twinkle? OR "What } exactly does freshly dead or dying algae look like and how do you best } look at it?". } } We'll be doing some particle sizing and Zeta potential analysis as well, } } so we're not trying to be cheap on the microscopy side - BUT, I don't } want to buy more or less than we need and the 'scope salesmen all think } we need everything they sell... } } Any light you have time to shed on these issues will be greatly } appreciated - if there's a chance of speaking with you on these issues } or on the possibility that either you or someone you respect being } available for some consulting work in these areas, please let me know } and I'll } gladly set up a conference call. Much gratitude for whatever you can } do! } } Best regards, } } Jeff } -- } Jeff Hedlesky } Aladdin Companies } Wichita, KS } } } } }
Let me inform you that my so called "Petr's Microscopy Resources" have been moved to the new address
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The items Societies and Meetings are rebuilt. The other items (Literature, Database, Laboratories, Production & Sales, Courses & Software, Information Resources) have been modified, and will be transferred to the new technology too.
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I don't know anything about algae but there are other ways to solve your CaCO3 question. You are not interested in the shapes or sizes of the particles, right ? So IR or Raman spectroscopy might be the way to go, these are two references on that topic: Calcium carbonate phase analysis using XRD and FT-Raman spectroscopy. Kontoyannis, Christos G.; Vagenas, Nikos V. Inst. Chem. Eng. High Temperature Chem. Processes, Dep. Pharm., FORTH, University of Patras, Patras, Greece. Analyst (Cambridge, U. K.) (2000), 125(2), 251-255. Simpson, L. J. Electrochemically generated CaCO3 deposits on iron studied with FTIR and Raman spectroscopy. Electrochim. Acta (1998), 43(16-17), 2543-2547. There are other references, but that should do for a start. There is also a fancy tool called IR or Raman microscope. This thing combines optical microscopy and optical spectrometer. Therefore you can take Raman or IR spectra from individual particles and see which ones are vaterite or aragonite or whatever phase you are looking for. I've done that with ZnO particles about 500 nm to 1 micron in size, so I actually know it works.
Hope this helps Andreas
************************************************* Dr. Andreas Taubert Dept. of Materials Science and Engineering 3231 Walnut Street University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
----- Original Message ----- } From: Jeff Hedlesky {jhedlesky-at-edgebb.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 29, 2001 1:12 PM
Hi Listers...
Does anyone know when the first cold field emission microscope became commercially available?
Thanks and best,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
1) A "coupling agent" to promote adhesion of the copper to the epoxy. I have used Dow Z-640 (originally meant as a coupling agent for fibers in fiber glass, I believe), to reduce pullout of particulates during microtomy. This was originally suggested by Klinger and Schwab (I can look up the full reference if you are interested).
2) Incorporating the particulate into a metal foil instead of an epoxy. This involves either electroless nickel plating or electroplating of a sufficiently thick film that it can then be handled as a "bulk" sample. Either give good adhesion to the particles (at least in the case of the aluminum powders or ceramic particles with which I am familiar). Electroless plating has the benefit of bonding to non conductive materials. You might look at Morra et al (Materials Sci. and Eng. A124, 1990, "A Technique for prep. of powders...") as a reference.
kai-at-lehigh.ed u () To: Microscopy-at-sparc5.microscopy.com cc: 08/29/01 Subject: Ask-A-Microscopist: TEM sample preparation 06:57 PM
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kai-at-lehigh.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 29, 2001 at 15:34:48 ---------------------------------------------------------------------------
Email: kai-at-lehigh.edu Name: Kai Lorcharoensery
Organization: Lehigh University
Education: Graduate College
Location: Bethlehem, PA, USA
Question: TEM sample preparation: I would like to see the structure of Fe particle cross-section. These particles are about 40-100 um in size. I tried casting them with epoxy in 3-mm copper tube but either particles fell off or epoxy thin film (100 um) was detached from Cu ring.
The preparation steps are 1. Casting 2. Slicing the Cu tube with high speed diamond saw 3. Grinding with 600 grit- 8 um- 3 um SiC paper 4. Polishing with 1 um Al2O3 on nylon cloth ==} ~ 80-100 um and here is where the epoxy disk was usually detached from the Cu ring.
********************************************* This Email contains Important Information about the Microscopy Listserver. Please read it all then SAVE a copy for future reference. - Nestor ****************************************
The message you all received was a blunder.. too many hours on-line and my fingers and mouse start wandering aimlessly missing the correct keys. It was supposed to go out to today's set of Newsubscribers only, not the entire list.
} Melissa: It depends on what temperature you are using when } sectioning. When I am sectioning a temperatures that freeze the DMSO mix } I use ethanol. Ethanol will stay a liquid down to about -95C. With our } microtome you can control temp of knife and boat and gas separately so I } can keep the knife temp to about -90C and can still have the sample a } little lower than -120C. This enables me to section siloxanes. If you } need to go colder than this I think you will have to try and section } dry. Hope this helps. Steve } } I am trying to section with a cryo diamond knife that has a boat. I } am not sure on what solution to use in the boat. So far we have tried a } DMSO and ETOH, 50/50 mixture. This slowly froze over. Please help:) } Melissa
Stephen McCartney Research Associate 2108 Hahn Hall Materials Institute Virginia Tech Blacksburg, VA 24061-0344 USA
I am looking at an advertisement from Coats and Welter Instruments for "The world's first commercial field emission scanning electron microscope." It is touted as ...operating at room temperature and not being subject to burnout as are the conventional hot filament electron emitters... While the ad is not dated it does reference a paper from 1968 so it was probably produced sometime in the late 60's or very early 70's. Greg
Angela Klaus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Listers... } } Does anyone know when the first cold field emission microscope became } commercially available? } } Thanks and best, } } Angela } } Angela V. Klaus } } Director, Interdepartmental Laboratories } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024 USA } } Email: avklaus-at-amnh.org } Tel: 212-769-5977
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
I think we would ALL love to see tuition supporting inter-departmental facilities. Departments are not profit centers per se, but may be expected to be self-sufficient. And from what (little) I'm learning about administrative thought processes...your childrens' tuition is essentially "credited" to the departments (or school) in which they major. Maybe not directly as hard dollars, but in terms of "weight" to be thrown around :) Inter-departmental, multi-user facility, interdisciplinary....great buzzwords but no real money.
My $0.02...based on broad impressions of OTHERS' experiences. (I don't yet know whether I am describing the views of my employer....)
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami mlynn-at-miami.edu
On Wednesday, August 29, 2001 9:09 PM, Ken Converse [SMTP:qualityimages-at-netrax.net] wrote: } Matt, } I'll repeat what I said to Phil Oshel: } If I found a school turning any profit on a service lab, } I'd scream } bloody murder as I've got 2 in college and 2 on the way. } I would expect } my tuition bills to go a long way towards general support. } } } My take as a parent. } } Ken Converse } owner (and parent of 4) } Quality Images } third party SEM service } Delta, PA } } } Matthew Lynn wrote: } } } } } --------------------------------------------------------- } } --------------- } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } ml } } } } --------------------------------------------------------- } } --------------. } } } } } } Ken, } } } } I'll add my thanks for your viewpoint. The problem we } } in } } Universities are having is this: you wrote } } } } } "University service } } } labs are not supposed to be profit centers. They are } } } SERVICE labs and } } } are heavily subsidized...." } } } } } } Sadly, survey says that very few of us have } } administrators } } who share your viewpoint. You make excellent points } } about } } all of the options, and we can only hope this entire } } discussion helps when renewal time comes around. } } Perhaps } } the OEM Service departments need to "suit up", bring a } } fancy presentation, and state their case against the } } ICs. } } } } Matt } } } } Matthew J. Lynn } } Center for Advanced Microscopy } } University of Miami } } (305)284-4736 } } mlynn-at-miami.edu } } } }
Since our JSM 25 died, we are looking for a good used SEM preferably in the following models:
1. Hitachi S570
2. JOEL no older than 10 years
3. Philips no older than 10 years
Please contact me at: tyoho-at-lhup.edu
--------------------------------------------------------- Tim P. Yoho, Professor Yoho (Joho) Amateur Radio: WA3D Department of Biology First Known Use in 1395 (570) 893-2391 Lock Haven University Origin: Switzerland & Alsace Fax: (570) 893-2047 Lock Haven, PA. 17745 http://www.lhup.edu/~tyoho tyoho-at-lhup.edu ---------------------------------------------------------
Dear Kai, When doing samples like you have described, I usually grind to 100 microns using a Disc Grinder from Gatan, or the same device from other specimen prep companies, on 1200 grit paper stuck on a sheet of glass. If detachment is your problem, then cast your sample/epoxy into a slightly larger copper tube with INSIDE diameter of 3 mm. and glue a large-mesh grid or slot grid onto the exposed end of your sample when it is still fairly thick. Then detachment is desirable and will leave your sample stuck onto a grid for final thinning, dimple polishing, ion thinning or whatever you wish to attain transparency. At 05:57 PM 8/29/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
An international conference and exhibition on the science of microscopy and in-situ analysis; a unique opportunity for you to hear about topics at the cutting edge of microscopy and to see for yourself the very latest developments in light, scanning probe and electron microscopes, associated equipment and image analysis systems in both life and physical sciences.
The Science...
The conference programme will be a series of Invited Lectures by internationally acclaimed experts, which will be closely linked to Tutorials, Poster Sessions and special "hands-on" Workshops.
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fluoresence & molecular markers confocal & multiphoton microscopy FLIM & FRET low-temperature microscopy variable pressure SEM tomography digital image acquisition and analysis scanning probe microscopy energy filtered imaging EBSD FEGTEM SuperSTEM
The Exhibition.....
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For more information, contact the RMS by email
Info-at-rms.org.uk
Or visit the website
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-- _____________________________ Dr. Rik Brydson, Leeds Electron Microscopy and Spectroscopy (LEMAS) Centre Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
I'm having a problem with my MT-1B Ultra microtome. When cutting ultra thin sections my microtome skips cutting a section completely then cuts a section that's too thick. Any solutions? I can't find a company that services Sorvall MT-1B microtomes. Anyone have a source for service calls. Bill
Fourier transform infrared microspectroscopy (a.k.a. infrared microscopy) may prove more useful to your work than polarized light microscopy to differentiate and identify polymorphs of calcium carbonate. The former provides chemical analysis of molecular composition, while the latter provides examination of morphology and measurement of optical properties.
Infrared microscopy couples an optical microscope to an FTIR spectrometer. The optical microscope is used to position small areas of a specimen in a condensed infrared beam for analysis. The FTIR spectrometer gives an infrared spectrum that is used to identify molecular composition and to make qualitative and quantitative analyses of samples. The technique can distinguish between calcium carbonate polymorphs such as calcite and aragonite.
Most infrared microscopes are equipped for brightfield illumination and transmitted visible illumination. Most systems, like mine, also can be fitted for polarized light examination and brightfield fluorescence illumination, which can be useful in differentiating phases in specimens. Analyses can be made in transmission if the specimen is prepared on an infrared-transparent window material such as diamond, KBr, BaF2, etc., or by reflection-absorption if dispersed on a reflective surface such as a gold filter. Other methods are used, but may limit your ability to differentiate particles by morphology before analyses. Preliminary tests will help you develop a methodology.
If your specimen particles are individual and smaller than 5-10 micrometers in diameter, then you might want to investigate Raman microscopy, which complements infrared microscopy, but provides better spatial resolution (about 1 micrometer) and allows one to work through glass, at two or three times the cost ($150K to $250K. For this money, one might also consider an FTIR microscope equipped with a focal plane array detector.
Please feel free to contact me off-line if you have questions.
Jamie
James Martin Orion Analytical, LLC www.orionanalytical.com martin-at-orionanalytical.com 413-458-0233 fax 413-458-5542
----- Original Message ----- } From: "Jeff Hedlesky" {jhedlesky-at-edgebb.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 29, 2001 1:12 PM
Hi, Bill-
} I'm having a problem with my MT-1B Ultra microtome. When cutting ultra thin } sections my microtome skips cutting a section completely then cuts a section } that's too thick. Any solutions? I can't find a company that services } Sorvall MT-1B microtomes. Anyone have a source for service calls.
Where are you located? Nowhere near me, I'll bet, but there are still a few people out there who know their way around an MT-1.
Sometimes skipped sections means you are actually trying to section too thin, so you miss one and then the next one is thicker, then you miss again. Try sectioning thicker until you get every section, and see if they are of a reasonable thickness.
As for the microtome, open it up and take a look inside. It's relavtively simple (as compared to current models) and you may see something obvious. Check belts for wear and cracks, see if moving parts are dirty or chipped, etc. They are mechanical marvels unencumbered by electronics!
Good luck, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I agree with Greg--early 1970s. I purchased a Coates and Welter FE SEM in 1973. It was not the first FE model, but was the first scope to come off their line with the gun mounted on a cabinet separate from the controls (at my request). They subsequently went to that style for stability.
It operated "cold," i.e., at room temperature, and the electrons were extracted with the "field" below. If the tip became contaminated, the way to remove crud was to warm it up some (I don't remember the temperature, but it wasn't hot.) to burn away the contaminants.
It worked great from the perspective of length of filament life, no vacuum problems, high resolution, and low voltage capability, but getting service from the other coast when needed took forever. Frequently, I would describe a problem, and the engineer would ship me a board. We kept the SEM for about 10 years and sold it back to the company for parts. But we got some great shots with higher resolution (around 50 Angstroms routinely) than anybody else was getting at that time, and it was guaranteed at 100 Angstroms.
The company exchanged hands several times: first C & W: then Welter bought it out; then Coates bought it back. Then it became Nanometrics, and somebody else (?) bought the SEM technology. Maybe one of the present sales reps can help with the recent history--it might be LEO???
Hope this answers your question.
(I have no commercial interest in any of this.)
Sara
On Thu, 30 Aug 2001, Greg Strout wrote:
} Date: Thu, 30 Aug 2001 09:30:40 -0500 } From: Greg Strout {gstrout-at-ou.edu} } To: Angela Klaus {avklaus-at-amnh.org} } Cc: Microscopy List Server {Microscopy-at-sparc5.microscopy.com} } Subject: Re: FE-SEM: Historical Question } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am looking at an advertisement from Coats and Welter Instruments for "The } world's first commercial field emission scanning electron microscope." It } is touted as ...operating at room temperature and not being subject to } burnout as are the conventional hot filament electron emitters... } While the ad is not dated it does reference a paper from 1968 so it was } probably produced sometime in the late 60's or very early 70's. } Greg } } Angela Klaus wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hi Listers... } } } } Does anyone know when the first cold field emission microscope became } } commercially available? } } } } Thanks and best, } } } } Angela } } } } Angela V. Klaus } } } } Director, Interdepartmental Laboratories } } American Museum of Natural History } } Central Park West at 79th Street } } New York, NY 10024 USA } } } } Email: avklaus-at-amnh.org } } Tel: 212-769-5977 } } -- } ================================================================== } Greg Strout } Electron Microscopist, University of Oklahoma } WWW Virtual Library for Microscopy: } http://www.ou.edu/research/electron/www-vl/ } e-mail: gstrout-at-ou.edu } Opinions expressed herein are mine and not necessarily those of } the University of Oklahoma } ================================================================== } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
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Greetings, I would like to add a few words on the thread about spectroscopy vs spectrometry. Bill Tivol defined (see below) spectroscopy as producing spectra for visual inspection and spectrometry as producing spectra for measurment.
This is perfectly rational, nice and tidy, but like so many other things in language, does not seem consistent with usage. The OED defines spectroscopy as the study of spectra and does not have an entry for spectrometry. My big Websters (II ed, yes thanks) lists both, as does Collins, and both are defined as the science and study of spectra. And I would argue that there are really relatively few instances of a pure spectroscope, where all you do is look at the spectra, and equally few instances of a pure spectrometer (where you never see the spectra--think of all those peaks on the "EDS" computer screen). Basically, as spectroscopists (are there any spectrometrists??) we have the job of producing, interpreting, and measuring spectra, and spectroscopy has been used for years as a word to define that act. The word spectrometry has likewise been used, but I doubt whether the measuring-observing distinction can be reliably applied. If given a choice, I would use spectroscopy for the lot because it is the older word. But I suspect that various sub species have their own convention (thus it is always as far as I know FTIR microspectroscopy; and perhaps always energy dispersive spectrometry) and these fields will keep their conventions.
As ever, Tobias Baskin
} } When I was shopping for one, I was corrected by someone along the way that } the "S" in EDS stands for spectrometry, not spectroscopy. Several books } here in the lab library also use that term. } } Before I confuse part of the next generation of microscopists this } upcoming semester, is one term better than the other - or is it a potato, } potatoh, tomato, tomahtah (let's call the whole thing off) kind of thing? } } Dear Heather, } The "scopy" part has to do with observation, whereas the "metry" part } has to do with measurement (i.e., quantitation), so energy-dispersive } spectroscopy is separating the photons by energy in order to look at the } spectrum, and E-D spectrometry is separating the photons in order to } perform measurements. I can understand a vendor insisting that the } instrument is capable of quantitation and is, thus, a spectrometer. } Yours, } } Bill Tivol } Wadsworth Center } Albany NY } (518) 473-7399 WFT02-at-health.state.ny.us
In 1976 both, Siemens and Vacuum Generator marketed FESEMs. The Vacuum Generator unit was considerably cheaper and possibly better too. Incidentally, the Siemens unit became a fiasco and likely was a major reason for the company's board to abandon the building of electron microscopes a year or two later. Siemens had been the first commercial manufacturer of electron microscopes and built the best instruments in the 50th and early 60th. The Philips EM300 was the first serious challenge to Siemens. Michael Beer of Chicago published a great deal of FESEM developments in the early 70th. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, August 30, 2001 9:43 PM, Angela Klaus [SMTP:avklaus-at-amnh.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Listers... } } Does anyone know when the first cold field emission microscope became } commercially available? } } Thanks and best, } } Angela } } Angela V. Klaus } } Director, Interdepartmental Laboratories } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024 USA } } Email: avklaus-at-amnh.org } Tel: 212-769-5977 }
Nestor's little slip and his apology should make us realize how valuable Nestor and his service of the listserver is to us. I just want to say Thank You Nestor for all the hard work you do to keep us on-line and spam free. You help all of us to stay connected in a nice environment where we can ask our questions and communicate our ideas.
Thank You Nestor!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We are beginning preparations for the moving of our old Cambridge S200 SEM. Even though the poor gal is only moving about 500 yards, the transfer is to another building requiring removal of walls and the ever so delicate transfer by fork lift into the back of a truck. We need information on what is required and/or helpful in making sure the SEM successfully completes the trip. Thanks in advance.
David W. Troutt Corporate Analytical Services Voridian Company P.O. Box 1972 Kingsport, TN 37662-5150 Phone: 423-229-1993 Fax: 423-229-4558
Contact Microtome Service Company. Although they appear to only service MT2 &2B, they might have info or long lost spare parts for your beast. Good luck! john
MTS 7568 Florian Way Liverpool, New York 13088 315-451-1404
On 30 Aug 2001, at 13:40, Hanika, William wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I'm having a problem with my MT-1B Ultra microtome. When cutting } ultra thin sections my microtome skips cutting a section completely } then cuts a section that's too thick. Any solutions? I can't find a } company that services Sorvall MT-1B microtomes. Anyone have a source } for service calls. Bill
Sorry. The I left the subject line blank on the first letter blank. I thought most folks would delete it thinking it was an advertisement for a lifetime supply of Viagra.
We are beginning preparations for the moving of our old Cambridge S200 SEM. Even though the poor gal is only moving about 500 yards, the transfer is to another building requiring removal of walls and the ever so delicate transfer by fork lift into the back of a truck. We need information on what is required and/or helpful in making sure the SEM successfully completes the trip. Thanks in advance.
David W. Troutt Corporate Analytical Services Voridian Company P.O. Box 1972 Kingsport, TN 37662-5150 Phone: 423-229-1993 Fax: 423-229-4558
I would argue that there are really relatively few instances of a pure spectroscope, where all you do is look at the spectra, and equally few instances of a pure spectrometer (where you never see the spectra--think of all those peaks on the "EDS" computer screen). Dear Tobias, Perhaps WDS comes closer to a "pure spectrometer" than EDS, since one measures the counts for a given wavelength, then another, etc., but it is not necessary that an actual instrument be purely one or the other for the vendor or user to distinguish the major function by different words. In the much-much-lower energy region, the instrument mostly used today is a (wavelength-dispersive) spectrophotometer. Newton's original prism instrument was a spectroscope. (I suppose that avalanche photodiodes could be used to make an energy-dispersive spectrophotometer, but I have not heard of such a thing.)
This is perfectly rational, nice and tidy, but like so many other things in language, does not seem consistent with usage.
Of course, practical usage is always messier than language describing an ideal. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi Bill, What routine service have you administered, and how often and when last?
Fred Monson'
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Hanika, William } Sent: Thursday, August 30, 2001 2:40 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: MT 1B Maintenance } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm having a problem with my MT-1B Ultra microtome. When cutting ultra } thin } sections my microtome skips cutting a section completely then cuts a } section } that's too thick. Any solutions? I can't find a company that services } Sorvall MT-1B microtomes. Anyone have a source for service calls. } Bill } }
Bill (AND Tina!) just made me look inside MT-1, and I had a thought (what pain!). Who has one and still uses it?
If you do, just reply to this with the following information tacked on - if you want to. Let me know how you came to possess IT, and what you use it for that it does better than those that are newer (and BETTER?). Mine came from a cart on its way to the dumpster! (Imagine!), and I use it for really difficult stuff, though not that often. Usually it reclines in a display case so that no one else will mess with it.
After a while I'll return with a summary.
By the way, on the same cart was an MT2 with a Christensen Cryo System with 2 controllers. All still work!!! Thank you Kent! Also, they came with the books!
Fred
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I have a sample of dried porous silica that I would like to embed and thin section. Could anyone give me any advice on the best resin and protocol to use? The idea of using a methacrylate resin has been thrown about (the idea, not the resin - I can assure you), however I would be very grateful for any advice regarding any of the resins available. If this is a bit specific then reply off-line to the address below.
Thanks Pippa ---------------------- Pippa Hawes, EMU School of Chemistry University of Bristol Cantocks Close Bristol UK BS8 1TS Pippa.Hawes-at-bristol.ac.uk
I have an old bottle of Epoxy Solvent, bought from Polysciences under their catalogue number 0638. Does anyone know what this solvent contains? The company tells me that they cannot find this item in their records and probably have deleted any technical or MSDS information pertaining to it. I'd like to get rid of the approx. 300 ml of solvent left in the bottle but don't what, if any, hazards should be considered.
Thanks for any help you could offer.
Kevin Halcrow
Dr Kevin Halcrow, Telephone (506)-648-5567 Honorary Research Professor, Fax (506)-648-5811 Department of Biology, EMail Halcrow-at-unbsj.ca University of New Brunswick, P.O. Box 5050, Saint John, NB Canada, E2L 4L5
$0.000001 worth! My experience is that once a faculty member ascends to an administrative post his first instruction concerning his previous colleagues is that each is a cost center to be discounted as rapidly as possible to the lowest possible value. There is nothing that remains in the administrative jargon (Bill of Administrative Rights) that is not focused on gibberish and avoidance of responsibility. Since that philosophy reposes at the highest academic levels, it is no wonder what is being found somewhat lower down.
At academic institutions libraries are called resources, but they are treated as cost centers! If core labs have positive bottom lines, they are only ephemeral (with support from self-accounting measures). Such a situation can only occur when there is an influx of direct financial support to individuals linked (100%) to those facilities. If the grant winners are affiliated with departments, the departments split the gravy, and the core remains a cost center. Whatever support the core gets in large institutions is NOT direct. For the most part such support will ultimately derive from indirect monies. In this way the administrator can claim that the institution provides "direct" support ("Most of the institutional support derives from our commitment to excellence for all of our academic and research commitments," she said at the convocation.) leaving the core as a cost center - gaining no "credit" for its part in raising the money on which it lives. At how many places does the investigator pay and then repay for the use of research animal facilities. When indirect monies can be in the 70%'s of direct costs (usually salaries + benefits) on the grant budget, there is BIG money involved. For example, for $50,000 in salary and benefits to the researcher, 70% indirect costs would give $35,000 more to the institution to use almost as it pleases. This matter is purposefully complicated by the fact that each institution must negotiate with the government for its percentage and coverage. No matter what one says, however, personal experience will almost always not be universally applicable.
Indeed, that admission would suggest that there may still exist Deans who believe that their sole purpose is to serve the faculty at the pleasure of the President (and the President is second)! For the most part today, such individuals are not permitted to survive in administrative positions. For example, it was not the student who changed the University/College in the '60's, it was the administrator who saw a dwindling student population as a threat to the institution and translated the threat downward to the faculty member who was causing students to flunk out or keeping them from getting into medical school. Children immediately sensed this trend and happily blamed professors for their academic problems. Administrators began to depend on the course evaluations for evaluating faculty performance. Faculty members adapted, because they had no other recourse. Students sensed the trend and became even more militant. Faculty dropped standards, because they had no other choice. Students became unaware of the diminution of standards, having arrived with expectations and demands. Faculty were already bottomed out, and no one could do anything when teaching ended up "in the basement". If a student couldn't understand a math instructor from Eastern Europe, he/she knew that any complaint would be treated as discriminatory and thus, patiently ignored. If the instructor couldn't teach, students always had the option to take the same course with one of the other faculty in the department. Having two or more classes with 24 students each (the administrative holly grail) taking the same course with the same book never caused competition between instructors for the better course evaluation. (I've seen professors who cry about their love for the student and preen about their great evaluations promoted while their much more erudite colleagues are left behind. Then they become administrators!) Academic administrators. A very interesting group of folks.
The best example of adminstrator malfunction in our business is the "new" building mentality that came out of the new construction boom from the 60's to the '80's. For every new building there is an administrator who will turn off the cooling system in the Fall and the heating system in the Spring, especially when informed that such "air" conditioning systems are designed to have both functioning at once all of the time ("That's why we have sealed windows!"). There are even a few who have turned off the heat during Christmas (does anybody remember that mid-year break?) and let the plumbing freeze.
I once worked in a brand new building whose internal fire doors were tested a few weeks after occupancy. On attempting to re-open them, the new owners found that some of the ADMINISTRATIVE changes to the building design had removed the access doors. About half of the fire doors remain closed for the life of the building.
I worked at a large Eastern Academy for many years. During that time, the greatest pleasure was to be so distanced from academic administrators that I never knew any by face. Of course, one always managed to receive individualized or generic administrative threats but NEVER the administrative accolades (even for BIG grant awards!). Even with a large government grant, one is considered a cost center!
Big or small, the academic administrator will remain one thing for certain. He or she was one of us!
} ---------- } From: Matthew Lynn } Reply To: } Sent: Thursday, August 30, 2001 11:25 AM } To: 'Microscopy-at-MSA.Microscopy.com' } Subject: RE: OEM service vs. Insurance Companies---Long message } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ken, } } I think we would ALL love to see tuition supporting } inter-departmental facilities. Departments are not profit } centers per se, but may be expected to be self-sufficient. } And from what (little) I'm learning about administrative } thought processes...your childrens' tuition is essentially } "credited" to the departments (or school) in which they } major. Maybe not directly as hard dollars, but in terms of } "weight" to be thrown around :) Inter-departmental, } multi-user facility, interdisciplinary....great buzzwords } but no real money. } } My $0.02...based on broad impressions of OTHERS' } experiences. (I don't yet know whether I am describing the } views of my employer....) } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } mlynn-at-miami.edu } } } On Wednesday, August 29, 2001 9:09 PM, Ken Converse } [SMTP:qualityimages-at-netrax.net] wrote: } } Matt, } } I'll repeat what I said to Phil Oshel: } } If I found a school turning any profit on a service lab, } } I'd scream } } bloody murder as I've got 2 in college and 2 on the way. } } I would expect } } my tuition bills to go a long way towards general } support. } } } } } } My take as a parent. } } } } Ken Converse } } owner (and parent of 4) } } Quality Images } } third party SEM service } } Delta, PA } } } } } } Matthew Lynn wrote: } } } } } } } } } --------------------------------------------------------- } } } --------------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } } Society of America } } } To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } } } ml } } } } } } } --------------------------------------------------------- } } } --------------. } } } } } } } } } Ken, } } } } } } I'll add my thanks for your viewpoint. The problem we } } } in } } } Universities are having is this: you wrote } } } } } } } "University service } } } } labs are not supposed to be profit centers. They are } } } } SERVICE labs and } } } } are heavily subsidized...." } } } } } } } } } Sadly, survey says that very few of us have } } } administrators } } } who share your viewpoint. You make excellent points } } } about } } } all of the options, and we can only hope this entire } } } discussion helps when renewal time comes around. } } } Perhaps } } } the OEM Service departments need to "suit up", bring a } } } fancy presentation, and state their case against the } } } ICs. } } } } } } Matt } } } } } } Matthew J. Lynn } } } Center for Advanced Microscopy } } } University of Miami } } } (305)284-4736 } } } mlynn-at-miami.edu } } } } } } } } }
From root Fri Aug 31 11:12:21 2001 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA19523 for dist-Microscopy; Fri, 31 Aug 2001 10:50:19 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA19519 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 31 Aug 2001 10:49:46 -0500 (CDT) Received: from bingnet2.cc.binghamton.edu (bingnet2.cc.binghamton.edu [128.226.1.18]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA19508 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 31 Aug 2001 10:49:32 -0500 (CDT) Received: from eichelberger ([128.226.188.35]) by bingnet2.cc.binghamton.edu (8.11.4/8.11.4) with SMTP id f7VFkUE07108 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 31 Aug 2001 11:46:30 -0400 (EDT)
Dear Karen: After touching the grid to the drop containing viral particles, I draw off most of the fluid then add a drop of the 1 mM sol of ammonium acetate. If you find that you still end up with remains of dried buffer solution, try an additional change of ammonium acetate rinse. Never let the drop of any solution you add to the grid to dry until you reach the final drop of stain application. If you don't already use self-closing, anticapillary tweezers, I would recommend them for minimizing carry over between solution changes. The advantage of using PTA and ammonium molybdate negative stains is that their pH can be adjusted. PTA is known for giving good contrast. I use a 1 N KOH solution to adjust the pH. I use a final stain concentration of 1% PTA. Ammonium molybdate although it has less contrast, gives a very fine grain appearance which is good for small detail at high magnifications. I use a final concentration of 1% aqueous uranyl acetate negative stain which yields more contrast than ammonium molybdate. It also has the advantage of acting as a positive stain for chromatin material. Be aware that phosphate and cacodylate are precipitated by uranyl salts. So phage containing these buffers should undergo a number of droplet rinse changes to remove any traces of the buffer before staining. Good luck. Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences State University of New York-Binghamton Binghamton, NY 13902-6000
Hello, I've been going through setting up an Electroscan ESEM E3 in our laboratory and vacuum problems aside, we now have a stage that doesn't want to move. Anyone ever come across stage problems and have some suggestions that I may try for this instrument?
The stage will actually move if you give it a gentle push along a gear arm, but it often locks up in the Z direction, and sometimes X/Y. Auto calibrate allows the stage to move in X/Y, but it never calibrates the Z direction. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I would greatly appreciate if you could share your opinion about performance and reliability of the Bench Top Turbo III vacuum evaporator from Denton Vacuum, also alternative suggestions are more than welcome. We are looking currently to purchase a vacuum evaporator for general EM preparation capable of carbon and metal evaporation, glow discharge and thickness control. Cleanliness of the vacuum is our main concern and especially the fact that the Bench Top model does not have LN trap.
Thank you,
Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
To all who asked about this article, I have scanned it and saved it as an Adobe PDF file. It can be downloaded from http://gaugler.com/lmarticle_1.pdf using your browser.
If you have any problems, please let me know. In PDF, the whole article is about 1.33MB file size.
gary g.
At 02:21 AM 8/28/2001, you wrote:
} Dear Gary, } I'd be interested in this article, but do not have } access to the Biological Bulletin. } Do you have the articlein electronic form, or is ther } a URL that you can point me to? } } Regards, Jeremy Sanderson } jb_sanderson-at-yahoo.com } } } } Regarding this subject, the local Zeiss rep gave me } } a } } copy of an interesting and valuable article that } } hits } } right on the topic. The article is "Choosing } } Objective } } Lenses: The importance of numerical aperture } } and magnification in digital optical microscopy." } } Piston, D. (August, 1998). The Biological Bulletin, } } 195/1. } } } } Anyone interested should look this up. It also } } talks } } about digital capture and the relationship to laser } } scanning confocal microscopes. } } } gary g. } } } } } } ____________________________________________________________ } Do You Yahoo!? } Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk } or your free -at-yahoo.ie address at http://mail.yahoo.ie
Hello, I need to replace the SE scintillator on my SEM (JEOL 5900), and I was considering using a YAP scintillator instead of the typical phosphorous scintillator. Does anyone have any experience or suggestions on this? YAP is supposed to have a better S/N ratio, which is appealing to me. Most of my work is at 10kv on non-conducting oxides and at moderate magnification, however I'd really like improved performance at lower voltages, smaller spot sizes, and higher magnification, where the S/N ratio becomes a problem with my current system. Thank you for any input.
Brad Johnson Pacific Northwest National Lab P.O. Box 99, K6-24 Richland, WA 99352 voice: 509-372-1635 fax: 509-376-3108
Thanks are given to Nestor and many people whom I do know but help me a lot. I have been in this list for several years. There have been many great articles on electron microscoy, sample preparations and knowledges of instruments. I collected some of them, really valuable. I saw there were some summarizing articles on specific questions in the past. Do we have soemone who are classify technical articles and store somewere in an internet server? I saw questions raised on repeated subjects. If we have a server stored old, valuble articles for list members to access, that will be great. Recently, we decided to buy an electropolisher for our lab. We obtained several responses offline and they were really useful and presented clearly the advantages and disadvantages of each brand machine. I will write a summary. We believed that we made the right decision.
Thank you for your attention.
Good weekend,
Weiliang Gong Vitreous State Laboratory Catholic University of America Washington, D.C. 20064
I would be interested in hearing from anyone with experience transferring files from Tracor Northern (Noran) 5400 or 5500 systems. Many years ago I had written a FORTRAN program running on a DEC computer to accept Data (type 4) files sent using the TN XI module. I would like to be able to transfer TN spectral and image files to a Mac or PC but I am not sure of the file formats nor of a good communications program to use. If you have written software that you would be willing to share or purchased software that you no longer use and would like to sell, then please contact me.
Mark Germani, Ph.D. MicroMaterials Research, Inc. 136 Shore Drive, #200 Burr Ridge, IL 60521
Pippa Hawes wrote: ================================================================ I have a sample of dried porous silica that I would like to embed and thin section. Could anyone give me any advice on the best resin and protocol to use? The idea of using a methacrylate resin has been thrown about (the idea, not the resin - I can assure you), however I would be very grateful for any advice regarding any of the resins available. If this is a bit specific then reply off-line to the address below. ================================================================ We have always found that samples of this type (being run in our laboratory analytical services division), section quite nicely in most of the so-called "Epon® substitute" resins such as our own SPI-Pon™ Embedding Kit. We would expect that the "substitutes" offered by our main competitors would probably work just as well.
However, porous silica must be vacuum embedded, otherwise it can't be sectioned very well, if at all. We have never been able to get the section quality and stability with methacrylates or any system other than the "Epon" type kits. If one is working with "wet" silicas, and drying is not an option (as it sometimes is), then we have used our SPI-Chem™ Low Acid GMA as an embedding resin (which requires no dehydration).
The "Epon substitute" kits all come with recommended protocols, and while there is some variation between them, my guess is that any one protocol will work just as well with any of the other "substitutes". One advantage of the "Epon" systems is that the hardness can be varied quite easily, and often times the section quality can be improved by varying the hardness of the cured block.
Another protocol comment is that one can not use glass knives very easily (if at all) on this kind of sample, we use only (materials science) diamond knives.
Disclaimer: SPI Supplies, through our Structure Probe® laboratory services, perform these kinds of sample preps and TEM analysis for clients. We also are major suppliers of epoxy embedding resins and the SPI brand of materials science diamond knives.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} Hello, } I need to replace the SE scintillator on my SEM (JEOL 5900), and I was } considering using a YAP scintillator instead of the typical phosphorous } scintillator. Does anyone have any experience or suggestions on this?
About ten years ago I ran some tests for signal on several scintillator types: YAP, YAG, standard aluminum coated scintillator & "uncoated" scintillator. The result at that time were surprising.
Using a JEOL 35C SEM & running the same specimen current & PMT voltage with each scintillator the best signal was given by the original aluminum coated scintillator.
The second best was the "uncoated" scintillator & it was only 80% of the signal the standard aluminum coated scintillator. The YAP & YAG gave only 60% of the standard aluminum scintillator.
I was very disappointed as the YAP & YAG were significantly more than the standard scintillator. The conclusion that I came to was that the YAP & YAG would probably give more longevity & could justify their extra cost but they were not fit for high resolution. The YAP & YAG would probably be suitable for probe work.
Keep in mind this experiment was done about ten years ago. Perhaps the crystals have improved in recent years & the data may need a re-evaluation. I am sure that there is someone who would be willing to criticize my results (maybe in Northern Calif.?) but this the data that I received at the time.
Regards,
Earl Weltmer
----- Original Message ----- } From: "Johnson, Bradley R" {Bradley.Johnson-at-pnl.gov} To: "'Microscopy-at-MSA.Microscopy.com'" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, August 31, 2001 10:19 AM
I have followed this discussion with some interest. A similar situation exists in the field of mass spectroscopy.
At one time there were many mass spectrographs and mass spectrometers being used. Mass spectrographs were those instruments in which the detected ions were spread out along a focal plane and a significant portion of the mass range was collected simultaneously on a photographic plate. Mass spectrometers were those instruments in which the detected ions were moved across a detector slit (by varying magnetic field or ion accelerating potential) and collected/detected, one m/q at a time, by either a Faraday cup or electron multiplier after the slit, providing a readout on an electronic device (electrometer usually). Mass spectroscope or mass spectroscopy was used as the general term to include either type of instrument.
Mass spectrographs are rarely seen any more and even if they are used, the photographic plate would probably be replaced by some type of CCD detector providing electronic readout so the original clear distinction becomes blurred.
I'm sure that an analogous situation exists in optical spectroscopy.
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
I have followed this discussion with some interest. A similar situation exists in the field of mass spectroscopy.
At one time there were many mass spectrographs and mass spectrometers being used. Mass spectrographs were those instruments in which the detected ions were spread out along a focal plane and a significant portion of the mass range was collected simultaneously on a photographic plate. Mass spectrometers were those instruments in which the detected ions were moved across a detector slit (by varying magnetic field or ion accelerating potential) and collected/detected, one m/q at a time, by either a Faraday cup or electron multiplier after the slit, providing a readout on an electronic device (electrometer usually). Mass spectroscope or mass spectroscopy was used as the general term to include either type of instrument.
Mass spectrographs are rarely seen any more and even if they are used, the photographic plate would probably be replaced by some type of CCD detector providing electronic readout so the original clear distinction becomes blurred.
I'm sure that an analogous situation exists in optical spectroscopy.
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
You do not state the exact type of silica you are embedding. SO...
In general methacrylates, whether LR White or such, will shrink much more than an epoxy like one of the EPON CLONE formulations. If I recall 20-25% shrinkage is what you get with LR White and a lot of polymers. Epoxy formulas are more like 5%. This lower shrinkage is preferred by me so that I can argue to customers that I have not changed the microstructrure through severe shrinkage during the polymerization. LR White is faster (with an accelerator) curing, if you are in a hurry. The EPON clone stuff takes overnight curing in an oven at 60 to 70 C. It's no big deal because you just prepare the samples at the end of the day and start sectioning in the morning.
I have personnaly done precipitated silicas, arc silicas, and fumed silicas using Epon. Silica fume should be no problem. I do not vacuum embbed these materials. Ppt'd silicas have a surface area of about 154-250 meters squared per gram and the epon will wet almost every pore between the primary or ultimate particles without vacuum. Capillary forces are strong enough on our products and others to wet almost 100% of what is there. One of the components in my EPON kit foams under partial vacuum and that is another reason I don't use vacuum. You might have to use vacuum in your case. However, test each EPON kit component separately to see how much vacuum it can tolerate before foaming.
We have made various sizes of agglomerated spheres of these particles since the mid 70s as I recall. Anyway, they can be fragile but epon does not break up the microstructure. One can section throgh the 'micro-dust' on the surface of these spheres without loss of material.
As you can see, EPON works on a wide range of 'porous silica' samples. There must be a reason why EM people like it? It works on a wide range of materials, wets well, sticks fairly well to them, and sections well.
Eponate 12 releases from polyethelene molds, bottle caps, etc. quite well for 1-4 cycles of mold use with EPON, FYI.
Hope this helps.
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center Monroeville, PA 15146
Opinions given are my own and not those of PPG Industries.
} Dear all } } I have a sample of dried porous silica that I would like to embed and } thin section. Could anyone give me any advice on the best resin and } protocol to use? The idea of using a methacrylate resin has been thrown } about (the idea, not the resin - I can assure you), however I would be } very grateful for any advice regarding any of the resins available. If } this is a bit specific then reply off-line to the address below. } } Thanks } Pippa } ---------------------- } Pippa Hawes, EMU } School of Chemistry } University of Bristol } Cantocks Close } Bristol UK } BS8 1TS } Pippa.Hawes-at-bristol.ac.uk } } } }
Earl Weltmer and Brad Johnson wrote: ============================================================ } Hello, } I need to replace the SE scintillator on my SEM (JEOL 5900), and I was } considering using a YAP scintillator instead of the typical phosphorous } scintillator. Does anyone have any experience or suggestions on this? ++++Brad Johnson
About ten years ago I ran some tests for signal on several scintillator types: YAP, YAG, standard aluminum coated scintillator & "uncoated" scintillator. The result at that time were surprising.
Using a JEOL 35C SEM & running the same specimen current & PMT voltage with each scintillator the best signal was given by the original aluminum coated scintillator.
The second best was the "uncoated" scintillator & it was only 80% of the signal the standard aluminum coated scintillator. The YAP & YAG gave only 60% of the standard aluminum scintillator.
I was very disappointed as the YAP & YAG were significantly more than the standard scintillator. The conclusion that I came to was that the YAP & YAG would probably give more longevity & could justify their extra cost but they were not fit for high resolution. The YAP & YAG would probably be suitable for probe work.
Keep in mind this experiment was done about ten years ago. Perhaps the crystals have improved in recent years & the data may need a re-evaluation. I am sure that there is someone who would be willing to criticize my results (maybe in Northern Calif.?) but this the data that I received at the time. +++++ Earl Weltmer ============================================================ We have offered P-47, YAG and YAP scintillators for SEM applications for more then twenty years. Earl is correct that the P-47 performance, **but when first installed**, does probably give a superior performance. However , not all YAG and YAP crystals come out of the "same cookie cutter." From what we have been able to deduce, a high quality YAG or YAP single crystal scintillator will perform so close to that of the P-47 that one usually has difficulty detecting the difference. However, as everyone knows, the P-47 once installed, in terms of performance, goes only down. The only question is how fast it will go down. And that rate of deterioration of performance depends among other things on the type of work being done. For example, the higher x-ray fluxes associated with EDS (and especially WDS) tend to cause a more rapid deterioration.
Also, if Earl's experiment was done as described, and if a YAG was tested against P-47 but using the same PMT, then one would expect to find an inferior performance, because the PMT that is optimum for P47 is not the same one that would have been optimum for YAG. Now this might be less of a problem today than it would have been ten years ago, but if the PMT was not changed, then the right test (IMHO) was not the one being performed. To make the comparison, one would have had to have taken out the S11 style and replaced it with an S20 PMT. This point is further explained on our website URL http://www.2spi.com/catalog/scintill/spi-yag.html
When a YAG or YAP scintillator is installed, its performance level is constant and does not deteriorate. So while the comparison might suggest some superiority at the very beginning, in most instances that we have seen, or have been led to believe, it is not long that the SEM running on YAG or YAP in general, is operating at a higher level of performance. And the SEM is not subjected to the downtime associated with the need to change a P-47 powder scintillator from time to time.
For those operating at low KV in the BSE mode (and using YAG or YAP), and high magnifications, the image is clearly less noisy. A collection of ten different references from different laboratories around the world, which have been collected on URL http://www.2spi.com/catalog/scintill/sem-tem.html at least to some, pretty much document the superiority of the single crystal scintillators over the powder scintillators. I guess one could always try to do more studies, but anyone that is familiar with these publications finds their conclusions pretty persuasive.
We have generally recommended that the SPI single crystal scintillators offered many advantages over the more traditional P-47 scintillator. We have been under the impression that those who have made the switch have been pretty happy with their results, though we do hear from time to time of someone who would not agree with that statement. Of course, the performance is related to the nature of the doping of the crystal, and as I said, not all these crystals come out of the same cookie cutter. Not all YAG's and YAP's are created equal. So when making comparisons, and statements about their scintillators, it would be helpful for one to state the brand of their scintillator crystal. These crystals are certainly not a generic entity where all are the same irrespective of the source.
Disclaimer: SPI Supplies is a major supplier of scintillators for SEM applications including P-47's, YAGs and YAPs.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} } I have a sample of dried porous silica that I would like to embed and } thin section. Could anyone give me any advice on the best resin and } protocol to use? The idea of using a methacrylate resin has been thrown } about (the idea, not the resin - I can assure you), however I would be } very grateful for any advice regarding any of the resins available. If } this is a bit specific then reply off-line to the address below. } Pippa -
You'll probably do fine with LR White, Hard grade. Its low viscosity is a big help, but if the sample floats anyway you may need vacuum. If you've never used that resin, be cautious when selecting the embedding mold; it will dissolve some plastics & it won't harden in flat molds unless air is excluded. Gelatin capsules work.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I have followed this discussion with some interest. A similar situation exists in the field of mass spectroscopy.
At one time there were many mass spectrographs and mass spectrometers being used. Mass spectrographs were those instruments in which the detected ions were spread out along a focal plane and a significant portion of the mass range was collected simultaneously on a photographic plate. Mass spectrometers were those instruments in which the detected ions were moved across a detector slit (by varying magnetic field or ion accelerating potential) and collected/detected, one m/q at a time, by either a Faraday cup or electron multiplier after the slit, providing a readout on an electronic device (electrometer usually). Mass spectroscope or mass spectroscopy was used as the general term to include either type of instrument.
Mass spectrographs are rarely seen any more and even if they are used, the photographic plate would probably be replaced by some type of CCD detector providing electronic readout so the original clear distinction becomes blurred.
I'm sure that an analogous situation exists in optical spectroscopy.
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id AAA25972 for dist-Microscopy; Sun, 2 Sep 2001 00:06:16 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id AAA25969 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 2 Sep 2001 00:05:46 -0500 (CDT) Received: from gol-mas2.austar.net.au ([203.22.8.216]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id AAA25961 for {microscopy-at-sparc5.microscopy.com} ; Sun, 2 Sep 2001 00:05:34 -0500 (CDT) Received: from 150 (bds-228-109.tow.austar.net.au [203.21.228.109]) by gol-mas2.austar.net.au (Mirapoint) with SMTP id ADU82053; Sun, 2 Sep 2001 15:03:17 +1000 (EST) Received: by localhost with Microsoft MAPI; Sun, 2 Sep 2001 15:02:55 +1000 Message-ID: {01C133C0.5859C4A0.jim-at-proscitech.com} "jim-at-proscitech.com" {jim-at-proscitech.com} , "'Angela Klaus'" {avklaus-at-amnh.org} , "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com}
Reinhard Rachel also wrote on this back-channel and so I expect that you are right: I confused the early FE-SEM and FE-TEM. I saw the Siemens instrument in the Karlsruhe application lab in October 76. I was there for two other instruments but the large FE instrument was of course rather eye-catching. 25 years later I recall that it had a short, but large diameter column and so thought of it in the long-term as a FE-SEM. Hope that this was not a complete waste of time as these contributions have placed other FE instruments in a historical context. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, September 01, 2001 6:18 AM, Alan Nicholls [SMTP:nicholls-at-uic.edu] wrote: } Jim } } I think you mean FE-STEMs not FE-SEMs. The first VG HB5 STEM was delivered } in 1973 to University of London. } } Regards } } Alan } } At 09:55 AM 8/31/2001 +1000, Jim at Proscitech wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } In 1976 both, Siemens and Vacuum Generator marketed FESEMs. The Vacuum } } Generator unit was considerably cheaper and possibly better too. } } Incidentally, the Siemens unit became a fiasco and likely was a major reason } } for the company's board to abandon the building of electron microscopes a } } year } } or two later. Siemens had been the first commercial manufacturer of electron } } microscopes and built the best instruments in the 50th and early 60th. The } } Philips EM300 was the first serious challenge to Siemens. } } Michael Beer of Chicago published a great deal of FESEM developments in the } } early 70th. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ABN: 99 724 136 560 www.proscitech.com } } } } On Thursday, August 30, 2001 9:43 PM, Angela Klaus [SMTP:avklaus-at-amnh.org] } } wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi Listers... } } } } } } Does anyone know when the first cold field emission microscope became } } } commercially available? } } } } } } Thanks and best, } } } } } } Angela } } } } } } Angela V. Klaus } } } } } } Director, Interdepartmental Laboratories } } } American Museum of Natural History } } } Central Park West at 79th Street } } } New York, NY 10024 USA } } } } } } Email: avklaus-at-amnh.org } } } Tel: 212-769-5977 } } } } } Alan W Nicholls, PhD } Director of Research Service Facility (Electron Microscopy) } Research Resources Center - East (M/C 337) } Room 100 Science and Engineering South Building } The University of Illinois at Chicago } 845 West Taylor St } Chicago, IL 60607-7058 } } Tel: 312 996 1227 } Fax: 312 996 8091 } Office: Room 110 } } Web site www.rrc.uic.edu
Dear Gary, I will receive my Axiopklan only on next week - so I have still no experience and cannot help. With best wishes Jiri Kalvoda ----- Original Message ----- } From: Gary Gaugler {gary-at-gaugler.com} To: {jsharp-at-zeiss.com} Cc: MSA listserver {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 22, 2001 5:09 AM
Here, here! (or is it hear , hear?) Ken Converse Quality Images
Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listers, } } Nestor's little slip and his apology should make us realize how valuable Nestor and his service of the listserver is to us. I just want to say Thank You Nestor for all the hard work you do to keep us on-line and spam free. You help all of us to stay connected in a nice environment where we can ask our questions and communicate our ideas. } } Thank You Nestor! } } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax } } } }
-----Original Message----- } From: John A. Reffner [mailto:JAReffner-at-compuserve.com]
On Tuesday, October 2, 2001, Dr. Joseph I. Goldstein will receive the New York Microscopical Society's Ernst Abbe Memorial Award at a special symposium in Atlantic City, New Jersey. The Symposium is part of the Eastern Analytical Symposium & Exposition. The session starts at 9:00-am with the presentation of the Abbe Award. The program includes:
Joseph I. Goldstein Advances in Scanning Electron Microscopy and X-Ray Microanalysis
Charles E. Lyman Quantitative X-Ray Microanalysis in the Environmental SEM
David B. Williams Microbeam Analysis of Metallic Meteorites
Eric Lifshin Pushing the Limits of Spatial Resolution in X-Ray Microanalysis
Come and join the celebration Dean Goldstein's contributions to science of microscopy and educating so many microscopists.
The EAS meeting will be held in the Atlantic City Convention Center, October 1-4, 2001. For more information contact the EAS Homepage:( http://www.eas.org).
I have run into a file format problem between Semper for Windows(image processing software from Synoptics, UK running on Microsoft Windows 98) and EMS (image simulation software by Prof Stadelmann running on a Silicon Graphics workstation). The EMS suite of programs for image simulation has a routine se1 that writes the R type images (created by the im1 routine, in the present case) in Semper file format which is essentially a Fortran unformatted output. This file is to be read into Semper using the unformatted option of the read command. However I get a
Message: illegal structure for unformatted file ?129: File I/O error 6419 on unit 1 file 'd:\users\divakar\semper\images\rd1082.unf'
error on Semper for Windows and a similar message on Semper for DOS version 6.4. I would appreciate help / advise from members of this list in this regard. Is this because the binary code on Unix and DOS /Windows is different? Or is this a version problem? Does somebody have software / image file / disc file format details which can be used to inter-convert between these operating systems and / or hardware?
With Best Regards, ---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
We have an Electroscan ESEM E3, on which we have had different problems with the stage. In one case it was necessary to change some cables to fix the problem, but occasionally the stage won't move in the X-Y directions, and the problem is solved simply by changing the magnification on the microscope!! We haven't yet been able to come up with an explanation for this strange behaviour, but I hope for you, that your problems may be solved in this simple way.
Kind regards, Jesper ________________________________ Jesper Vejloe Carstensen Electron Microscopy & Microanalysis Risoe National Laboratory, Materials Research Department P.O. Box 49, DK-4000 Roskilde, DENMARK Phone: 46 77 57 76, Fax: 46 77 57 58 E-mail: jesper.v.carstensen-at-risoe.dk Web: www.risoe.dk/afm
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: 31. august 2001 18:24 To: Microscopy-at-sparc5.microscopy.com
Hello, I've been going through setting up an Electroscan ESEM E3 in our laboratory and vacuum problems aside, we now have a stage that doesn't want to move. Anyone ever come across stage problems and have some suggestions that I may try for this instrument?
The stage will actually move if you give it a gentle push along a gear arm, but it often locks up in the Z direction, and sometimes X/Y. Auto calibrate allows the stage to move in X/Y, but it never calibrates the Z direction. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Dear fellow list server members, We have a Si(Li) detector with Be/UT window attached to a SEM. Some time before I started my job here there was a vacuum fault where pump oil backstreamed into the SEM column -my understanding is that the Be window was in position at the time. We are now seeing low counts for light elements when used in thin window mode and quantitative data reflect this, even though the oil streamed in when the Be window was being used. The manufacturer has suggested the likely source is ice on the crystal (from figures below). This problem I am assured only started after the vacuum/oil problem which indicates oil is the likely problem. I include K/L line data collected from a copper sample at 10, 15, 20 kV with both the Be and UT windows in place. The counts represent the peak windows/ROI of the K and L lines. What do you make of these figures below? As I have no reference data or anything to compare with. I would appreciate any input or suggestions you have.
Beam Window Cu L Cu K K/L ratio 20 Be 47461 156947 3.3 20 utw 84033 110570 1.3 15 Be 18357 24336 1.3 15 utw 34930 18172 0.5 10 Be 15225 1070 0.07 10 utw 28005 880 0.03
Mant thanks.
Martin Roe Electron Microscopist Materials/Engineering Department Wolfson Building Nottingham University University Park Nottingham NG7 2RD tel +44 (0) 115 9513768 tel =44 (0) 115 9513871 fax +44 (0) 115 9513764 email: martin.roe-at-nottingham.ac.uk
Due to you didn't mention anything about live time, beam condition, i.e, beam current, beam alignment, working distance and ect.. so it is quite difficult to suggest anything about your results. However, it seems likely that at 15kv (Be) and 20k(utw) are OK. Normally, at 20kV, K/L ratio is close to 1, it depends on the condition of the detector.
Try Ni K/L to confirm that, K/L ratio would be about 1 at 20 kV.
Regards,
Paiboob Nuannin Dept of Physics Faculty of Science Prince of Songkla University Hatyai 90110 Thailand
On Mon, 3 Sep 2001, Martin Roe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Dear fellow list server members, } We have a Si(Li) detector with Be/UT window attached to a SEM. Some time before I started my job here there was a vacuum fault where pump oil backstreamed into the SEM column -my understanding is that the Be window was in position at the time. We are now seeing low counts for light elements when used in thin window mode and quantitative data reflect this, even though the oil streamed in when the Be window was being used. The manufacturer has suggested the likely source is ice on the crystal (from figures below). This problem I am assured only started after the vacuum/oil problem which indicates oil is the likely problem. I include K/L line data collected from a copper sample at 10, 15, 20 kV with both the Be and UT windows in place. The counts represent the peak windows/ROI of the K and L lines. } What do you make of these figures below? As I have no reference data or anything to compare with. I would appreciate any input or suggestions you have. } } Beam Window Cu L Cu K K/L ratio } 20 Be 47461 156947 3.3 } 20 utw 84033 110570 1.3 } 15 Be 18357 24336 1.3 } 15 utw 34930 18172 0.5 } 10 Be 15225 1070 0.07 } 10 utw 28005 880 0.03 } } Mant thanks. } } Martin Roe } Electron Microscopist } Materials/Engineering Department } Wolfson Building } Nottingham University } University Park } Nottingham } NG7 2RD } tel +44 (0) 115 9513768 } tel =44 (0) 115 9513871 } fax +44 (0) 115 9513764 } email: martin.roe-at-nottingham.ac.uk } }
We tried a stretch alignment of the column to verify the basic astigmatism.
In low mag, specially in the central and lower magnification range (between 50 and 250) we see a ?blade? cutting the field of view only without any condenser aperture inserted.
This blade is the objective aperture support.
I suppose it is a normal thing, but I like to be sure about it.
Is the same on your machine?
If it is so, please let me know.
Thanks a lot!
P.S. Thanks Nestor!
Marco Arienti
__________________________________________________________________ Abbonati a Tiscali! Con VoceViva puoi anche ascoltare ed inviare email al telefono. Chiama VoceViva all' 892 800 http://voceviva.tiscali.it
We tried a stretch alignment of the column to verify the basic astigmatism.
In low mag, specially in the central and lower magnification range (between 50 and 250) we see a "blade" cutting the field of view only without any condenser aperture inserted.
This blade is the objective aperture support.
I suppose it is a normal thing, but I like to be sure about it.
Regarding my recent experiences with my used Axioplan, I consider the issue to be closed. Zeiss sent a factory technician to my site and corrected the problem at no charge.
I have little experience with SEM and EDX detector performance, however, I have used a range of Be, thin window and windowless detectors on TEMs and there are a couple of comments I would like to make.
If the degradation in performance occurred directly after the oil backstreaming then I agree that oil is quite likely to be the cause but otherwise it is quite possibly ice on the crystal.
Has the detector liquid nitrogen always been kept well filled? If the internal 'cryo pump' starts to warm up then you could get icing on the crystal.
Why not condition the detector anyway? Certainly on my Oxford Instruments (ex Link) detectors it is easy to do and does not seem to degrade the performance at all. I regularly condition the windowless detector on my JEOL2010 TEM (about every 6 months) to get the light element performace back. However, I have not had to condition the SATW detector (on another TEM) in over two years so I guess the water comes from the microscope and not the detector.
Regards, Ron
On Mon, 03 Sep 2001 17:02:31 +0100 Martin Roe {Martin.Roe-at-nottingham.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Dear fellow list server members, } We have a Si(Li) detector with Be/UT window attached to a SEM. Some time before I started my job here there was a vacuum fault where pump oil backstreamed into the SEM column -my understanding is that the Be window was in position at the time. We are now seeing low counts for light elements when used in thin window mode and quantitative data reflect this, even though the oil streamed in when the Be window was being used. The manufacturer has suggested the likely source is ice on the crystal (from figures below). This problem I am assured only started after the vacuum/oil problem which indicates oil is the likely problem. I include K/L line data collected from a copper sample at 10, 15, 20 kV with both the Be and UT windows in place. The counts represent the peak windows/ROI of the K and L lines. } What do you make of these figures below? As I have no reference data or anything to compare with. I would appreciate any input or suggestions you have. } } Beam Window Cu L Cu K K/L ratio } 20 Be 47461 156947 3.3 } 20 utw 84033 110570 1.3 } 15 Be 18357 24336 1.3 } 15 utw 34930 18172 0.5 } 10 Be 15225 1070 0.07 } 10 utw 28005 880 0.03 } } Mant thanks. } } Martin Roe } Electron Microscopist } Materials/Engineering Department } Wolfson Building } Nottingham University } University Park } Nottingham } NG7 2RD } tel +44 (0) 115 9513768 } tel =44 (0) 115 9513871 } fax +44 (0) 115 9513764 } email: martin.roe-at-nottingham.ac.uk } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I'll second what Jesper says. Our E3 also occasionally does the "lock-up" thing but can always be freed up simply by changing the magnification. Why this should fix it is, I think, one of the great mysteries of the universe. We've also occasionally experienced slow movement of the stage in the XY direction. This can make acquiring a digital image a real problem at times. Usually you can stop this drift by changing the pressure in the chamber.....Great Mystery of the Universe #2.....
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia ----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, August 31, 2001 1:23 PM
Charles Garber in his recent posting wrote: } Not all YAG's and } YAP's are created equal. So when making comparisons, and statements about } their scintillators, it would be helpful for one to state the brand of their } scintillator crystal. These crystals are certainly not a generic entity } where all are the same irrespective of the source.
I am a bit puzzled by this. I was under the impression that all of the YAG and YAP scintillators offered for sale in this country originate from the same ultimate supplier in Europe. Has this changed? Or perhaps there are different grades of crystal?
Fred Schamber Aspex LLC
"Garber, Charles A." wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Earl Weltmer and Brad Johnson wrote: } ============================================================ } } Hello, } } I need to replace the SE scintillator on my SEM (JEOL 5900), and I was } } considering using a YAP scintillator instead of the typical phosphorous } } scintillator. Does anyone have any experience or suggestions on this? } ++++Brad Johnson } } About ten years ago I ran some tests for signal on several scintillator } types: YAP, YAG, standard aluminum coated scintillator & "uncoated" } scintillator. } The result at that time were surprising. } } Using a JEOL 35C SEM & running the same specimen current & PMT voltage with } each scintillator the best signal was given by the original aluminum coated } scintillator. } } The second best was the "uncoated" scintillator & it was only 80% of the } signal the standard aluminum coated scintillator. } The YAP & YAG gave only 60% of the standard aluminum scintillator. } } I was very disappointed as the YAP & YAG were significantly more than the } standard scintillator. } The conclusion that I came to was that the YAP & YAG would probably give } more longevity & could justify their extra cost but they were not fit for } high resolution. The YAP & YAG would probably be suitable for probe work. } } Keep in mind this experiment was done about ten years ago. Perhaps the } crystals have improved in recent years & the data may need a re-evaluation. } I am sure that there is someone who would be willing to criticize my results } (maybe in Northern Calif.?) but this the data that I received at the time. } +++++ Earl Weltmer } ============================================================ } We have offered P-47, YAG and YAP scintillators for SEM applications for } more then twenty years. Earl is correct that the P-47 performance, **but } when first installed**, does probably give a superior performance. However } , not all YAG and YAP crystals come out of the "same cookie cutter." From } what we have been able to deduce, a high quality YAG or YAP single crystal } scintillator will perform so close to that of the P-47 that one usually has } difficulty detecting the difference. However, as everyone knows, the P-47 } once installed, in terms of performance, goes only down. The only question } is how fast it will go down. And that rate of deterioration of performance } depends among other things on the type of work being done. For example, the } higher x-ray fluxes associated with EDS (and especially WDS) tend to cause a } more rapid deterioration. } } Also, if Earl's experiment was done as described, and if a YAG was tested } against P-47 but using the same PMT, then one would expect to find an } inferior performance, because the PMT that is optimum for P47 is not the } same one that would have been optimum for YAG. Now this might be less of a } problem today than it would have been ten years ago, but if the PMT was not } changed, then the right test (IMHO) was not the one being performed. To } make the comparison, one would have had to have taken out the S11 style and } replaced it with an S20 PMT. This point is further explained on our website } URL } http://www.2spi.com/catalog/scintill/spi-yag.html } } When a YAG or YAP scintillator is installed, its performance level is } constant and does not deteriorate. So while the comparison might suggest } some superiority at the very beginning, in most instances that we have seen, } or have been led to believe, it is not long that the SEM running on YAG or } YAP in general, is operating at a higher level of performance. And the SEM } is not subjected to the downtime associated with the need to change a P-47 } powder scintillator from time to time. } } For those operating at low KV in the BSE mode (and using YAG or YAP), and } high magnifications, the image is clearly less noisy. A collection of ten } different references from different laboratories around the world, which } have been collected on URL } http://www.2spi.com/catalog/scintill/sem-tem.html } at least to some, pretty much document the superiority of the single crystal } scintillators over the powder scintillators. I guess one could always try } to do more studies, but anyone that is familiar with these publications } finds their conclusions pretty persuasive. } } We have generally recommended that the SPI single crystal scintillators } offered many advantages over the more traditional P-47 scintillator. We } have been under the impression that those who have made the switch have been } pretty happy with their results, though we do hear from time to time of } someone who would not agree with that statement. Of course, the performance } is related to the nature of the doping of the crystal, and as I said, not } all these crystals come out of the same cookie cutter. Not all YAG's and } YAP's are created equal. So when making comparisons, and statements about } their scintillators, it would be helpful for one to state the brand of their } scintillator crystal. These crystals are certainly not a generic entity } where all are the same irrespective of the source. } } Disclaimer: SPI Supplies is a major supplier of scintillators for SEM } applications including P-47's, YAGs and YAPs. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ==================================================
The first VG HB5 FE-STEM was delivered in 1973 to University of London.
Both AEI and Seimens got into dedicated STEM shortly before they pulled out of the EM market in the early 70's. VG was able to attract a number of the main characters who had worked at AEI to form the core of the VG Microscopes unit.
The first electron microscope VG produced was a W thermal sourced SEM (Miniscan) for which they were awarded the Queens award for technological innovation in 1970. They then went on to produce the HB5 High Vacuum FE-STEM and HB50 High Vacuum FE-SEM. All of the latter were sold with Auger spectrometers as a Scanning Auger Microprobe.
Regards
Alan
At 09:55 AM 8/31/2001 +1000, Jim at Proscitech wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
On both the E3 and 2020 the stage movement with the Joy Stick is linked to magnification. The higher the mag the slower the stage speed. Above about 8000 X the stage motors are disabled and beam shift is active with the Joy Stick.
The apparent movement of the image should remain the same at all mags. If the mag is above about 8KX then the amount of image movement is limited by the beam shift. To continue searching the magnification has to be lowered.
For ElectroScan ESEM questions (E2,E3,Explorer, 2010 or 2020) world wide contact me directly at dsimpson-at-ma.feico.com For XL ESEM (XLFEG ESEM, XLLab6 ESEM or XLTMP ESEM in North America contact me directly at dsimpson-at-ma.feico.com For XL ESEM (XLFEG ESEM, XLLab6 ESEM or XLTMP ESEM for the rest of the world contact their local Service group.
Regards Derek Simpson ESEM Technical Support Manager FEI Company One Corporation Way #2 Peabody, MA 01960 E-mail: dsimpson-at-feico.com Voice: 1.978.538.6700 Fax: 1.978.531.9648
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Gordon,
We have an Electroscan ESEM E3, on which we have had different problems with the stage. In one case it was necessary to change some cables to fix the problem, but occasionally the stage won't move in the X-Y directions, and the problem is solved simply by changing the magnification on the microscope!! We haven't yet been able to come up with an explanation for this strange behaviour, but I hope for you, that your problems may be solved in this simple way.
Kind regards, Jesper ________________________________ Jesper Vejloe Carstensen Electron Microscopy & Microanalysis Risoe National Laboratory, Materials Research Department P.O. Box 49, DK-4000 Roskilde, DENMARK Phone: 46 77 57 76, Fax: 46 77 57 58 E-mail: jesper.v.carstensen-at-risoe.dk Web: www.risoe.dk/afm
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: 31. August 2001 18:24 To: Microscopy-at-sparc5.microscopy.com
Hello, I've been going through setting up an Electroscan ESEM E3 in our laboratory and vacuum problems aside, we now have a stage that doesn't want to move. Anyone ever come across stage problems and have some suggestions that I may try for this instrument?
The stage will actually move if you give it a gentle push along a gear arm, but it often locks up in the Z direction, and sometimes X/Y. Auto calibrate allows the stage to move in X/Y, but it never calibrates the Z direction. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
An alternative you may want to consider is the Polaron E6700 Evaporator, manufactured by Thermo VG Scientific. We are their distribution and service agent in the U.S. and would be happy to supply you with more information.
Best regards,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 mnesta-at-ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: K.N. Bozhilov [mailto:bozhilov-at-citrus.ucr.edu] Sent: Friday, August 31, 2001 12:27 PM To: microscopy-at-sparc5.microscopy.com
Hi All,
I would greatly appreciate if you could share your opinion about performance and reliability of the Bench Top Turbo III vacuum evaporator from Denton Vacuum, also alternative suggestions are more than welcome. We are looking currently to purchase a vacuum evaporator for general EM preparation capable of carbon and metal evaporation, glow discharge and thickness control. Cleanliness of the vacuum is our main concern and especially the fact that the Bench Top model does not have LN trap.
Thank you,
Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
We are parting with a used oil diffusion pump (w/ LN2 Trap) that has approx. 2 years wear and tear. The pump has been in storage for the last 10 years. The pump was manufactured by DAIA vacuum Engineering corp, model # DPF4Zs. The mounting flange is approx. 7.25 inches with an 8 hole bolt pattern. It was removed from a Hitachi S-4000 FESEM for replacement by a cleaner turbo model. If you are interested please contact me off-line. The pump is free but you must assume all shipping costs. Thanks, jr
I will hazard one guess being unfamiliar with either of the programs or formats that you describe. I suspect that there is a difference in how the two systems format text files. Under DOS and Windows, text lines normally end with a carriage return and line feed characters, ASCII codes 13 and 10. Under UNIX, lines often end with one or the other, but not both; I think carriage return is the norm.
I know that WS-FTP has an option for converting text files to the proper standard when transferring between the two different platforms. It might be able to help, but you will need to tell it that the file are text. It ordinarily runs in auto-detect mode and determines file type based on the extension. Unknown extensions default to binary format. Presuming you have to ship your files anyway, this might eliminate the need for a translator program.
There are also a number of utilities around that could give you a look at the binary codes for your file to see what characters are being used to indicate the end of the line. I have some I could recommend on the PC side if you don't already have one. They are relatively available on the shareware sites and could help verify what I have said. Some even serve as editors so you could make changes to one file manually to see if this is the source of your problem.
Warren
At 04:16 PM 9/3/2001 +0530, you wrote:
} I have run into a file format problem between Semper for Windows(image } processing software from Synoptics, UK running on Microsoft Windows 98) } and EMS (image simulation software by Prof Stadelmann running on a } Silicon Graphics workstation). The EMS suite of programs for image } simulation has a routine se1 that writes the R type images (created by } the im1 routine, in the present case) in Semper file format which is } essentially a Fortran unformatted output. This file is to be read into } Semper using the unformatted option of the read command. However I get } a } } Message: illegal structure for unformatted file } ?129: File I/O error 6419 on unit 1 file } 'd:\users\divakar\semper\images\rd1082.unf' } } error on Semper for Windows and a similar message on Semper for DOS } version 6.4. I would appreciate help / advise from members of this list } in this regard. Is this because the binary code on Unix and DOS } /Windows is different? Or is this a version problem? Does somebody have } software / image file / disc file format details which can be used to } inter-convert between these operating systems and / or hardware? } } With Best Regards, } ---- } Divakar R } Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research } Kalpakkam 603102, India } ----
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
It is Ladd's belief that the ideal vacuum evaporator should include a mechanical and a diffusion pump to get you to the 10(-7) range and should have a LN trap. The evaporation system should have a protective shield for cleanliness. If you are doing chromium evaporation then a turbo pump would be required.
John Arnott
Disclaimer: Ladd Research manufactures and sells vacuum evaporation systems. --
**** Please Note Our New Address, Fax and Phone Numbers ****
LADD RESEARCH 83 Holly Court Williston, VT 05495 USA
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K.N. Bozhilov wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi All, } } I would greatly appreciate if you could share your opinion about performance } and reliability of the Bench Top Turbo III vacuum evaporator from Denton } Vacuum, also alternative suggestions are more than welcome. We are looking } currently to purchase a vacuum evaporator for general EM preparation capable } of carbon and metal evaporation, glow discharge and thickness control. } Cleanliness of the vacuum is our main concern and especially the fact that } the Bench Top model does not have LN trap. } } Thank you, } } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } Tel 909 787 2998 } Fax 909 787 4324
It is possible to break the 8mm glass, but it can be very frustrating. If the users want wider knives, try to talk them into purchasing a diamond knife for thick sections. It is less expensive than diamonds for thin sectioning, and I think it is a good buy because of time saved making and changing knives and attaching water catchers. The only downside is that once there is a nick in the edge it is there until resharpening time.
I would like to prepare 0.05 M Cacodylate buffer pH 7.4, but do not have the recipe and protocol necessary for its preparation. Would it be possible to help me out?
Thanks in advance.
Patrícia Reis ____________________________ Escola Superior Biotecnologia Universidade Católica Portuguesa Rua Dr. António Bernardino Almeida 4200-072 Porto PORTUGAL Tel.: 225580044 Fax.: 225090351 e.mail: {mailto:pmreis-at-mail.esb.ucp.pt} pmreis-at-mail.esb.ucp.pt
I would like to attend a course on microscopy and imaging techniques which would include the latest developments in this field. Apart from the ones at MBL, Cold Spring Harbor and Jim Pawley's course in June, does anyone know of others. Being quite keen, I would prefer something this winter but any suggestions would be highly appreciated.
Thank you Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
Dear All, A colleague recently inherited an IEC Minot Rotary Microtome. Does anyone have a manual or parts? It is missing the speciman holder, mounting bar for knife height adjustment and means of measuring knife angle.
Thanks in advance, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax ************************************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
Could also be a processor related problem. There is a difference in the way the Intel and Motorola processors store information. One of the two writes the LSB (least significant byte) first, the other the MSB (most significant byte). The best bet would probably be to use a format that can be read by both. TIF, for example, can be normally read by both as the file itself contains information about the byte order.
Can you transform your images into TIF?
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Tuesday, September 04, 2001 10:31 AM To: divakar-at-igcar.ernet.in Cc: Microscopy-at-sparc5.microscopy.com
I will hazard one guess being unfamiliar with either of the programs or formats that you describe. I suspect that there is a difference in how the two systems format text files. Under DOS and Windows, text lines normally end with a carriage return and line feed characters, ASCII codes 13 and 10. Under UNIX, lines often end with one or the other, but not both; I think carriage return is the norm.
I know that WS-FTP has an option for converting text files to the proper standard when transferring between the two different platforms. It might be able to help, but you will need to tell it that the file are text. It ordinarily runs in auto-detect mode and determines file type based on the extension. Unknown extensions default to binary format. Presuming you have to ship your files anyway, this might eliminate the need for a translator program.
There are also a number of utilities around that could give you a look at the binary codes for your file to see what characters are being used to indicate the end of the line. I have some I could recommend on the PC side if you don't already have one. They are relatively available on the shareware sites and could help verify what I have said. Some even serve as editors so you could make changes to one file manually to see if this is the source of your problem.
Warren
At 04:16 PM 9/3/2001 +0530, you wrote:
} I have run into a file format problem between Semper for Windows(image } processing software from Synoptics, UK running on Microsoft Windows 98) } and EMS (image simulation software by Prof Stadelmann running on a } Silicon Graphics workstation). The EMS suite of programs for image } simulation has a routine se1 that writes the R type images (created by } the im1 routine, in the present case) in Semper file format which is } essentially a Fortran unformatted output. This file is to be read into } Semper using the unformatted option of the read command. However I get } a } } Message: illegal structure for unformatted file } ?129: File I/O error 6419 on unit 1 file } 'd:\users\divakar\semper\images\rd1082.unf' } } error on Semper for Windows and a similar message on Semper for DOS } version 6.4. I would appreciate help / advise from members of this list } in this regard. Is this because the binary code on Unix and DOS } /Windows is different? Or is this a version problem? Does somebody have } software / image file / disc file format details which can be used to } inter-convert between these operating systems and / or hardware? } } With Best Regards, } ---- } Divakar R } Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research } Kalpakkam 603102, India } ----
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Dear lister? Our Narishige WR-90 micromanipulator does not work properly because the the water inside the system has leaked out slowly. Does anyone in the list know how to refill it? Any suggestion is highly appreciated. Thanks, Long ----------------------------------- Miao, Long Dept of Biological Science 334 Bio. Unit1 Florida State University Tallahassee, FL32306-4370 email: lmiao-at-bio.fsu.edu Voice: (850)644-9817 FAX : (850)644-0481
Dear Listers, If any of you have removed a jammed film cassette from a JEOL 1200EXII, I would like to know how you dislodged it. We will eventually have a service technician to help but I cannot figure out how to remove the plate. I did manage to remove the film and yes the plate was inserted right side up. Thanks in advance. Rosemary
A one day short course on "Digital Image Capture and Management in Light Microscopy" by Dr. Mary McCann & Dr. John McCann is being offered on Sunday September 30,2001 at the Eastern Analytical Symposium and Exhbition in Atlantic City, NJ.
Don O'Leary
-----Original Message----- } From: Judy Trogadis [mailto:TrogadisJ-at-smh.toronto.on.ca] Sent: Tuesday, September 04, 2001 9:13 AM To: Microscopy-at-sparc5.microscopy.com Cc: Judy Trogadis
Fellow Microscopists:
I would like to attend a course on microscopy and imaging techniques which would include the latest developments in this field. Apart from the ones at MBL, Cold Spring Harbor and Jim Pawley's course in June, does anyone know of others. Being quite keen, I would prefer something this winter but any suggestions would be highly appreciated.
Thank you Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
I have experience many jams. Generally occurring with old age. Anyway, JEOL camera mechanisms are wonderful mechanical beasts. Two pushing nails in the mechanism catch the plates. If you look at the plates, on one side there are two small rectangles, which are cut out. The nails use these to capture the plates and push them through the mechanism. These nails, together with the plates wear down and eventually sometimes they miss and do not mate. Generally jamming in the open slot where the plates come out of the box.
So, to dislodge a jam there are several solutions:
1) Waggle the mechanisms in the hope it clears - not a good choice. 2) Make a piece of stiff plastic sheet the width of a plate and the length around 15". Try to insert the sheet above the canisters but below the top mechanisms so as to lift the nails free of the canisters. You can see the nails when you look into the camera above the boxes on both sides. They look like small arms with a piece of copper alloy on the end. Pull camera out if nails are free. 3) Remove the whole camera mechanism. Simply remove the bolts from below the camera chamber (6 I think), and then pull the camera mechanism out in one piece. The jam can then be freed by rotating the mechanism a wee bit.
If you have frequent jams, consider replacing the pushing nails (first) then the plates. Also a good idea to replace the Teflon sheet bearings, but get a JEOL engineer to do this otherwise the position of the mechanism could be lost.
Good luck,
Keith. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
I have gotten ahold of a Zeiss camera lucida drawing tube. It has a mirror at an angle that allows one to draw underneath the inclined eye tube. Works for a Zeiss Standard 16. I am getting the hang of it a bit. However, I have at least one question:
What is the collar for (referred to as "ring clamp" below)? It is somewhat eccentric, but I cannot imagine any purpose for it. The device doesn't fit over it, but nearly does. It *is* useful for attaching it to a wider eyepiece tube.
This plastic collar fits snugly inside the eyepiece tube of an Olympus SZH; this is good. Also, it fits over the end of the standard 16 eye tube, with a barrier that doesn't allow it to slip down over the tube. i have some documentation for a similar drawing tube, but there is a slight difference in the picture (there is a knurled knob at the point where the mirror/prism fits over the eyepiece. Mine doesn't have that, it fits snug). Here is the relevant section from the documentation for the pictured device:
1. Focus microscope.
2. Remove eyepiece and slip ring clamp on to tube. Replace eyepiece and fix with ring clamp. [I referred to the "ring clamp" as a collar.]
3. Clamp drawing prism on the eyepiece. [I assume this refers to the knurled screw that isn't present on my device---mine doesn't need to be clamped on.]
} From reading the description, though, I don't know what it is for. Is it a spacer to move the eyepiece up higher? Which Zeiss is it really designed for?
Also, I would greatly appreciate any tips. The tips I found in MICSCAPE were helpful. Can I initiate some correspondence with someone who has some experience with one of these fantastic devices?
Alan Davis Marianas High School
-- adavis-at-saipan.com 1-670-235-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
Rosemary, I believe that the JEOL 1200 camera chamber is the same as the JEOL 100CX. You can slide out the film tray by making two metal strips measuring about 1" to 2" wide and about 10" to 12" long. Bend one end of each strip in the shape of an allen wrench to use as a sort of handle. These metal strips are used to raise up the two little arms that are locking up the film tray. While sitting on the floor so that the open camera chamber is at eye level and using a bright light you can look into the top area of the camera chamber just above where the film boxes are located. On each side and above the film boxes you will see these arms sticking down. You can use the metal strips to release these arms be sliding the metal strips straight in. Be sure that the film tray is all the way in so that there is no force on these arms holding them down. They are spring loaded and should raise up with only gently pushing on the metal strips. Be careful and do not apply a lot of force. You should be able to see these arms raise when you push in the metal strip. When both arms are raised the drawer should pull out easily. Again, don't use force. Hope that this helps.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
Systran Federal Corporation in Dayton, Ohio, is managing an "on-site" Contract for the Materials and Manufacturing Directorate (ML), Air Force Research Laboratory (AFRL), at Wright Patterson Air Force Base, in Dayton, Ohio. Candidates are sought for a research position at AFRL/ML, and this position entails research and development on understanding surface forces and surface chemistry of MEMS devices and MEMS materials:
a) study surface forces of MEMS using scanning probe microscopes and special adhesion/friction testers as a function of environment (humidity, temperature, vacuum); develop an understanding of how surface forces, and hence friction and wear may be controlled using surface treatments including monolayers and solid coatings b) study surface chemistry, tribochemistry, and coating microstructure to elucidate the mechanisms controlling friction and wear in MEMS systems under extreme environments c) research lubricant transport behavior and dynamics in MEMS systems.
The successful candidate will have knowledge/experience of MEMS devices, surface chemistry, surface forces and tribology. Expertise in surface chemical probes such as scanning Auger microscopy, X-ray photoelectron spectroscopy, FTIR microscopy / grazing angle FTIR, micro Raman, and SEM/EDS are highly desirable. The candidate will use apparatus to study surface forces at the nano scale (scanning probe microscopes, special adhesion testers, and nano/micro friction measurement devices). Working knowledge of monolayer and solid coatings is also highly desirable. The focus of the research effort is to understand the chemistry and physics controlling friction and wear and to use that understanding to develop effective technologies leading to reliable MEMS operation in extreme environments. Analytical instrumentation is currently available, but the successful candidate must be capable of modifying instrumentation as part of the research program.
The advertised position is a full-time job with complete benefits (paid vacation, health and dental coverage, 401K, etc.). Interested candidates should respond to this advertisement by sending their resumes along with a listing of technical publications to the contact person listed below.
Dr. V. ("Nagu") Nagarajan V.P. & Program Manager Systran Federal Corporation 4027 Colonel Glenn Highway, Suite 210 Dayton, OH 45431-1672
We have an SEM application which requires an array of samples to be loaded into the microscope. For producing the array, we need to affix sticky tabs to an aluminum block (48 tabs in a 6 x 8 arrangement on a block about 3 inches by 4 inches).
Presently, we are producing these arrays by hand, but it would be much more efficient to produce the arrays in this form by simply pressing a sheet onto the block, then peel it off to give us all of the 48 tabs properly located on the plate in one shot.
Does anyone have a name of the manufacturer(s) of carbon sticky tabs, so we can contact them about this. Our supplier is not willing to provide this information.
On our search for high N.A. water-dipping/immersion objectives for 160 mm-corrected microscopes we came across some lenses made by a Russian company called 'LOMO Optics' (www.lomooptics.com), which we tried out on our Nikon Optiphot 2 two-photon scope, and on a Nikon Measurescope MM-11 (bottom illumination). Their performance is quite astonishing. And since we had to look for quite a while to find such lenses, we wanted to let the rest of the community know that you do not have to give away your old 160 mm-corrected scope yet. Prices are quite reasonable too. At LOMO they are willing to let you try out the lenses for yourself. But do not be surprised to get a phone call every day during that time. (And do not forget to negotiate!)
[1] Type EAF-A30-1, TT079, 30x water-immersion, N.A. 0.9, w.d. 1.3 mm, 0.17 mm coverslip correction (performance does not seem to suffer without coverslip), built-in iris, outer diameter: 0.545" at the tip for about 0.2", and 0.825" thereafter (the objective has a cap of that diameter, which (at least in our case) could be removed, giving an even smaller outer diameter of 0.710"), overall length when installed: ~1.050".
[2] Type 96001, 70x water-dipping (apparently no coverslip correction)., N.A. 1.23, w.d. 220 µm, outer diameter -at- the tip: 0.426" to 0.585" conical for about 0.2", 0.800" above,overall length when installed: 1.26".
[3] Type 83127, 85x water-immersion., N.A. 1.0, w.d. between 200 and 230 µm (depending on the coverslip correction setting, which ranges from 100 µm to 200 µm), outer diameter -at- the tip: 0.733" for about 0.2", 0.825" above, overall length when installed: 1.26".
[4] Apparently they also sell a 100x water-immersion lens by now, which we have not seen yet.
With best regards,
Axel
__________________________________
Axel Blau California Institute of Technology Division of Biology 156-29 1200 E. California Blvd. Pasadena, CA 91125
I was at a talk where the carbon sticky dots were first presented. I wish I could remember who was speaking. I know that SPI has the exclusive distribution rights to them. I believe that they were developed by a group at Dupont. I would check the literature and see if they published. Try back issues of the meeting abstracts. I think that the talk that I went to was about 10 years ago.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Wednesday, September 05, 2001 10:17 AM To: 'Microscopy-at-sparc5.microscopy.com' Cc: Kuechl, Dorothy
Dear Listers,
We have an SEM application which requires an array of samples to be loaded into the microscope. For producing the array, we need to affix sticky tabs to an aluminum block (48 tabs in a 6 x 8 arrangement on a block about 3 inches by 4 inches).
Presently, we are producing these arrays by hand, but it would be much more efficient to produce the arrays in this form by simply pressing a sheet onto the block, then peel it off to give us all of the 48 tabs properly located on the plate in one shot.
Does anyone have a name of the manufacturer(s) of carbon sticky tabs, so we can contact them about this. Our supplier is not willing to provide this information.
I can't help you get it out, but just wanted to warn you that it might be possible for you to get your fingers crushed or severed if you're fishing around in the chamber and the jam suddenly frees itself. BE CAREFUL. I'm not sure if this is the case with your model, but it was on an older JEOL that we had. I'd wait for the tech or at least get his advice before reaching in.
We had some that would get caught, and the only thing to do with them after they were freed was to toss them. Don't try to straighten them.
Sara
On Tue, 4 Sep 2001, Rosemary Walsh wrote:
} Date: Tue, 4 Sep 2001 20:23:07 -0500 } From: Rosemary Walsh {rw9-at-psu.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: removing jammed TEM film cassettes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } If any of you have removed a jammed film cassette from a JEOL } 1200EXII, I } would like to know how you dislodged it. We will eventually have a service } technician to help but I cannot figure out how to remove the plate. I did } manage to remove the film and yes the plate was inserted right side up. } Thanks in advance. } Rosemary } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
The jamming of the plate drive mechanisms in electron microscopes is nothing new. I remember back in the 1960s when I was using our two JEM-6A TEMs in my classes on electron Microscopy. With an average enrollment of 25 to 30 students per term, coming from all disciplins across campus, and having all degrees of mechanical aptitude, plate jams were not an uncommon event. The easiest way to clear up a really bad plate jam in the JEM-6As was to reach inside the plate chamber and wiggle the plate carrier around a bit until things worked loose. Unfortunately, however, the chamber opening was too small for my hands to get in deep enough to take care of a bad jam, and so on several occasions I had to enlist the help of my 5-year-old son, who eventually became quite expert at taking care of the problem. Times do change, but digital cameras have their problems too!
Good luck, and keep smiling, WCB -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Does anyone know where I could purchase a battery powered vibrating engraving tool? We have the corded type but I need a portable one to use out on a factory floor. Vendor replies welcome
Paul D. Nolan Electron Optics
Alcan International Limited Kingston Research and Development Centre P.O.Box 8400, 945 Princess Street Kingston, Ontario K7L 5L9
Hi, all I would like an advice. Can anyone suggest a better type of dishes to use with glutheraldehyde and/or osmium tetroxide during fixation time. Which bottles are better to store these fixatives? Many thanks.
Patricia ____________________________ Patrícia João Reis (PhD student) Escola Superior Biotecnologia Universidade Católica Portuguesa Rua Dr. António Bernardino Almeida 4200-072 Porto PORTUGAL Tel.: 225580044 Fax.: 225090351 e.mail: {mailto:pmreis-at-mail.esb.ucp.pt} pmreis-at-mail.esb.ucp.pt
"Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed by ETOH dehydration, CPD, mounting and SEM viewing.
O.k., we give up, searched numberous books, papers, catalogs, web sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. "Champy's Solution"? Please?
Thank you much!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
} We had some that would get caught, and the only thing to do with them } after they were freed was to toss them. Don't try to straighten them. }
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Anyone out there knows how to run up an X-ray film (exposed as well as unexposed) for TEM????? Neelima Shah.............. Biomedical Imaging Core Facility Uni of Pennsylvania Philadelphia, Pa.
LOMO (Leningrad's Opto-Mechanics Consortium) is well known manufacturer of the fine optics in the Russia. Their microscopes are cheap (relatively) and very suitable for students etc. LOMO also known as an manufacturer of the biggest single-piece 6 meters diameter glass mirror for the telescope situated in Caucasus mountains. That mirror was a biggest one in the world at the time of inventing. I believe, they did also first full-quartz-optic microscope for UV investigations and their MB-1/2 fluorescence microscopes with "opak-illumination" were great.
I don't have any financial interest in this company.
Sergey Ryazantsev
At 10:03 AM 9/5/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Ah, the wonders of a Google search! I found a wonderful histo manual at
http://131.229.114.77/histo/Manual.pdf
The recipe is given on p. 17 of this pdf but it is in code - the author lists standard histology stock solutions and then gives the recipes for a variety of fixatives by coding them from the stock solutions. Champy's is: "35G; 20K; 12M; 24Q(9)" which means: 35 ml of 1% chromic acid (aq.), 20 ml of 2% OsO4 (aq.), 12 ml of sat. potassium dichromate (9 g/ 100ml, aq.), 24 ml of distilled water, make up just before use. Sounds like nasty stuff.
I'm glad you asked - I'd have never found this pdf otherwise! (It is 113 pages and I haven't been able to figure out from where it originates - some histology course). If anyone knows who wrote it, I'd like to know!
Tamara
On Wed, 5 Sep 2001, Richard Edelmann wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
I think I saw some at "Fredericks" out here on the West Coast. Contact me offline for the number.
Earl
----- Original Message ----- } From: {"Paul.Nolan-at-alcan.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, September 05, 2001 12:42 PM
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Scott Walck wrote: =================================================== I was at a talk where the carbon sticky dots were first presented. I wish I could remember who was speaking. I know that SPI has the exclusive distribution rights to them. I believe that they were developed by a group at Dupont. I would check the literature and see if they published. Try back issues of the meeting abstracts. I think that the talk that I went to was about 10 years ago. =================================================== Let me shed some light on this: SPI Supplies was (and is) the exclusive licensee for the manufacturing and distribution of Tacky Dot™ Slides. The patent holder was DuPont and the inventor was Dr. Alan Cairncross, still at DuPont. I am guessing, but the paper you heard presented (at an MSA meeting ) which could have been about ten years ago (how time flies), probably was presented by Dr. Cairncross (and other authors all at DuPont). Full technical details are disclosed in US Patent # 5,356,751.
These should not be confused however with what are often times called by others as "carbon sticky dots" or "double sided conductive adhesive tabs". These are offered in various forms by SPI as well as the other main worldwide distributors of consumable supplies for SEM applications.
The original poster (Wharton Sinkler) asked about adhesive "tabs" but did not specify size. The largest dot size of the Tacky Dot Slides manufactured by SPI Supplies is 300 µm in an orthogonal array on 2000 µm centers. I think that Wharton was in essence asking about the smallest "carbon tab" or "disc" that is available but the smallest that I know of are are 9 mm and there are technical reasons why one could not go too much smaller than that (in our opinion).
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
from "Notes on microscopical technique for zoologists" CFA Pantin Cambridge Univ. Press 1962
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
The business of un-jamming JEOL plate camera doesn't have to be that gory. Just remove a fuse located a couple inches left of the camera door (make sure that little red light next to the fuse turns on), before reaching inside, and no fingers will be shed.
Assuming that the camera mechanism is not damaged- simplest thing to do to avoid jams- do not load film boxes to a full capacity(say, load 5 or 10 holders less), make sure that film holders are in perfect shape - ditto on Sara's comment, toss the damaged holders- and that the film holders are loaded correctly. Also, little Teflon rollers on the sides of the plate boxes must rotate freely (at a slight touch, no friction) and must have spotless surface (no defects, scratches, etc.). Replacement rollers are available from JEOL. If you are going to unscrew 6 screws underneath the camera compartment in order to pull the mechanism out (as was suggested in one of the previous messages)- beware that those screws are separating camera vacuum from the atmosphere. Make sure that the o-rings, screws, and seats are clean before replacing the screws, to avoid vacuum leak.
If the mechanism is damaged- repair options will depend on you mechanical aptitude. You are welcome to contact me off-line for further advice, yet I agree that your best bet is to contact JEOL service representative.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Marie E. Cantino {cantino-at-uconnvm.uconn.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Reference: 'The Microtomist's Formulary and Guide' by Peter Gray. Blakiston Company, Inc., New York/Toronto. 1954:192 (794 pp.)
Cheers! Ken
-------------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
On Wed, 5 Sep 2001, Richard Edelmann wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." }
Does anyone know what has happened to the journal Scanning Microscopy?
The latest part we have is Volume 10 pt.4 1998 and our supplier cannot trace the publisher etc. all mail being marked return to sender. Has the journal ceased publishing?
Chris Jeffree
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Actually, the size of the tabs is not the issue. Sorry about the confusion.
What I was asking about are indeed double-stick carbon tabs about 1 cm diameter. I'd like to evenly space 48 of them on an aluminum plate, and am looking for a less tedious way than by hand.
An alternative would be to simply cover the entire plate with a double-stick conductive carbon layer, then depsoit the samples in the proper locations. This may be cheaper than getting something custom made. Does anyone know who might offer unperforated double stick conductive carbon material in large dimensions (about 3 x 4 inches)?
Thanks, Wharton
} -----Original Message----- } From: Garber, Charles A. [SMTP:cgarber-at-2spi.com] } Sent: Wednesday, September 05, 2001 9:20 PM } To: MICROSCOPY BB } Subject: Sticky dots and Tacky Dots } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Scott Walck wrote: } =================================================== } I was at a talk where the carbon sticky dots were first presented. I wish } I } could remember who was speaking. I know that SPI has the exclusive } distribution rights to them. I believe that they were developed by a } group } at Dupont. I would check the literature and see if they published. Try } back issues of the meeting abstracts. I think that the talk that I went } to } was about 10 years ago. } =================================================== } Let me shed some light on this: SPI Supplies was (and is) the exclusive } licensee for the manufacturing and distribution of Tacky Dot(tm) Slides. } The } patent holder was DuPont and the inventor was Dr. Alan Cairncross, still } at } DuPont. I am guessing, but the paper you heard presented (at an MSA } meeting } ) which could have been about ten years ago (how time flies), probably was } presented by Dr. Cairncross (and other authors all at DuPont). Full } technical details are disclosed in US Patent # 5,356,751. } } These should not be confused however with what are often times called by } others as "carbon sticky dots" or "double sided conductive adhesive tabs". } } These are offered in various forms by SPI as well as the other main } worldwide distributors of consumable supplies for SEM applications. } } The original poster (Wharton Sinkler) asked about adhesive "tabs" but did } not specify size. The largest dot size of the Tacky Dot Slides } manufactured } by SPI Supplies is 300 µm in an orthogonal array on 2000 µm centers. I } think that Wharton was in essence asking about the smallest "carbon tab" } or } "disc" that is available but the smallest that I know of are are 9 mm and } there are technical reasons why one could not go too much smaller than } that } (in our opinion). } } Chuck } } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== }
Thank you for all who replied we now have a recipe to try out.
Champy's:
=========
1% chromic acid 7 ml 3% K2Cr2O7 7 ml 2% OsO4 4 ml
from "Notes on microscopical technique for zoologists" CFA Pantin Cambridge Univ. Press 1962
========
"[3% aqueous potassium dichromate; 1% aqueous chromic acid; 2% aqueous solution of osmic acid (osmium tetroxide); (2:2:1)]" Page 804-805 Samuel Thompson's "Selected Histochemical and Histopathological Methods", 1966.
==========
"35G; 20K; 12M; 24Q(9)" which means: 35 ml of 1% chromic acid (aq.), 20 ml of 2% OsO4 (aq.), 12 ml of sat. potassium dichromate (9 g/ 100ml, aq.), 24ml of distilled water, make up just before use. Sounds like nasty stuff.
===========
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Does anyone out there know: a) where to buy this stuff? b) How to make this stuff? or c) where I can find any articles, mentions or instructions on the use of this stuff? (other than the Fisheries Science article that comes up in Google) I've been told that it's a useful marker for the aragonite polymorph of CaCO3. (Thanks, John Hunt at CCMR!) Any feedback, however oblique, would be appreciated.
Jeff
-- Jeff Hedlesky Aladdin Companies 151 N. Main, Suite 700 Wichita, KS 67202
It's been a while since I've been on the list, and since I've been working as a microscopist.
If I image a sample in the SEM with the sample tilted, then want to make measurements from the images, what is the equation to use to correct for the tilt? I know it involves the cosine of the tilt angle, but beyond that, I'm drawing a blank.
Thanks for your assistance.
Nancy Zjaba, Technical Staff Alfalight Inc. 1832 Wright Street Madison, WI 53704 (608) 240-4875 nzjaba-at-alfalight.com
Thanks to the two of you for the information. The website -at- www.comet.net/gek/ is very interesting, and I did not know that there was some other distributor in the US. I have to agree that the quality of the lenses we got is outstanding for the price we paid.
With best regards,
Axel
----- Original Message ----- } From: Jeff Hedlesky To: Sergey Ryazantsev Cc: Axel Blau ; Microscopy-at-sparc5.microscopy.com Sent: Thursday, September 06, 2001 6:56 AM
For a flat sample with tilted to an angle, theta, the image perpendicular to the tilt axis is foreshortened by cos(theta). Take your measurement on the tilted image and divide by cos(theta).
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Nancy Zjaba [mailto:nzjaba-at-alfalight.com] Sent: Thursday, September 06, 2001 12:18 PM To: 'microscopy-at-msa.microscopy.com'
Hello to the list.
It's been a while since I've been on the list, and since I've been working as a microscopist.
If I image a sample in the SEM with the sample tilted, then want to make measurements from the images, what is the equation to use to correct for the tilt? I know it involves the cosine of the tilt angle, but beyond that, I'm drawing a blank.
Thanks for your assistance.
Nancy Zjaba, Technical Staff Alfalight Inc. 1832 Wright Street Madison, WI 53704 (608) 240-4875 nzjaba-at-alfalight.com
After tilting, the feature you want to measure will form the hypotenuse of a triangle. The horizontal projection you see will be the side of the triangle adjacent to the tilt angle. One side of the feature will be elevated with respect to the other and that amount will be the length of the side opposite the tilt angle.
The cosine of an angle is the ratio of the length of the adjacent side to the length of the hypotenuse. Therefore, you would use the following formula.
hypotenuse = adjacent/cosine, or
real length - observed length/cosine(tilt angle).
At 11:18 AM 9/6/2001 -0500, you wrote:
} Hello to the list. } } It's been a while since I've been on the list, and since I've been working } as a microscopist. } } If I image a sample in the SEM with the sample tilted, then want to make } measurements from the images, what is the equation to use to correct for the } tilt? I know it involves the cosine of the tilt angle, but beyond that, I'm } drawing a blank. } } Thanks for your assistance. } } Nancy Zjaba, Technical Staff } Alfalight Inc. } 1832 Wright Street } Madison, WI 53704 } (608) 240-4875 } nzjaba-at-alfalight.com
I was working with JEOL microscopes (100B, 100C, 100CX, 1200EX) for a few decades. Over all this time I do remember just a few accidents related to the camera mechanics in the microscopes. In all that cases the "human factor" was involved and usually the problem was improperly loaded into cassette the piece of the film. If film is not in place, sometime it cover one of the square slots on the cassette and "fork" could not catch the cassette properly. In this case the forks usually stick in the gap between two set of rails in the camera mechanism. As it was mentioned in the most replies, the solution is to move up those forks out of the gap. Usually I am doing so using flexible metal ruler with fuse OFF (great comment Vitaly). The bad things about JEOL camera design that it has extremely powerful motor with huge gear which is able to bent and ruin all rails in the camera if working improperly. Magically, it was never happens with my microscopes, but when I look inside the camera and see that huge gears I am worrying anyway.
Sergey
At 12:12 AM 9/6/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Wharton Sinkler wrote: ================================================= Actually, the size of the tabs is not the issue. Sorry about the confusion.
What I was asking about are indeed double-stick carbon tabs about 1 cm diameter. I'd like to evenly space 48 of them on an aluminum plate, and am looking for a less tedious way than by hand.
An alternative would be to simply cover the entire plate with a double-stick conductive carbon layer, then depsoit the samples in the proper locations. This may be cheaper than getting something custom made. Does anyone know who might offer unperforated double stick conductive carbon material in large dimensions (about 3 x 4 inches)? ================================================== We at SPI Supplies offer as a standard off-the-shelf product a sheet that is 500mm x 500 mm. You could cut out from that standard size sheet whatever size you might require. This is described on URL http://www.2spi.com/catalog/spec_prep/cond_adhes-sheets.html
This is the "master material" from which the double sided conductive carbon adhesive discs are fabricated.
We have heard of another customer who has made a template in the orthogonal arrangement required and then by hand is affixing (on large sheets) what they want to attach through the use of the template, which is suspended slightly above the adhesive and never actually contacts the adhesive. That would be the cheapest of the various alternatives.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle with a Teflon lined screw cap. The bottle is then held in the refrigerator in a small plastic jar (available from all lab distributors - mine from Fisher or S/P) with a screw cap . In this way, the plastic becomes osmicated NOT the refrigerator. I have done this for years with no untoward results.
I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or plastic. Osmication is done in glass vials with Teflon lined screw caps using a minimum of OsO4 to do the job.
I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a Teflon lined cap. I then make up an appropriate amount of fixative using the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept in a plastic jar in the refrigerator.
I find that double containers are safe and reduce contamination considerably.
Hope this helps.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
Hello everyone, I have a researcher here that is having some difficulty with getting good histology at the EM level. She is fixing Drosophlia brains with 4%para/0.5% glut in phosphate buffer pH7.2, no osmium post fix, ethanol dehaydration, propylene oxide as a transitional solvent to polybed 812, then performing post sectioning immunolabeling. The tissue has "holes" in it when viewed at the TEM. They are not holes in the section, but appear to be voids in the tissue. We have tried leaving out the propylene oxide, we have osmicated, and have looked at osmotic strength of the fixative (and many variations of the above, to no avail). Can anyone offer advice? Randy Nessler Univ. of Iowa
} -----Original Message----- } From: Nancy Zjaba [mailto:nzjaba-at-alfalight.com] } Sent: Thursday, September 06, 2001 11:18 AM } To: 'microscopy-at-msa.microscopy.com' } Subject: SEM: Tilt correction factor } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hello to the list. } } It's been a while since I've been on the list, and since I've } been working } as a microscopist. } } If I image a sample in the SEM with the sample tilted, then } want to make } measurements from the images, what is the equation to use to } correct for the } tilt? I know it involves the cosine of the tilt angle, but } beyond that, I'm } drawing a blank. } } Thanks for your assistance. } } Nancy Zjaba, Technical Staff } Alfalight Inc. } 1832 Wright Street } Madison, WI 53704 } (608) 240-4875 } nzjaba-at-alfalight.com } } } }
Recently, a collegue was having problems with a white crystaline substance covering his samples of ear tissue after they were removed from the critical point dryer. It was believed to be from a contaminant in the ethanol and it was recommended to switch to acetone dehydration as it was less likely to have this problem. I haven't done SEM in years, so I haven't been paying attention to anything like this on this listserve. I was wondering if other people experiencing this problem. If you are using acetone, have you ever seen this?
Depends upon the geometry of the sample: try making the calculations for a sphere at a 45 degree angle.
My guess is that the measurements would be the same as if the sphere were at 0 degrees or 90 degrees.
For purely planar sample the equations would be valid.
regards,
Earl
----- Original Message ----- } From: "Nancy Zjaba" {nzjaba-at-alfalight.com} To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 06, 2001 9:18 AM
Karen,
One possible source for this contaminant is from the use of drying agents such as molecular sieves or other crystals. Unless one is extremely careful, the fine crystals get suspended in the liquid phase and dry down onto the specimen. We no longer use drying agents in our ethanol but use absolute ethanol directly from small (400-500 ml) bottles. If the bottles have been opened for awhile, then we use the alcohol to prepare a dehydration series since it is no longer anhydrous.
You CAN still use molecular sieves, etc. but we prepare them one month before use and take care not to jostle the bottles to any extent.
Another source for the crystalline material may be from prior contamination of the chamber by someone who had a dirty sample or contaminated ethanol.
JB
} Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
The address for your LOMO dealer is: Microscopy Labs Box 338 Red Bank, NJ 07701 732 747 6228 fax 732 758 9142 ----- Original Message ----- } From: "Bradley Starcevich" {microscopist-at-excite.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 06, 2001 10:17 AM
Once again, my plea for help to you friendly and knowlegeable folks on this listserve produced twelve hints, warnings to protect my fingers and encouragement to keep smiling through equipment hassles. I have compiled them into a lengthy word document and would be happy to send it to anyone requesting it. The following were the most important bits of advice I received from all of you and from a discussion with one of JEOL's excellent service techs working out of Pittsburgh a) the camera mechanics are pretty much the same (100, 1200, 2010 even) b) if an error message indicating the "no film loaded" appears on the computer screen, remove the fuse which turns the mechanism's motor off (preventing the motor from burning out and your fingers should you elect to remove film + plate) c) examine plates--this one was "bowed" and showed a wear pattern d) don't load the film box to capacity e) dispose of damaged plates, even those with minor bends f) examine the teflon rollers on the film boxes, replace if they do not rotate freely
Thanks again to: Sergey Ryazantsev, UCLA; Robin Cross, Rhodes University, South Africa; Keith Moulding, Hong Kong University of Science and Technology, Hong Kong; Deb Stenzel Queensland Univ. of Tech, Brisbane, Australia: . John Hunt, Cornell: John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI); Malcolm Haswell, UK; Richard Beanland, Structural Analysis Lab,Caswell Technology; Sara Miller, Duke University; Wilbur C. Bigelow, University of Michigan; Marie E. Cantino, University of Connecticut; Vitaly Feingold Scientific Instruments and Applications.
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Using clean ethanol (does not need to be "dried" for CPD) would be a lot easier than replacing seals - if the valves in your CPD don't like acetone. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, September 07, 2001 7:47 AM, Karen Pawlowski [SMTP:kpawlow-at-swbell.net] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D.
I would first like to thank the list members for the responses I have received. Thanks are also due to software support at Synoptics for a prompt and informative reply to my e-mail. I provide here a brief description of how the problem was resolved finally, in the hope that the info may be useful to somebody someday, just as I have learned a lot from this list.
On checking the binary file from the SGI computer in a hex viewer, I confirmed that the bytes were not in the format required by Semper for a Fortran unformatted file. Also confirmed that this was not due to file transfer by explicitly setting the transfer mode in ftp to binary. The e-mail from Synoptics informed that the Fortran unformatted format was not intended to be portable across operating systems and different hardware and that the input / output commands in Semper supersede the read / write commands. I discovered that the format is not even portable from the DOS version to the Windows version of Semper. The main suggested solution was to convert the image to one of the standard binary image formats such as tiff that can be read into Semper.
Two solutions are possible: (i) the tv1 program in EMS can be used to display the EMS image and the screen can be exported to Semper format. This export file has the extension eix and is an ASCII file with the image pixel data as integers. When read into Semper it is available as a byte class image. Problem with this approach is that the pixel data floating point values are converted to 8-bit values and I am not sure if this affects further quantification. Also, it requires selecting the area to be saved and I tend to end up with odd sized images which included bits of the screen background requiring further cropping with Semper.
(ii) To change the source code of se1 such that all unformatted Fortran write statements are replaced with formatted write with the format strings as per Semper file format specifications. On compiling and running, this results in an ASCII file which can be read into Semper as a floating point image, that is, without any change in the pixel values. This is the solution I have adopted. The resulting file size is larger by about 3.5 times but this is not a limitation for me. I am still left with the minor problem that the title of image is getting written as a string of blank characters. My Fortran is a bit rusty and I hope to solve this soon.
Best Wishes to all.
-----Original Message----- } From: Divakar [SMTP:divakar-at-igcar.ernet.in] Sent: Tuesday, September 04, 2001 10:33 AM To: 'Microscopy-at-sparc5.microscopy.com'
Hi Alan, Go get em! I have a Camera Lucida for a Leitz Ortholux that I have sitting next to me in my office. The collar that I have fits over the narrow diameter eyepiece tube on my scope and sets my Camera Lucida at the exit pupil of the ocular. This permits very close concordance of the two images transmitted by the prism. Of course, my camera Lucida is an old thing with a 4x6" mirror hanging off to the right. My drawing surface must be inclined, because my oculars are inclined on my binocular headed microscope. I had to construct the inclined table, but it works anyway. Here's a project. Take a piece of paper and mark it off in half centimeters (i.e., every 5mm, Ho! Ho!). Using a high mag objective look at something small like a diatom. Using the graduations on the fine focus knob change focus by one graduation and move the paper one 5mm increment drawing the outline of the in-focus diatom at each point. You will quickly see how to limit your lines so they don't overlap. Try to predict what the result will be after you draw several outlines. The key to getting everything right is to balance the illumination of the paper with that of the scope, but you probably noticed that already. Send me a photo by direct eMail. The listserver doesn't convey attachments.
You are welcome to ask questions any time or to discuss, What kind of scope(S) do you have? Illuminators? Use high intensity on the paper!
Regards,
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail (Work): fmonson-at-wcupa.edu eMail (Home): monson_fc-at-msn.com Please call before visiting
} ---------- } From: Alan Davis } Sent: Wednesday, September 5, 2001 7:16 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Camera Lucida drawing tube (Zeiss) questions } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have gotten ahold of a Zeiss camera lucida drawing tube. It has a } mirror at an angle that allows one to draw underneath the inclined eye } tube. Works for a Zeiss Standard 16. I am getting the hang of it a bit. } However, I have at least one question: } } What is the collar for (referred to as "ring clamp" below)? It is } somewhat eccentric, but I cannot imagine any purpose for it. The device } doesn't fit over it, but nearly does. It *is* useful for attaching it to } a wider eyepiece tube. } } This plastic collar fits snugly inside the eyepiece tube of an Olympus } SZH; this is good. Also, it fits over the end of the standard 16 eye } tube, with a barrier that doesn't allow it to slip down over the tube. i } have some documentation for a similar drawing tube, but there is a slight } difference in the picture (there is a knurled knob at the point where the } mirror/prism fits over the eyepiece. Mine doesn't have that, it fits } snug). Here is the relevant section from the documentation for the } pictured device: } } 1. Focus microscope. } } 2. Remove eyepiece and slip ring clamp on to tube. Replace eyepiece } and fix with ring clamp. [I referred to the "ring clamp" as a } collar.] } } 3. Clamp drawing prism on the eyepiece. [I assume this refers to the } knurled } screw that isn't present on my device---mine doesn't need to be } clamped on.] } } } } } From reading the description, though, I don't know what it is for. Is it } a spacer to move the eyepiece up higher? Which Zeiss is it really } designed for? } } Also, I would greatly appreciate any tips. The tips I found in MICSCAPE } were helpful. Can I initiate some correspondence with someone who has } some experience with one of these fantastic devices? } } Alan Davis } Marianas High School } } -- } adavis-at-saipan.com 1-670-235-6580 } Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI } } I have steadily endeavored to keep my mind free, so as to give up any } hypothesis, however much beloved -- and I cannot resist forming one } on every subject -- as soon as facts are shown to be opposed to it. } -- Charles Darwin (1809-1882) } } } } } }
According to Prof Bigelow, one clears such a jam with a wiggle! But from the Eastern Hemisphere and Dr. Moulding we hear that such jams are freed by a waggle. Now what needs to be established is whether one should wiggle or waggle, or whether one must learn to do both, AND whether TEM's behave differently toward wiggling and waggling in different places (We all know JEOL is 100V no matter where you find it.). Having been raised in North America, I have been exposed to what has always been described as typical wiggling, and I would hope that if I ever saw someone waggle, I would know that waggling was being done. One might be terribly embarrassed if one were to characterize a wiggle as a waggle. For example, is there wiggling or waggling in typical down-under dancing? If one stands in Hong Kong ogling the girls as they pass by, is one traumatized by the wiggle or the waggle? We know that one can wiggle a nose, but conversely one can only wag one's tail - in North America. But then, we all have seen buns wiggle. Perhaps the relationship is something like this. A waggle could be a small wag, and since we know that Prof. Bigelow was not suggesting that one should wag the plate to free it, rather he suggested that we just give it a little wiggle. In this way one could assert no difference between a wiggle and a waggle, both being, similarly, small wags. One would not say, "Wag the plate a little." But one could "Waggle it!" or just "wag it a little!" This would all be so much more clear if dogs in North America were known to "wig their tails" when they are happy. Perhaps what we have is an hemispheric etymological inversion such that in the West, wiggle is to wag, where as, in the East, waggle is to wig! We here in the Western Hemisphere have learned to watch worms wiggle, but what do they do elsewhere, I wonder? Might a waggle merely be out of phase with a wiggle? Sine! Cosine! Do these differences occur, because of the differences between 50 and 60 Hz? Perhaps there might be NSF or WHO money for such a question.
Oh well, ruminating like a mad sow, he demurred, "No rumen in the old sow at all."
If anyone can clear any of this up, I certainly would like to know!
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
While I have usually come to a comfortable resolution with respect to this kind of safety issue, this one has proved elusive. So.....
My normal procedure is to unwrap and clean the ampoule as I would my best glassware. That is, I treat it to a "biochemistry lab" cleaning, finishing with 3X rinses in sgdd(single glass-distilled deionized) HOH. Between the last two waters AND WEARING FRESH POWDER-FREE GLOVES, I score the ampoule three times with an ampoule file while holding it on a folded paper towel in the bottom of a shallow heavy-glass bowl. I then transfer the ampoule to an appropriate 100ml bottle (I use a serum bottle, because it is constructed of thick glass, shows imperfections readily (especially around the bottom), and can withstand the breaking of the ampoule). The scored ampoule breaks more readily than an unscored one, and three scores (~1cm apart) provide sufficient weakness for breakage to occur with relative ease. To further facilitate breakage, I add only some of the total amount of solvent HOH (for me, 25ml) sufficient to keep the ampoule from floating while providing enough for initial immersion of the crystals. Finally, I rest the serum bottle on the same folded paper towel in the same thick glass bowl to dissipate the shock of the breaking regimen. NOTE: Since it is at this part of the process at which failure has occurred (twice in 25 years the bottom of the bottle has separated from the barrel), I am still working in the hood, but with a substantial amount of 95% EtOH at hand to render the OsO4 harmless if I have an accident.
If the above smacks of science, please forgive the allusion. The method and approach are the result of what has evolved over time through trial and error and the application of common sense to a continuing, natty problem.
I have always hoped that someone would supply a FINAL solution so we can all be more comfortable when "Stock solution" time arrives.
Regards,
Fred Monson
} ---------- } From: Patton, David } Sent: Friday, September 7, 2001 5:30 AM } To: Monson, Frederick C. } Cc: 'Microscopy Listserver' } Subject: Question about preparing osmium tetroxide } } While we are on the topic of making up fixatives I have a } supplementary question. } } How do you open glass osmium tetroxide ampoules? } } I put mine in a 100ml glass bottle and then break it with a } glass rod. This is nerve-wracking, tedious (they do not } break readily) and not as safe a procedure as I would like. } } The ampoules used to have a weakened neck which could be } sawn off but they do not come in this form from our } supplier any more. } } Is there an easier way? } } Dave } } } On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." } {fmonson-at-wcupa.edu} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a } bottle } } with a Teflon lined screw cap. The bottle is then held in the } refrigerator } } in a small plastic jar (available from all lab distributors - mine from } } Fisher or S/P) with a screw cap . In this way, the plastic becomes } } osmicated NOT the refrigerator. I have done this for years with no } untoward } } results. } } } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) } glass or } } plastic. Osmication is done in glass vials with Teflon lined screw caps } } using a minimum of OsO4 to do the job. } } } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml } dd } } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with } a } } Teflon lined cap. I then make up an appropriate amount of fixative } using } } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is } kept } } in a plastic jar in the refrigerator. } } } } I find that double containers are safe and reduce contamination } } considerably. } } } } Hope this helps. } } } } Fred Monson } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging(CASI) } } West Chester University of Pennsylvania } } Schmucker Science Center II } } South Church Street } } West Chester, PA, 19383 } } eMail: fmonson-at-wcupa.edu } } http://darwin.wcupa.edu/casi/ } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
Depends also from the measurement direction. The formula is right in the tilt direction, but the real tilt angle of a straight line on flat sample varies from nominal tilt angle in tilt direction to zero in the perpendicular direction. In the other directions, the formula is right too, but you must estimate the real tilt angle in the measurement direction, in respect to the tilt direction.
And of course, don't forget than in a SEM the perspective is not "conique" (I don't know the right terms in english) but "cavalière" . A tilted square looks like a rectangle and not a trapezoid. Try with a TEM grid with square holes, and try too with a grid with hexagonal holes. And rotate it ... You may feel dizzy, with high tilt angles. Our brain is not build for such a perspective ! But it can give nice pictures.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Depends upon the geometry of the sample: try making the calculations for a } sphere at a 45 degree angle. } } My guess is that the measurements would be the same as if the sphere were at } 0 degrees or 90 degrees. } } For purely planar sample the equations would be valid. } } regards, } } Earl } } ----- Original Message ----- } } From: "Nancy Zjaba" {nzjaba-at-alfalight.com} } To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, September 06, 2001 9:18 AM } Subject: SEM: Tilt correction factor } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello to the list. } } } } It's been a while since I've been on the list, and since I've been working } } as a microscopist. } } } } If I image a sample in the SEM with the sample tilted, then want to make } } measurements from the images, what is the equation to use to correct for } the } } tilt? I know it involves the cosine of the tilt angle, but beyond that, } I'm } } drawing a blank. } } } } Thanks for your assistance. } } } } Nancy Zjaba, Technical Staff } } Alfalight Inc. } } 1832 Wright Street } } Madison, WI 53704 } } (608) 240-4875 } } nzjaba-at-alfalight.com } } } } } } } }
The method we use, which I teach to all the students, is to wrap the vial with a short length of paper towel (usually about four or five turns around the bottle), grasp the vial at both ends, and gently exert pressure with the thumbs. Then carefully unwrap the package and throw both vial halves in warm distilled water. I've never had any problem, either with crystals falling out in the paper or with glass poking through the paper towels. I was also taught the whack method, and found that my aim was not adequate to the job.
We use the same wrapping method with glutaraldehyde ampoules, but instruct the students to be sure to keep them upright.
Marie } } While we are on the topic of making up fixatives I have a } supplementary question. } } How do you open glass osmium tetroxide ampoules?
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Regarding tilt correction, I enclose this from an earlier reply: it is a warning against the use of the tlit correction control on the SEM.
*** one thing is that [certain] morphology is generally quite low-profile and vague under SEM, _unless_ you tilt the specimen: I generally tilt at 54^ = ARC COS (1 / SQRT(3)), the "magic" angle which ensures that x, y, and z come out isometric, with the axes at 120^ to each other: if there is any orientation (say in an injection moulding or drawn specimen, then while the specimen is still flat-on, make sure that x and y are both at 45^ to the tilt axis. Then you will get isometric projection of all three axes. But NEVER use the tilt-correction control, or you will get distortion: for example, it makes spheres (which should appear circular at any angle) come out as ellipsoids.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I have a diamond pen or scribe (bought mine from EMS) that I normally use for indelibly etching numbers on glass slides. I use it to carefully scratch a circular line completely around the narrowmost point of the ampoule. I usually go around several times to ensure a good scratch. I rinse the vial off with 95% ethanol and place inside a sturdy glass bottle (I use a Schott-brand glass bottle with orange plastic cap - available from major supply houses like fischer). I then shake the bottle vigorously (inside the fume hood, while silently praying it doesn't break!) It generally breaks within a few good snaps of the wrist. I add 50 mls of water and then store this bottle inside a plastic jar in my fume hood. The inside top of the plastic cap of the Schott bottle and the inside of the plastic jar go black with time and I periodically (yearly) replace them. I would never trust putting this stuff in my refrigerator. It leaks out the first bottle - why would one think it doesn't then leak out the second? I use it up usually within 3-4 months and never had a problem with room temperature storage over the last 20 years. If you use ampoules with pre-scored lines (easy snap open style) there is another trick to consider i used to use in the old days but it comes with a potential hazard that safety officers and lawyers advise against so I won't recommend it. Place about one cm of liquid nitrogen in a 50 ml disposable beaker in the hood. Slowly lower the ampoule into the liquid nitrogen. this causes the osmium to contract and break free from the glass. Snap the ampoule and pour the contents into water. the low temp also reduces fumes during pouring. this method avoids glass in your stock solution. the danger is that if there is a crack in your vial, the nitrogen would enter, and when warmed, potentially explode the vial. I never tried dry ice but i suspect it would work and avoid this hazard.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have done CPD for years, and never seen this problem with EtOH dehydration (or acetone). The only times I've gotten anything like this is from the specimen container used for drying, or salt precipitation when the buffer was incompletely washed off. Another possibility is the CO2. Many CO2 tanks are full of water and crud. Is there a molecular sieve filter (for the water) and a membrane filter (for the crud) on the line from the tank to the CPD?
Mind, it contaminant *could* be from the EtOH, but that's just a reason to use better EtOH, not to switch to acetone.
Phil
} Hi, } } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D.
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hello all, I was wondering what computer programs people are using for 3-D reconstruction of TEM serial sections. I am interested in peoples opinions of the software (i.e. ease of use, learning curve, cross-platform functionality, cost, etc.). Feel free to email me off list if you choose. Also vendors feel free to send me any info you might have on the products you sell.
Thanks to all,
-EK
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology II Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
see our web page at http://www.mayo.edu/mcniven_lab/
Another source that we have come across is sodium phosphate buffer crystals. If fixation is carried out with sodium phosphate buffered soluitons, and then immediately transittioned to dehydration solvent (either EtOH or Acetone) then we have seen samples coated with fine crystals. We use either serveral water rinses between fixation and dehydration or an alternative buffer (i.e. Na Cacodylate)
} } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D. } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I would be interested in hearing from people anywhere in the world who have had PERSONAL experiences with the installation of electron microscopes or other sensitive equipment within 800 meters of electrically powered light rail, trains, buses or subway systems which have created vibrations or stray field problems for these instruments. I am specifically interested in hearing from those who have had prior experiences with light rail or bus transportation units powered by electricity. I would like to hear positive (no known problems for sensitive instruments) as well as negative experiences you have had or are having.
If you are aware that one or more of these types of transportation systems is within the 800 meter range of your equipment and specific measures were taken to eliminate vibration or stray fields, I will like to know that too.
You may wish to respond to me personally. Thank you.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
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Dave, I personally would not use a supplier who does not pack Osmium in easy open vials or ampoules such as the ones which have the thin neck that can be snapped open. It is just too dangerous to try to break the vials as you described.
Often the osmium crystals will adhere tightly to the glass. If you dip the vial into liquid nitrogen while holding it by the neck with forceps, the crystals will release from the glass surface. They can then be shaken down to one end of the vial.
You can purchase silicon ampoule breakers from some of the major EM supply houses. They are flexible molded silicon with a depression the shape ofthe ampoule. Just put the ampoule in, bend the mold and the ampoule will break. Then remove the portion with the crystals and pore them into your osmium container. You end up with the two large piece of glass which should be disposed of carefully since there might be traces of osmium remaining. The osmium crystals will normally dissolve into water (or buffer) if left overnight at room temperature....in a hood.
I normally make osmium up double strength in water and it will keep for months in the refrigerator, remaining a clear light amber solution. I also recommend double bottling to help prevent escape of vapors. However, we do not store any biologicals in the same refrigerator with fixatives. Regardless of how careful you try to be, the odds are that some fixative vapors will escape.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Gray, Peter, 1975, Microtomist's Formulary and Guide, R. Krieger, Huntington, NY. (Still available for the hardy bibliophile at http://WWW.WEB4U.COM/cgi-bin/full_page?0-88275-247-2) reports FOUR fixative mixtures that are attributed directly to Champy:
My favorite, of course, is number 2!!! whose last component provides a wonderful bit of chemical history. We see how Bayer must have started. Dupont.
Regards, and I know that this doesn't help at all!
But it is all part of the art.
Fred Monson
} ---------- } From: Richard Edelmann } Reply To: edelmare-at-muohio.edu } Sent: Wednesday, September 5, 2001 4:24 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Bio. Fixation: What is Champy's Fluid? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed } by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
EMS sells an ampoule breaker , Cat# 60600 for $12. We've used them for breaking ampoules of paraformaldehyde, glutaraldehyde, acrolein and osmium within seconds. The soft silicon molds can be twisted to achieve a quick break. I usually tap the osmium tetroxide crystals to the bottom and use forceps to lift the open ampoule into my prepared 50 ml of distilled water to make our stock 2%. I have no financial interest in this company but am a satisfied customer for many years. Phone number is: (800) 523-5874. Rosemary Walsh, EMF for the Life Sciences PSU
Fred: Have you given any though to the fact that Coriolis' forces are different in the southern hemispher? Water drains from a basin in a counterclockwise motion south of the Equator and clockwise north of the Equator. Maybe wiggles and waggles are affected thesame way between the eastern and western hemispheres.
Regards,
Sam Purdy
Technical Center National Steel Corp. 1745 Fritz Dr. Trenton MI Voice:734-676-2682 FAX: 734-676-2682 spurdy-at-nationalsteel.com
Use a triangular metal file to score around the neck, then use the usual precautions (protect fingers, open in hood , etc.) to pop open the vial. The EMS brand I use breaks open easily by wrapping paper towel/ gauze around top and gentle application of pressure.
Becky Garrison I am not an employee of any EM supplier.
-----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Friday, September 07, 2001 5:31 AM To: Monson, Frederick C. Cc: 'Microscopy Listserver'
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
David Patton wrote: ======================================================== While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way? ========================================================== I am a bit surprised to hear that you are getting osmium tetroxide that is not in prescored ampoules. So far as I know, all of the major suppliers of EM chemicals, like SPI, have long since supplied their ampoules in the pre- scored form. The only time I have seen in recent years ampoules not in pre- scored form are those that are destined for other markets and applications.
Also, SPI, as well as the other major suppliers have long offered ampoule neck breakers, such as are described on URL http://www.2spi.com/catalog/tools/smtol21.html These are meant to be used one time and discarded. They are inexpensive and seem to provide an extra element of safety.
Using only prescored ampoules and an ampoule neck breaker might be easier on the nerves.....
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Below are the two answers to your question, since I am not sure whether you have obj. aperture inserted in the beam, or you don't. Both assuming that the field of view that you mentioning is the image of the sample in the holder, rather than an illumination circle.
1. If obj. apertures holder is inserted in the beam. Objective aperture should not be used in LM range. In Philips TEMs starting with EM400 series , the main optical configuration difference between LM and M modes is very distinct and as followed: in LM mode, the objective lens current is very low (few hundreds milliAmps, depending on HT set), and is fixed- hence magnification of obj. lens is also low (between 1 and 10, closer to 1- just a guess). Focusing in LM mode is done by diffraction lens. Almost all magnification is accomplished by lenses below the objective lens. Obj. aperture is located between the specimen and that lens system, much closer to the specimen, than to the first magnifying (diffraction) lens, and is blocking most of the image if inserted in the beam.
In M mode, the obj. lens current is much higher (several Amps, depending on the particular TEM model and the HT set), making magnification of the obj. lens in M mode much higher (say, 50 or so). You can use obj. aperture now, because of the significant part of the magnification happens very close to the obj. aperture, plus much smaller area of the specimen is forming first (objective lens) image. Focusing in M mode is done by the obj. lens. Diffraction lens current in M mode is fixed and depends on the HT and magnification set.
This two modes are "toggled" when TEM magnification is switched LM to M and vice versa, in a single magnification step.
2. If obj. apertures holder is withdrawn from the beam. When the condenser aperture is out of the beam, field of view in the lower LM range increases. Nevertheless, obj. aperture holder shall not be visible when withdrawn from the beam (control lever to the right). If obj. apertures holder is still visible, then either the column alignment, or the obj. aperture mechanical alignment (or both...) are incorrect.
If you are referring to illumination circle with the specimen holder withdrawn- I wouldn't care much about obj. aperture holder visible on the periphery of the illumination circle at the lowest LM steps, as long as it is not blocking the sample image at the corresponding extreme of the sample travel, and as long as everything else looks normal.
If you need to align the "out" position of any aperture holder on this TEM, turn little Allen set screw located on the right side of the appropriate aperture holder mechanism housing.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: "marienti-at-tiscalinet.it"-at-sparc5.microscopy.com {"marienti-at-tiscalinet.it"-at-sparc5.microscopy.com} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
How would you like to escape the northern winter this year?
Yes? Then join us for the 17th Australian Conference on Electron Microscopy, to be held in Adelaide, South Australia, from February 4 - 8, 2002. Encompassing the areas of Electron, Scanned Probe and Confocal Microscopies, this important meeting will provide an opportunity to explore the many applications of these techniques, to keep abreast of the latest developments and to meet and mix with many of Australia's and the world's eminent leaders in the field. This meeting was last held in Adelaide in 1988. A lot has changed in this city in the past 14 years. Adelaide has emerged as a vital city which can offer the visitor a range of exciting opportunities. The local wine and food have attained international recognition. Tourists are welcomed and well catered for. The conference will be held in the brand new Adelaide Convention Centre which is conveniently located in the CBD, close to accommodation and other facilities. International visitors should take some extra time either before or after the conference to explore Adelaide and South Australia. Tours can be arranged to take you to the sights of Adelaide, the nearby picturesque Adelaide Hills, and the wine growing regions of McLaren Vale, the Barossa Valley and Clare Valley. Each of these areas has its own unique history, heritage sites and gourmet delights! Adelaide's clean white beaches are within easy reach of the city. Experience the wildlife, flora and landscape of Kangaroo Island. Adelaide is also the perfect stepping-off point to locations such as the beautiful Flinders Ranges, where centuries-old Aboriginal art and ancient geological formations can be seen, Uluru, the opal mining town of Coober Pedy, and the Outback. The current exchange rates ensure that travel in Australia has never been more economical for the overseas tourist. A series of pre-conference workshops will be held to allow participants to gain hands-on experience in the latest techniques and applications. Many of the keynote speakers will participate as presenters, providing a wonderful opportunity for interaction with these noted scientists. Be sure to book early, as places in some of the workshops is limited. During the week, delegates will have the chance to mingle and relax after hours in several of Adelaide's noted heritage locations. A range of accommodation choices, from five-star to budget, ensures choices suitable for all. With the deadline for abstract submission fast approaching, (October 6), start making plans now to join us in February. Abstracts should be submitted electronically, and the registration form can be downloaded from the website. Full details can be obtained at http://www.adelaide.edu.au/CEMMSA/acem17 Keep an eye on the website for regular updates. We look forward to seeing you at ACEM17.
ACEM17 Organising Committee
CEMMSA Centre for Electron Microscopy and Microstructure Analysis Adelaide University Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
At 10:30 7/09/01 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
we 1. thouroughly clean off any label and label glue from the vial
2. put a scratch right around the vial
3. THEN it becomes easier to smash in the bottle, as you do it.
AND if you want, you can promote a crack around the line of the scratch by touching a redhot glass rod to the scratch.
AND if you want, you can distribute the OsO4 in a thinner layer around the wall of the vial by warming the (unscratched) vial in hot water, then rolling the vial as the OsO4 cools.
This accelerates solution.......
AND if your OsO4 starts to go dark, you can refresh it almost completely by adding hydrogen peroxide dropwise and thus titrate it to a clear solution again. Oten you can restore it to straw colour, but we will use even a pale purple solution as it will have enough tetroxide in it to work.
Upon reading subsequent posts about breaking OsO4 vials, I realized I must amend my suggestion of wrapping the neck with paper towels and gently applying pressure to break at the neck. This method works fine on PRE-SCORED vials, but almost certainly is unsafe or ineffective when used on unscored vials. I wasn't aware that unscored vials were still being sold.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
I have an Hitachi H7100FA TEM on the second floor of a building built almost directly over an underground electric railway. When I moved in, bringing the EM with me, there was the choice of various locations. I spent very little time standing on a wooden floor feeling it shake under me. I spent considerably more time sitting on a concrete floor with a saucer of water watching for vibrations (we had no money to pay experts). Apart from having someone ask me if I needed medical help, I was left in peace with my saucer and am now quite happy with the chosen location. I don't work at REALLY high mags, but the resolution test was accomplished without any vibrations and I've coexisted happily with the trains for 3 years. Before that I was on the 2nd floor of a concrete building next to a busy road and it was dreadful. The building construction and location within it was the important aspect for me.
Diana
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Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
We are currently running an Oxford ISIS EDS system and an Oxford MonoCL cathodoluminescence system from the same computer and on the same SEM. We want to split the Cathodoluminescence system off onto another column so we need another ISIS board enclosure (box).
We would be pleased to hear from anyone upgrading their system who would have an ISIS box that is spare. (The box that houses the ISIS boards, not the PC.) We do not specially need the ISIS boards.
We will pay a reasonable amount (say US$5,000) plus shipping for an ISIS box in working order.
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
I'd like to prepare catalyst particles for TEM viewing by ultramicrotomy. The problem is that I want to study the location and nature of carbon in these particles and I am not certain how to distinguish the carbon from the epoxy. We currently use Spurr's resin, but is there an epoxy I can use that is not carbon based ?
TIA Willem Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 960 - 4211/2772 Fax : +27 +16 960 - 2826 E-mail : willem.erasmus-at-sasol.com PO Box 1, Sasolburg, 1947, Republic of South Africa
[All views expressed are my own and not necessarily that of my employer.]
Could be interesting for you... ----- Original Message ----- } From: "Steven Dessein" {steven.dessein-at-BIO.KULEUVEN.AC.BE} Dear Taxacomers,
The Laboratory of Plant Systematics (K.U.Leuven, Belgium) today released Carnoy. Carnoy is an easy-to-use image analysis program, designed to carry out measurements on digital images and especially on micrographs obtained from light or electron microscopes.
Carnoy features a unique one-click calibration option: click on the scale bar, enter its length and you are ready to carry out real-world measurements on your micrographs. Carnoy measures length and surface area of regular and irregular objects. It also performs automated particle analysis (counting and measuring). The program can handle BMP, PICT, Photoshop, JPEG, GIF, PNG, SGI, TGA, TIFF and QuickTime image files and can convert between these file types. Measurements are stored in a list that smartly manages all data. Every measurement can be annotated. Measurements can be exported to a tab-delimited file, which can be read by any text editor, word processor, database or spreadsheet, for further analysis. The program's clean and easy-to-use interface enables you to be productive within minutes.
Carnoy can be used in the following fields: taxonomy, morphology, ecology, physiology, molecular cell biology, agricultural sciences, paleontology, archaeology, among others. Carnoy requires a Macintosh running Mac OS X or a PC with at least 64 MB RAM and Windows 95 / 98 / ME / NT / 2000 / XP. A resolution of 1024 x 768 pixels (or higher) is recommended.
Carnoy is $15 shareware (if you cannot afford this, please, contact peter.schols-at-bio.kuleuven.ac.be). It is available for download on the Carnoy website. http://www.carnoy.org
Kind regards,
P. Schols, S. Dessein & E. Smets
Steven Dessein Laboratory of Plant Systematics Institute of Botany and Microbiology Kardinaal Mercierlaan 92 B-3001 Leuven Belgium
I'm not sure if our Osmium vials are prescored or not, but we've never had any problems or the nerve wracking experiences that you have mentioned. We score around the vial a couple of times with a diamond pen(made especially for marking on glass). The side where we want the break is scored a couple times more. In the fume hood we wrap the vial in a paper towel and snap it facing toward the back of the hood. Hope this helps.
Paula Moore Wake Forest University School of Medicine Pathology/EM Lab pmoore-at-wfubmc.edu
} } How do you open glass osmium tetroxide ampoules? } } I put mine in a 100ml glass bottle and then break it with a glass rod. } This is nerve-wracking, tedious (they do not break readily) and not as safe } a procedure as I would like. } } The ampoules used to have a weakened neck which could be sawn off but they } do not come in this form from our supplier any more. } } Is there an easier way? } ========================================================== } } } Using only prescored ampoules and an ampoule neck breaker might be easier on } the nerves..... } } Chuck } } ==================================================
Amray 1610 SEM with PGT EDS free to a good home---interested party pays all shipping charges. Please send queries off-list. Thanks!
Melissa Troost Electron Probe Instrumentation Center Northwestern University Evanston, Illinois, USA 847-491-3439 m-troost-at-northwestern.edu
________________________________ Legal Notice: This message is confidential is intended for the stated addressee(s) only. Access to this e-mail by anyone else is unauthorized. If you are not the intended recipient, please inform the sender immediately.
Does anyone have or know of a source of SEM photomicrographs of bacteria, mold and yeast. I am asking on behalf of a colleague who will pay a royalty for the use of the photos.
Mark Germani, Ph.D. MicroMaterials Research, Inc. 136 Shore Drive, #200 Burr Ridge, IL 60521
Anyone out there knows how to run up an X-ray film (exposed as well as unexposed) for TEM?????
Dear Neelima, What do you mean by "run up"? We are lucky to have our LoDose and MRF32 x-ray films cut to the appropriate size--6.5 x 9 cm--and we treat them like other EM film EXCEPT, we load and develop them in total darkness, and we use x-ray film developer in the tank instead of D-19. Our darkroom is in the same room as the scope, so transport can occur in total darkness. If this is not the case for your facility, and you have to run the film up to your darkroom, get a film transport bag from a photo supply store, put the film in its original box, put the box in the bag, and carry it to your darkroom. The bags are designed to be light tight. Good luck, and if you mean something else by "run up", let the list know, and I'll try to give you a better answer. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Karen: Stick to absolute ethyl alcohol (Et0H). Never use acetone with any critical point drying apparatus that have glass viewing windows held in place with a epoxy resin that would be attacked by acetone. The white crystals may very well be buffer salts. You can start dehydrating with a larger ratio of water to ethyl alcohol and let stand for a longer time to remove buffer salts while working up to absolute strength ethanol. For some samples such as tissue culture monolayers the critical point dryers can be avoided altogether by proceeding from the absolute ethyl alcohol to 1,1,1,3,3,3-hexamethyldisilazane to air dry. I would recommend the 99.9% pure solution.
Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences State University of New York-Binghamton Binghamton, NY 13902-6000
} ---Original Message------------------ } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D. } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I got a problem when preparing TEM foils of Al-alloy by means of electro-polishing. A dark film produced on the specimen surface when it was been polishing. I think, the polishing condition is right and listed below:
Solution, 25 % Nitric acid plus 75 % Mechenol Temperature, about -30 degree C Voltage, 15-20V
I would be pleased to hear from anyone who has an idea to sort out this problem.
Dear Randy, Drosophila brains are very hard to fix. The neurons are very delicate and lose their structure when not well preserved. What age are the bugs? You might try some adding some acrolein (we used 1% acrolein and 2.5% glutaraldehyde in 0.1 M cacodylate buffer, pH 7.1) and leave out the paraformaldehyde. See if you still have immuno-reactivity You don't need to use propylene oxide. You can use 100% EtOH instead. It is miscible with resins, as long as it is totally dry. Also, don't store your specimens in buffer, but process straight through. I do microwaving these days, so time is never an issue anymore. Good luck!
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
04:18 PM 09/06/2001 -0500, you wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In conjunction with the Pacific Northwest Microscopy Society, The University of Washington Core for Communication Research, Center on Human Development and Disability and the W. M. Keck Center for Advanced Studies in Neural Signaling present:
Microscopy and Digital Imaging: Advances & Applications
September 27, 2001 12:30 - 5 p.m. Location: CD 150
12:30 "Multiphoton and Multispectral Imaging in Vertebrates" Mary Dickinson, Caltech
1:15 "Some New Approaches to Intracellular Imaging" Bob Fern, University of Washington
2:00 FRET imaging with the confocal microscope Kolja Wawrowsky, Leica
2:45 Break
3:15 "In Vivo Microscopy of Murine Brain" Liz Yoder, UCSD
4:00 "Deconvolution Methods in Wide-field and Confocal Microscopy" Paul Goodwin, Applied Precision Inc
5:00 Reception
The Veteran's Administration Hospital and Medical Center Morphological Analysis Core Facility will have an open house and talks on September 28, 2001.
Speakers sponsored by Applied Precision, BioRad, Leica, and Zeiss The reception is sponsored by Bartels and Stout, Inc.
Contact Paulette Brunner pbrunner-at-u.washington.edu or Glen MacDonald glenmac-at-u.washington.edu for information.
Regards, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax **************************************************************************
C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sbcook-at-bcpl.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, September 9, 2001 at 22:52:34 ---------------------------------------------------------------------------
Email: sbcook-at-bcpl.net Name: Katelyn Cook
Organization: St. Michael the Archangel
Education: 6-8th Grade Middle School
Location: Baltimore, MD
Question: I am planning to do a science fair project about germs and the importance of hand washing. I want to collect germs from common public places such as doorknobs, handrails, telephones, sink faucets, etc. What is the best way to collect the germs? Will I need to grow the germs in a petri dish after I collect them? Will I be able to identify different types of germs using my microscope?
I have seen cellular voids from poor fixation as well as from genetic mutations. The biggest impediment I have when fixing drosophila brains is getting the fixative to the tissue. I recommend cutting the neck as well as the proboscis and then using gentle centrifugation (microcentrifuge ~2000rpm for one minute) to get the heads to sink. Due to air in the brain cavity this last step does not always work. If you use 2% glutaraldehyde and osmium post fixation do you get good ultrastructure? In a project I did some time ago I had the best results using 0.07 M sodium cacodylate buffer at pH 7.5. That gives an osmolarity of about 187mOsm. However, I routinely use 0.1 M sodium cacodylate and find that adequate.
At 04:18 PM 9/6/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Liststers; } } I'm working with SEM of pre-colonial potteries from Pantanal at the = } Brazilian West border with Bolivia. In some samples we've found shell = } and sponge spicule in the bulk of potteries. We would appreciate any = } article or book with micrographies of similar materials in pottery ? = } Also, if someone could provide me references on clay with sponge = } spicule, would be very helpful. } } Ubirajara Pereira Rodrigues-Filho Instituto de Química de São Carlos Universidade de São Paulo São Carlos, Brazil
Cameca MBX Microprobe is available at this time. 4 wavelength spectrometers (includes PC2 crystal for Lt ele. spec.) Kevex Sigma EDS New GW Backscattered system PC Based operating system Diff pump vacuum system Cathodoluminescence system (OEM) System is about 17 years old and in good condition (at last service call). Has been turned off for 1 year.
Please send inquires to E-MAC Walter Protheroe corvos-at-aol.com
You may want to try the following website at Utah State University:
http://bioweb.usu.edu/emlab/Galleries.html
They have many great SEM images. A contact person there to talk to would be:
Bill McManus TEL: (435) 797-1920
I hope this helps.
Best regards-
David
Mark Germani wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone have or know of a source of SEM photomicrographs of } bacteria, mold and yeast. I am asking on behalf of a colleague who will } pay a royalty for the use of the photos. } } Mark Germani, Ph.D. } MicroMaterials Research, Inc. } 136 Shore Drive, #200 } Burr Ridge, IL 60521 } } (630) 325-8170 } (630) 325-8178 fax } } mgermani-at-micromaterialsresearch.com
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
Dear Randy, Drosophila brains are very hard to fix. The neurons are very delicate and lose their structure when not well preserved. What age are the bugs? You might try some adding some acrolein (we used 1% acrolein and 2.5% glutaraldehyde in 0.1 M cacodylate buffer, pH 7.1) and leave out the paraformaldehyde. See if you still have immuno-reactivity You don't need to use propylene oxide. You can use 100% EtOH instead. It is miscible with resins, as long as it is totally dry. Also, don't store your specimens in buffer, but process straight through. I do microwaving these days, so time is never an issue anymore. Good luck!
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
04:18 PM 09/06/2001 -0500, you wrote:
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Hello everyone, I have a researcher here that is having some difficulty with getting good histology at the EM level. She is fixing Drosophlia brains with 4%para/0.5% glut in phosphate buffer pH7.2, no osmium post fix, ethanol dehaydration, propylene oxide as a transitional solvent to polybed 812, then performing post sectioning immunolabeling. The tissue has "holes" in it when viewed at the TEM. They are not holes in the section, but appear to be voids in the tissue. We have tried leaving out the propylene oxide, we have osmicated, and have looked at osmotic strength of the fixative (and many variations of the above, to no avail). Can anyone offer advice? Randy Nessler Univ. of Iowa
Yan, Assuming (sorry for that) that the polishing conditions are correct, and you are producing well dished samples with thin area and the only problem is sludge on the surface, try two things before changing any other conditions and try them one at a time so as not to confuse the issue. 1: Try increasing the jet speed. If this doesn't work. 2: Try increasing the temperature about 5 degs. It would be helpful to know what type of jet thinner you are using and exactly what the alloy is. Good Luck! Dorrance
} ---------- } From: yan.hu } Sent: Tuesday, September 11, 2001 12:35 AM } To: microscopy-at-sparc5.microscopy.com } Subject: ask help } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } I got a problem when preparing TEM foils of Al-alloy by means of } electro-polishing. A dark film produced on the specimen surface when it } was been polishing. I think, the polishing condition is right and } listed below: } } Solution, 25 % Nitric acid plus 75 % Mechenol } Temperature, about -30 degree C } Voltage, 15-20V } } I would be pleased to hear from anyone who has an idea to sort out this } problem. } } } Regards } } Yan } ---------------------- } } yan.hu-at-brunel.ac.uk } } }
On Tue, 11 Sep 2001 05:22:19 -0500, sbcook-at-bcpl.net wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Below is the result of your feedback form (NJZFM-ultra-55). It was | submitted by (sbcook-at-bcpl.net) from | http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, | September 9, 2001 at 22:52:34 | --------------------------------------------------------------------------- | | Email: sbcook-at-bcpl.net | Name: Katelyn Cook | | Organization: St. Michael the Archangel | | Education: 6-8th Grade Middle School | | Location: Baltimore, MD | | Question: I am planning to do a science fair project about germs and | the importance of hand washing. I want to collect germs from common | public places such as doorknobs, handrails, telephones, sink faucets, | etc. What is the best way to collect the germs? Will I need to grow | the germs in a petri dish after I collect them? Will I be able to | identify different types of germs using my microscope? | | --------------------------------------------------------------------------- |
Dear Katelyn Cook - While I was teaching as a graduate teaching assistant for a general biology class, I had the opportunity to lead a group of non- science college students on an examination of the microbial world around them. What I did was to have each of them use a sterile, cotton- tipped swab, moisten it a bit with sterile water (perhaps boiled water will do just as well), swab the surface of whatever object they wanted to explore, and then swab it onto the surface of a petri plate that already had bacterial medium. They then left all the plates for me to seal and incubate at room temperature for several days (or, at most, 1 week). When they got the plates back, they then could see with the naked eye the various colonies that formed and their various colors. If a microscope is available, then it is possible to identify individual bacteria by using a "wet mount" (that is, a slide with a small drop of sterile water and a very small amount of whatever colony is chosen). Although I don't have any experience with grade school students, I believe that my approach could work. So... to answer your questions: (1) A very easy way to collect bacteria is to first get some sterile swabs. The fact that they are sterile is important because any box of ordinary (or household) swabs is likely to already have bacteria on them, and obviously that would defeat the purpose of your experiment. As a result, you'll need some sterile swabs -- quite likely, they can be bought at a pharmacy near you when you look for dressings, bandages, etc. (2) Assuming you have sterile swabs, you could conceivably boil some water, let it cool to room temperature, and let each student lightly wet a swab (or more as needed). Why boil the water? For the same reason as needing sterile swabs -- because ordinary tap water will very likely contain bacteria (even if it looks perfectly clear). One alternative is to use distilled water if boiling isn't feasible. (3) Yes, you will need a petri plate -- the plate needs to have some kind of bacterial medium capable of supporting their growth; otherwise, your swabbing exercise will be fruitless. I do not know where such plates can be obtained, though, and you may want to seek others on this listserver; finally, (4) The use of a microscope is needed only if you're interested in examining individual cells. The typical colonies that are seen on the surface of a plate tend to vary in terms of both color and the type of colony produced. That is, a colony could be white or yellow, but its 'type" could be of a sticky nature or a more "liquid-like" nature. That part could be revealing on its own, or perhaps not. If you feel that a microscope is needed, then a simple way to see individual cells is to simply add a small drop of boiled / sterile water onto a slide, use an inoculating loop or needle, and gently pick up a small amount of a colony, and spread it around within to the drop. Place a coverslip over the slide, and put it onto the stage of a microscope. Try to use the highest objective lens magnification that is possible -- if there are a 10X, 20X and 100X objectives, use the 20X as the individual cells are very small in size -- and, if it's possible, try to set the microscope under non-brightfield light conditions such as phase-contrast. Bacterial cells are, generally, transparent to light, and as a result are often very hard to see because they have very little contrast with the background light. Anyway .. I hope you have some success with your experiment for these grade-school students. It is my my belief that microscopic, living things (such as the "germs" and, more specifically, bacteria, algae, etc.) are always fascinating things for kids to see, and I hope it turns out very well. Good luck ! Nelson Conti Former graduate student at San Francisco State University (in San Francisco, CA) A Bachelor's and Master's degree in Microbiology
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Have you contacted Oxford Instruments? With the INCA doing as well as they tell me it is they must be taking in quite a few ISIS towers on upgrades (they've had one of ours). Of course if you expect a full manufacturer's warrantee it might be pricey but if you took the same 'as seen' conditions that you would accept from another user then they might deal.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } We are currently running an Oxford ISIS EDS system and an Oxford MonoCL } cathodoluminescence system from the same computer and on the same SEM. We } want to split the Cathodoluminescence system off onto another column so we } need another ISIS board enclosure (box). } } We would be pleased to hear from anyone upgrading their system who would } have an ISIS box that is spare. (The box that houses the ISIS boards, not } the PC.) We do not specially need the ISIS boards. } } We will pay a reasonable amount (say US$5,000) plus shipping for an ISIS } box in working order. } } } } } Dr. Mel Dickson, } Deputy Director, The Electron Microscope Unit, } Adjunct Associate Professor, School of Microbiology & Immunology } The University of New South Wales } UNSW SYDNEY 2052 } Australia. } Phone +612 9385 6383 Fax +612 9385 6400 } http://srv.emunit.unsw.edu.au/ }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
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EMS sells an ampoule breaker , Cat# 60600 for $12. We've used them for breaking ampoules of paraformaldehyde, glutaraldehyde, acrolein and osmium within seconds. The soft silicon molds can be twisted to achieve a quick break. I usually tap the osmium tetroxide crystals to the bottom and use forceps to lift the open ampoule into my prepared 50 ml of distilled water to make our stock 2%. I have no financial interest in this company but am a satisfied customer for many years. Phone number is: (800) 523-5874. Rosemary Walsh, EMF for the Life Sciences PSU
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
David Patton wrote: ======================================================== While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way? ========================================================== I am a bit surprised to hear that you are getting osmium tetroxide that is not in prescored ampoules. So far as I know, all of the major suppliers of EM chemicals, like SPI, have long since supplied their ampoules in the pre- scored form. The only time I have seen in recent years ampoules not in pre- scored form are those that are destined for other markets and applications.
Also, SPI, as well as the other major suppliers have long offered ampoule neck breakers, such as are described on URL http://www.2spi.com/catalog/tools/smtol21.html These are meant to be used one time and discarded. They are inexpensive and seem to provide an extra element of safety.
Using only prescored ampoules and an ampoule neck breaker might be easier on the nerves.....
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
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Use a triangular metal file to score around the neck, then use the usual precautions (protect fingers, open in hood , etc.) to pop open the vial. The EMS brand I use breaks open easily by wrapping paper towel/ gauze around top and gentle application of pressure.
Becky Garrison I am not an employee of any EM supplier.
-----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Friday, September 07, 2001 5:31 AM To: Monson, Frederick C. Cc: 'Microscopy Listserver'
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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Fred: Have you given any though to the fact that Coriolis' forces are different in the southern hemispher? Water drains from a basin in a counterclockwise motion south of the Equator and clockwise north of the Equator. Maybe wiggles and waggles are affected thesame way between the eastern and western hemispheres.
Regards,
Sam Purdy
Technical Center National Steel Corp. 1745 Fritz Dr. Trenton MI Voice:734-676-2682 FAX: 734-676-2682 spurdy-at-nationalsteel.com
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Karen:
Another source that we have come across is sodium phosphate buffer crystals. If fixation is carried out with sodium phosphate buffered soluitons, and then immediately transittioned to dehydration solvent (either EtOH or Acetone) then we have seen samples coated with fine crystals. We use either serveral water rinses between fixation and dehydration or an alternative buffer (i.e. Na Cacodylate)
} } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D. } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
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Gray, Peter, 1975, Microtomist's Formulary and Guide, R. Krieger, Huntington, NY. (Still available for the hardy bibliophile at http://WWW.WEB4U.COM/cgi-bin/full_page?0-88275-247-2) reports FOUR fixative mixtures that are attributed directly to Champy:
My favorite, of course, is number 2!!! whose last component provides a wonderful bit of chemical history. We see how Bayer must have started. Dupont.
Regards, and I know that this doesn't help at all!
But it is all part of the art.
Fred Monson
} ---------- } From: Richard Edelmann } Reply To: edelmare-at-muohio.edu } Sent: Wednesday, September 5, 2001 4:24 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Bio. Fixation: What is Champy's Fluid? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed } by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
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I am investigating EDS systems to integrate with a JEOL-6100 SEM. I'm leaning towards PGT's Spirit system. Other considerations are Oxford's INCA and Thermo Noran's Vantage. Does anyone have particularly positive or negative experience with PGT? I don't often hear much about that company. Any comments about the other possibilities would be appreciated, as well.
Thanks!
-Peggy
Peggy McKarns Macatangay, PhD Project Analyst 2 Celanese Corpus Christi Technical Center
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (john_bruss-at-bose.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 22, 2001 at 17:00:52 ---------------------------------------------------------------------------
Email: john_bruss-at-bose.com Name: John Bruss
Organization: Bose Corp.
Education: Graduate College
Location: San Diego, CA, USA
Question: What materials and process would an amateur use to suspend a spider in a clear cube for unmagnified artistic and historic use? Where are those materials available in small quantities?
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I am pretty sure that Alwyn is not talking about x-ray mapping. If that was all he wanted, I suppose the key disk is all that would be needed if he already has the other imaging applications and hardware installed.
Someone suggested using Auto to scan an image for multiple spectra. Indeed, the software and hardware should allow it. The spectra might then be exported and processed by some other package to perform the type of component analysis that has been described at MSA over the last few years. My current ISIS x-ray application (version 3.32) has the option of storing the x-ray data in single column MSA format. Previous versions placed multiple channels on a single line.
One hitch would be the limitation on the number of spectra per job. I think there is a limit of 1000 or so built into the database used for managing the data. I suppose it could be changed to another number, but we bumped into it some time back.
That would mean you could do a 32x32 raster of points saving a spectrum at each. A four second acquisition per point would mean a bit more than an hour for acquisition. That would not be too bad, but the spatial resolution would probably be too low to be very useful. But the data should readily fit onto hard drives with capacities of a few gigabytes.
I would be interested in seeing how the data processing is handled. If someone makes it work, I would be very interested in hearing the details.
Warren
At 10:33 AM 8/23/2001 -0400, you wrote: } -----------------------------------------------------------------------. } This is correct, the keydisk is all that is required for } the software to be enabled. However, there may be hardware } required, too, at least a cable interface (RS232?) to the } microscope. I believe the ISIS Autobeam likes to select } its own scan rates. You may need to speak with Oxford re: } compatibility with your particular microscope. } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Wednesday, August 22, 2001 9:23 PM, Fred Pearson } [SMTP:eoptics-at-mcmail.cis.mcmaster.ca] wrote: } } } } Alwyn: } } } } By spectral images, do you mean xray mapping? A "key" disk for the ISIS } } software program is required to activate the xray mapping facility. Of, } } course this will cost a bit of money to purchase from Oxford. When the } } ISIS program is loaded initially, all the software required to use the } } program is there, but activating the unpaid for parts of the program is } } the thing. } } } } Fred } } } } On Wed, 22 Aug 2001, Alwyn Eades wrote: } } } We have an Oxford ISIS EDS system. As it stands it is not set up to } } } acquire spectral images. We would like to be able to do spectral image } } } acquisition on this machine. Is there anyone out there who knows how } } } the system may be modified (macros, interfaces, whatever) to do it? We } } } do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that } } } helps. } } } .......... } } } Alwyn Eades } } } Department of Materials Science and Engineering } } } Lehigh University } } } 5 East Packer Avenue } } } Bethlehem } } } Pennsylvania 18015-3195 } } } Phone 610 758 4231 } } } Fax 610 758 4244 } } } jae5-at-lehigh.edu
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Mike, My guess is that either will do the trick, but I would take EDX as a shortened version of Energy Dispersive Analysis of X-rays, which, of course, is EDAX. I've also seen EDXA which would keep you out of trouble with EDAX. One really is doing a specific type of analysis, spectroscopy, which would cause me to lean towards Energy Dispersive Spectroscopy.
Ken Converse owner Quality Images third party SEM service Delta, PA
Ingram, Mike wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Not a serious question here, more of a curiosity. } } I use EDS when talking about x-ray analysis. I see many using EDX. Which } is correct? } } Mike Ingram } } }
Dear Listers, In a clear out of a cupboard I have found a pair of Debye-Sherrer cameras and accessories. If anyone would like them, please contact me - otherwise they're skip fodder. I used to demo this as an undergraduate lab some years ago, but I guess no-one uses them any more?
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone." Marconi Optical Components Limited Registered in England No. 4113798 Registered Office: One Bruton Street London W1J 6AQ
The CCD camera (model 679) in our CM20 TEM does not work any more. It is the fist commercial CCD camera of Gatan. The company is experiencing hard time to find the right replacing part. A programmable chip ( TEK1B2FV9 made by ACTEL) for the camera controller is needed. Then they can possibly figure out what problems with the camera.
If you have this part or any information which can help us to sole this problem please let me know. I can be reached at (613) 990-1403 or dashan.wang-at-nrc.ca
Try Prudence Rice's book Pottery Analysis: A Sourcebook (1987). This was the text on archaeological ceramics a few years back, and would be a good starting point. It is available for $65 on amazon.com.
Dear Katelyn, I will only add to Nelson Conti's remarks that you should be able to buy "blood-agar plates," which are Petri dishes already filled with a sterile, general bacterial culture medium, as well as sterile cotton-tipped swabs, from a medical supply store in your area. My daughter did a project along the same lines as yours and that's where we got the supplies. The dishes came in packs of 10, I think and they only cost a few dollars. We kept the inoculated dishes in a warm place. Around body temperature (37 degrees C, 98-99 degrees F) is ideal is you can find such a place. If you find bacterial colonies that have a clear ring around them, instead of the red of the rest of the blood agar, that is an indication that the bacteria in that colony are pathogenic (disease-producing). Good luck, and have fun.
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
At UIC we are less than 800m from the Blue Line of the cta L. When the lab was set up we bought an EMF cancellation system, in case. However, it turned out we did not need it despite doing high resolution annular dark field STEM with sub 0.2nm resolution.
While I was working for VG we did install a HB501 STEM in Halle, Germany. Here it was a different story. The main streetcar line through the city ran less than 100m from the site and to make matters worse a main feed for the system ran perpendicular to the line along a second wall of the building (probably 50m away). You could see each and every streetcar entering the stretch of line powered by the feed, it caused a sideways image jump! This was fixed by putting a Hall effect coil in the room at microscope bench level with a feed into the alignment coils.
If this is a new transit system then you are likely to suffer less problems with vibration and magnetic fields than you are with an old system where maintenance and standards of construction are likely to be poorer.
Alan
At 12:05 PM 9/7/2001 -0700, John C. Wheatley wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Make sure that you withdraw the specimen from the polishing solution as quickly as possible (!) and rinse immediately (!). I don't know what you are using to rinse your specimens. If your electropolishing solution is based on methanol, than it is a good idea to rinse it in methanol. I normally rinse my specimens twice in two different beakers with methanol or ethanol. Good luck!
"yan.hu" {yan.hu-at-brunel. To: microscopy-at-sparc5.microscopy.com ac.uk} cc: Subject: ask help
09/11/01 03:35 AM
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Dear Listers,
I got a problem when preparing TEM foils of Al-alloy by means of electro-polishing. A dark film produced on the specimen surface when it was been polishing. I think, the polishing condition is right and listed below:
Solution, 25 % Nitric acid plus 75 % Mechenol Temperature, about -30 degree C Voltage, 15-20V
I would be pleased to hear from anyone who has an idea to sort out this problem.
This is the second round. I have not received enough responses to give any meaningful data. If you are interested in the results of this survey, but have not responded for your facility, please do so. Thanks!
This is a survey that was passed out at the Facility Managers Meeting at this year's M&M meeting. We've only gotten 6 responses so far, and many more are needed for valid results (although obviously only one answer per facility). Please take a couple of minutes to fill this form in, and email back to me. Thank you!
Phil
MICROSCOPY FACILITY SUPPORT
This survey will help compile the nature and amount of support provided to microscopy facilities by universities, colleges, and research institutes. We would like to determine how much facility support is provided by the institution, and how much must be generated by user fees or other sources, on a per cent basis.
If we get enough responses to have valid data, we will post the results on the microscopy listserver and send them to the MSA bulletin. Names of institutions will not be included in the reported results.
Institution: Location: Nature of institution (Academic, Commercial, private research, etc.)
Type of Facility (institution core, satellite, departmental, etc):
(Place an 'X' by the appropriate answer) Predominately: Biological Materials Both
User base: Service Multi-user Both
Type of microscopes: SEM FESEM ESEM or LV TEM FETEM Major optical (Specify) Scanning Probe (specify) Other (specify)
Approximately what *per cent* of the funds for each of the following is provided by A) your institution, B) user fees, or C) grants
Salaries: A) B) C)
Instrument Maintenance (Service contracts, instrument insurance, etc): A) B) C)
Replacement of existing equipment: A) B) C)
Purchase of new equipment (not replacement for but addition to existing equipment): A) B) C)
Supplies: A) B) C)
Other operating expenses: A) B) C)
Are there other microscope facilities at your institution? if so, please indicate approximate number having electron microscopes. Ideally we would like a form filled out for each major facility (multiple instruments) at your institution if there are more than one.
Please include additional information such as novel funding ideas for your facility or for others at your institution.
Please email your responses to Philip Oshel peoshel-at-facstaff.wisc.edu -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Many thanks to everyone who responded to my question about electropolishing. The details of my experiment are: 1. it is an Al-Mg-Si alloy. 2. it is a twin jet-polish by Tenupole 3. The flow rate I used was 6. 4. The black film possibly appeares at the beginning of the polishing not after the foil prepared. 5. I noted that the current is more than 500 mA this time, not about 100 mA as before. This is a difference. Is it due to the problem of the instruments.
Has anyone done serial sections of decalcified mouse ears? I have been given some that were decalcified in EDTA for about 10 days, post-osmicated, dehydrated, infiltrated and embedded in Spurr resin. A grad student did the orienation for embedding, but along what plane I do not know. Each orientation is a little different. Can anyone recommend a consistent method of embedding?
This message is not relevant to our normal topics of discussion but I hope that under the circumstances you, Nestor, and others will forgive me for this once-off breach of the rules.
I am sure I speak on behalf of all subscribers to the MSA listserver when I express heartfelt shock and indignation at yesterdays tragedies in New York City, Washington DC and near Pittsburg. Those of us from elsewhere in the world assure our colleagues in the US that we share your grief, and offer our support at this difficult time. We extend our sincere condolences to those who have lost family members, friends and colleagues and we wish a speedy recovery to those who have been injured.
God bless, vasbyt and sterkte*.
Rob
* loosely translated = "hang in there and be strong"
======================================================= Robin H Cross Director : Electron Microscopy Unit, and Chairman : 15th International Congress on Electron Microscopy (ICEM-15) Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm http://www.icem15.com =========================================================================== Remember that ICEM-15 takes place in Durban, South Africa in September 2002 ===========================================================================
I have watched this conversation with interest and must admit that I am a very nervous about a student in a High School isolating pathogens on blood agar at 37 deg C. This pre-supposes that there are trained microbiologists, safe handling facilities and disposal methods for unknown human pathogens.
If someone was to perform a risk assessment on this activity would they even consider letting an unsupervised first year degree student isolate unknown pathogens in the microbiology department?
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Jill Verlander Reed wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Katelyn, } I will only add to Nelson Conti's remarks that you should be able to buy } "blood-agar plates," which are Petri dishes already filled with a } sterile, general bacterial culture medium, as well as sterile } cotton-tipped swabs, from a medical supply store in your area. My } daughter did a project along the same lines as yours and that's where we } got the supplies. The dishes came in packs of 10, I think and they only } cost a few dollars. } We kept the inoculated dishes in a warm place. Around body temperature } (37 degrees C, 98-99 degrees F) is ideal is you can find such a place. } If you find bacterial colonies that have a clear ring around them, } instead of the red of the rest of the blood agar, that is an indication } that the bacteria in that colony are pathogenic (disease-producing). } Good luck, and have fun. } } Jill Verlander Reed, D.V.M. } Associate Scientist } Director, College of Medicine Electron Microscopy Core Facility } University of Florida } P.O. Box 100215 } Gainesville, FL 32610 } verlaj-at-medicine.ufl.edu } Phone: (352) 846-0820 } Fax: (352) 846-3299
=============================================================== ORIGINAL MESSAGE:
Email: sbcook-at-bcpl.net Name: Katelyn Cook
Organization: St. Michael the Archangel
Education: 6-8th Grade Middle School
Location: Baltimore, MD
Question: I am planning to do a science fair project about germs and the importance of hand washing. I want to collect germs from common public places such as doorknobs, handrails, telephones, sink faucets, etc. What is the best way to collect the germs? Will I need to grow the germs in a petri dish after I collect them? Will I be able to identify different types of germs using my microscope?
Hi everyone! I wonder if this is the right place for my doubt... I need if someone could tell me whether a major and important difference exists between Paraformaldehyde 4% vs Formaldehyde4% as a fixative for brain tissue. Thank you :) Valeria Burgos
_________________________________________________________ ¿Lo probaste? Correo gratis y para toda la vida en http://correo.yahoo.com.ar
I embed and section mouse cochleas on a regular basis (anywhere between 1 and 4 microns in thickness). Is your tissue dissected down to the spiral-shaped bone of the cochlea itself? Do you want sections along the plane of the modiolus (the center columnar structure of the cochlea) so that you get a cross-sectional view of three to five organ of Corti profiles? The cochleas I embed still have some vestibular structures remaining.
If so, I embed the cochleas in flat silicone molds with the flat surface of bone (which faces the brain) facing the bottom of the mold and the spiral surface up. After the resin has been polymerized and the blocks removed from the mold, I remount the block onto flat-ended cylinder blanks. (These are made by closing the cap of a BEEM capsule, cutting off the pyramidal tip, placing them with the closed cap down, filling them to the rim with resin, polymerizing and removing the BEEM capsule.) The block is remounted (I usually use SuperGlue) onto the blank with the cochlea side down (the curved surface now faces down and the flat bony surface faces up). Excess resin is removed by trimming the block as usual. When I'm cutting into the block, I continually check the orientation. I know I'm close the proper orientation if I start to get profiles of the maculae and/or cristae in the vestibular structures and a base-to-apex view of the modiolus. Continue to adjust until you get the desired orientation.
I hope this gets you started.
Jaclynn M. Lett, Research Technician III jlett-at-cid.wustl.edu
Fay and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314-977-0257 fax: 314-977-0030
-----Original Message----- } From: stacey andringa [mailto:andrina-at-email.uc.edu] Sent: Tuesday, September 11, 2001 1:52 PM To: Microscopy-at-sparc5.microscopy.com
Has anyone done serial sections of decalcified mouse ears? I have been given some that were decalcified in EDTA for about 10 days, post-osmicated, dehydrated, infiltrated and embedded in Spurr resin. A grad student did the orienation for embedding, but along what plane I do not know. Each orientation is a little different. Can anyone recommend a consistent method of embedding?
We have been working with hemagglutination tests lately (something new for our EM lab), and are looking for a quick and easy way to score fields captured using a standard LM with a digital camera.
We have tried some of the image analysis packages available, but have not found them very easy to use. Also we have some questions about the images themselves - how many blood cells have to be stuck together before it's considered to be a clump? A clump is a 3 dimensional shape, so this has to be taken into account, doesn't it?
If anyone is doing this routinely and wouldn't mind giving us a few hints (or references), we'd really appreciate it.
Thanks in advance for any help you can give.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
The term of "Spectral Imaging" is not well defined yet. If it means the distribution of certain number of elements on the surface of sample, that is the X-ray mapping. If it means all of possibility on the element table (means energy spectrum) on the entire surface of sample (mapping), I do not think the Oxford ISIS can do it. The simple way is to use the "Area Analysis" in stead of point analysis. That will give you all element signature on the selected area.
In ISIS 300, there is an EBSD package attached (need to get key, again), this package can do the similar thing. It collects the electron backscatter diffraction pattern (EBSP) on a single point and compares it with a signed crystallographic pattern (one needs to be known and signed). It does this on an array point by point and displays the matrix as a map. The final result shows the contrast of crystallography on the surface layer ( {50nm). This can be called as the imaging of diffraction property of given crystal. It is a very good tool for the study of grain size and boundary.
It is up to user's interest, what is your goal for "spectral imaging". ISIS has another software package, call Particle Analyzer (Gun shot analyzer). Again, user needs to give information about the most possible present elements, and the software can do the rest to see if these elements are shown up. Without these pre-defined information, software will have no direction to go and have no goal to be achieved.
Regards,
Zhiyu Wang Maxtor Corp.
-----Original Message----- From: Zhiyuw [mailto:zhiyuw-at-home.com] Sent: Tuesday, September 11, 2001 9:51 PM To: zhiyu_wang-at-maxtor.com Subject: Fw: Spectral Imaging with ISIS
----- Original Message ----- From: "Warren E Straszheim" {wesaia-at-iastate.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, August 23, 2001 10:32 AM Subject: RE: Spectral Imaging with ISIS
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am pretty sure that Alwyn is not talking about x-ray mapping. If that was } all he wanted, I suppose the key disk is all that would be needed if he } already has the other imaging applications and hardware installed. } } Someone suggested using Auto to scan an image for multiple spectra. Indeed, } the software and hardware should allow it. The spectra might then be } exported and processed by some other package to perform the type of } component analysis that has been described at MSA over the last few years. } My current ISIS x-ray application (version 3.32) has the option of storing } the x-ray data in single column MSA format. Previous versions placed } multiple channels on a single line. } } One hitch would be the limitation on the number of spectra per job. I think } there is a limit of 1000 or so built into the database used for managing } the data. I suppose it could be changed to another number, but we bumped } into it some time back. } } That would mean you could do a 32x32 raster of points saving a spectrum at } each. A four second acquisition per point would mean a bit more than an } hour for acquisition. That would not be too bad, but the spatial resolution } would probably be too low to be very useful. But the data should readily } fit onto hard drives with capacities of a few gigabytes. } } I would be interested in seeing how the data processing is handled. If } someone makes it work, I would be very interested in hearing the details. } } Warren } } At 10:33 AM 8/23/2001 -0400, you wrote: } } -----------------------------------------------------------------------. } } This is correct, the keydisk is all that is required for } } the software to be enabled. However, there may be hardware } } required, too, at least a cable interface (RS232?) to the } } microscope. I believe the ISIS Autobeam likes to select } } its own scan rates. You may need to speak with Oxford re: } } compatibility with your particular microscope. } } } } Matt } } } } Matthew J. Lynn } } Center for Advanced Microscopy } } University of Miami } } (305)284-4736 } } mlynn-at-miami.edu } } } } } } On Wednesday, August 22, 2001 9:23 PM, Fred Pearson } } [SMTP:eoptics-at-mcmail.cis.mcmaster.ca] wrote: } } } } } } Alwyn: } } } } } } By spectral images, do you mean xray mapping? A "key" disk for the ISIS } } } software program is required to activate the xray mapping facility. Of, } } } course this will cost a bit of money to purchase from Oxford. When the } } } ISIS program is loaded initially, all the software required to use the } } } program is there, but activating the unpaid for parts of the program is } } } the thing. } } } } } } Fred } } } } } } On Wed, 22 Aug 2001, Alwyn Eades wrote: } } } } We have an Oxford ISIS EDS system. As it stands it is not set up to } } } } acquire spectral images. We would like to be able to do spectral image } } } } acquisition on this machine. Is there anyone out there who knows how } } } } the system may be modified (macros, interfaces, whatever) to do it? We } } } } do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that } } } } helps. } } } } .......... } } } } Alwyn Eades } } } } Department of Materials Science and Engineering } } } } Lehigh University } } } } 5 East Packer Avenue } } } } Bethlehem } } } } Pennsylvania 18015-3195 } } } } Phone 610 758 4231 } } } } Fax 610 758 4244 } } } } jae5-at-lehigh.edu } } }
Wageningen University Dept Agrotechnology and Food Sciences
Postdoctoral Research Position in Cell Wall Microscopy
A vacancy for a Postdoctoral Position is available in the area of microscopy and functionality of cell walls as part of the research program of Dr. Henk Schols, Laboratory of Food Chemistry. The successful candidate will be studying serum separation in tomato products and the influence of processing at the cell wall level. Ideally immunolocalization techniques will be used to localise specific polysaccharides in these samples.
Candidates must originate from outside The Netherlands and hold a recent Ph.D. in a related field. The appointment is for 9 months and is available immediately.
For more information, interested candidates should contact:
Dr. Henk Schols Wageningen University Dept Agrotechnology and Food Sciences Laboratory of Food Chemistry Bomenweg 2 6703 HD Wageningen The Netherlands
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Here is Washington, DC things are starting to get back to normal. Though what is normal has now changed. Our city was shut down for one day by cowardly terrorist acts. We are back at work and are carrying on. We will not let this break our spirit. The "Sleeping Giant" has been aroused and it is much larger than the perpetrators of this act realized as the Giant includes the global community.
Courage,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
} } } "R. Cross" {r.cross-at-ru.ac.za} 09/12/01 03:19AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear friends and colleagues
This message is not relevant to our normal topics of discussion but I hope that under the circumstances you, Nestor, and others will forgive me for this once-off breach of the rules.
I am sure I speak on behalf of all subscribers to the MSA listserver when I express heartfelt shock and indignation at yesterdays tragedies in New York City, Washington DC and near Pittsburg. Those of us from elsewhere in the world assure our colleagues in the US that we share your grief, and offer our support at this difficult time. We extend our sincere condolences to those who have lost family members, friends and colleagues and we wish a speedy recovery to those who have been injured.
God bless, vasbyt and sterkte*.
Rob
* loosely translated = "hang in there and be strong"
======================================================= Robin H Cross Director : Electron Microscopy Unit, and Chairman : 15th International Congress on Electron Microscopy (ICEM-15) Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm http://www.icem15.com =========================================================================== Remember that ICEM-15 takes place in Durban, South Africa in September 2002 ===========================================================================
I work on the University side of a large Medical center and yesterday after the attack in New York and Washington we received a broadcast e mail from the administration asking for blood. A couple of hours later I got to the blood bank and found a huge mob of medical workers in line to donate blood. Today the line was more then three hours long. I am going to wait a couple of days before donating blood. I think its realalistic that the demand for blood will be very high for at least a few weeks to come. I urge all healthy adults to join me in donating blood as its the only real thing that private citizens can do to help in this unprecedented catastrophe. Sincerely, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 JAH 1020 Locust Street Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cell Timothy.Schneider-at-Mail.TJU.edu
There are a couple of ways to reduce the orientation variation. One way is to use flat-bottomed Beem capsules for embedding molds (I haven't got the order info on these with me, but I can get it to you later, or you may get a reply from someone who has the info handy). These molds have a small enough surface area at the bottom that you can place the bulla or inner ear in them, say medial side down and they don't rock or float out of position. (Fill the mold before manipulating the specimen.)
Another way I have for embedding the cochlea is to split it in half, down the modiolus just prior to embedding and place the cut surface face down in the molds. I use half of a double edged razor blade to split it under a dissecting scope and I orient the cut from the apex to the base with the round window membrane perpendicular to the top surface. If you process the cochlea to this point before you split it, it is much firmer and tougher to work with and you won't lose as much tissue, but you do lose some. Hope this helps.
Karen Pawlowski, Ph.D.
stacey andringa wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Has anyone done serial sections of decalcified mouse ears? I have been } given some that were decalcified in EDTA for about 10 days, } post-osmicated, dehydrated, infiltrated and embedded in Spurr resin. A } grad student did the orienation for embedding, but along what plane I do } not know. Each orientation is a little different. Can anyone recommend } a consistent method of embedding? } } Stacey.Andringa-at-uc.edu
I find your comment interesting as I have volunteered to judge several Regional and National Science Fairs for high school aged students and have seen this particular experiment carried out by a number of the students. The students that I choose to judge are actually the younger group, who are "middle school" aged and I doubt the majority of them are getting much outside help, other than their science teachers or they would have picked a topic that was more unique (or geared to what the expert was doing).
I'm sure the petri dishes never leave the science lab. and the students were gloves while handling the cultures, but that is probably the extent of it....
Karen Pawlowski, Ph.D.
Malcolm Haswell wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have watched this conversation with interest and must admit that I am } a very nervous about a student in a High School isolating pathogens on } blood agar at 37 deg C. This pre-supposes that there are trained } microbiologists, safe handling facilities and disposal methods for } unknown human pathogens. } } If someone was to perform a risk assessment on this activity would they } even consider letting an unsupervised first year degree student isolate } unknown pathogens in the microbiology department? } } Malcolm Haswell } e.m. unit } School of Sciences } University of Sunderland } UK } } Jill Verlander Reed wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Katelyn, } } I will only add to Nelson Conti's remarks that you should be able to buy } } "blood-agar plates," which are Petri dishes already filled with a } } sterile, general bacterial culture medium, as well as sterile } } cotton-tipped swabs, from a medical supply store in your area. My } } daughter did a project along the same lines as yours and that's where we } } got the supplies. The dishes came in packs of 10, I think and they only } } cost a few dollars. } } We kept the inoculated dishes in a warm place. Around body temperature } } (37 degrees C, 98-99 degrees F) is ideal is you can find such a place. } } If you find bacterial colonies that have a clear ring around them, } } instead of the red of the rest of the blood agar, that is an indication } } that the bacteria in that colony are pathogenic (disease-producing). } } Good luck, and have fun. } } } } Jill Verlander Reed, D.V.M. } } Associate Scientist } } Director, College of Medicine Electron Microscopy Core Facility } } University of Florida } } P.O. Box 100215 } } Gainesville, FL 32610 } } verlaj-at-medicine.ufl.edu } } Phone: (352) 846-0820 } } Fax: (352) 846-3299 } } =============================================================== } ORIGINAL MESSAGE: } } Email: sbcook-at-bcpl.net } Name: Katelyn Cook } } Organization: St. Michael the Archangel } } Education: 6-8th Grade Middle School } } Location: Baltimore, MD } } Question: I am planning to do a science fair project about germs and } the importance of hand washing. I want to collect germs from common } public places such as doorknobs, handrails, telephones, sink faucets, } etc. What is the best way to collect the germs? Will I need to grow } the germs in a petri dish after I collect them? Will I be able to } identify different types of germs using my microscope? } } ---------------------------------------------------------------------------
To all our Amercian fellow microscopists and friends, The Australian Society for Electron Microscopy Inc sends its deepest heartfelt condolences to you all and your loved ones at this time of such tragic and unbelievable loss. Our warmest thoughts and prayers are with you all. Gerry Nash President ASEM Inc -- Geraldine (Gerry) Nash Electron Microscopist
EM Unit Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
Visit our EM Unit web site: http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp and Australian Antarctic Division web site: http://www.aad.gov.au/
To all our Amercian fellow microscopists and friends, The Australian Society for Electron Microscopy Inc sends its deepest heartfelt condolences to you all and your loved ones at this time of such tragic and unbelievable loss. Our warmest thoughts and prayers are with you all. Gerry Nash President ASEM Inc -- Geraldine (Gerry) Nash Electron Microscopist
EM Unit Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
Visit our EM Unit web site: http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp and Australian Antarctic Division web site: http://www.aad.gov.au/
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Thu, 13 Sep 2001, Gerry Nash wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all our Amercian fellow microscopists and friends, } The Australian Society for Electron Microscopy Inc sends its deepest } heartfelt condolences to you all and your loved ones at this time of } such tragic and unbelievable loss. } Our warmest thoughts and prayers are with you all. } Gerry Nash } President ASEM Inc } -- } Geraldine (Gerry) Nash } Electron Microscopist } } EM Unit } Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 } Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351 } } Visit our EM Unit web site: } http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp } and } Australian Antarctic Division web site: http://www.aad.gov.au/ } }
To all our American colleagues: On behalf of the Microscopical Society of Ireland, I would like to extend deepest sympathy and condolences to those who have been affected directly or indirectly by the atrocious terrorist attacks on the American Nation.
Sincerely, Alexander Black, President, Microscopical Society of Ireland
Department of Anatomy National University of Ireland, Galway.
There is a great difference. Paraformaldehyde is a POLYMER of formaldehyde(HCHO). Paraformaldehyde will yield HCHO if you heat the dry powder to around 90 degrees C (NOT RECOMMENDED!!!). I am sending a document under separate cover that will explain, but briefly, there are three words that are important here. Formalin (commercial, 40% HCHO solution in HOH, that contains from 5-15% methanol to prevent the HCHO from polymerizing), Paraformaldehyde (the polymer, a white powder), and formaldehyde solutions (generated from paraformaldehyde suspensions in water or buffer, which I designate as "pHCHO"). The attachment to the following email will explain in greater detail. Good question!!!
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I am looking for antibodies against double-stranded DNA, and eventually antibodies against histones. The two companies that I contacted, Biogenex and Pierce, discontinued its productions. Does anybody know for a supplier?
Tea
*************************************** Tea Meulia Research Scientist and Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
Dear All, We have had a last minute change in the program for our program for Microscopy and Digital Imaging on Sept. 27, 2001
1:15 "Imaging Tissue Morphogenesis in Zebrafish Embryos." Mark Cooper, University of Washington
Regards, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax **************************************************************************
C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
In regard to what is "appropriate", I share my own view.
I agree that science and this list server under these circumstances should be divorced from any shade of political statement.
The science community (emphasis on the community) is not divorced from emotion, thank goodness.
I believe we graciously accept the simple gestures of sympathy and not imply insult or rejection.
Justification: It is a historical, if not still surreal and forever infamous, time for world civilization.
With best regards and appreciation, Ed
Angela Klaus {avklaus-at-amnh.org} on 09/13/2001 05:07:53 AM
To: Gerry Nash {gerry.nash-at-antdiv.gov.au} cc: microscopy-at-sparc5.microscopy.com
Thank you, Gerry.
Best regards,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Thu, 13 Sep 2001, Gerry Nash wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all our Amercian fellow microscopists and friends, } The Australian Society for Electron Microscopy Inc sends its deepest } heartfelt condolences to you all and your loved ones at this time of } such tragic and unbelievable loss. } Our warmest thoughts and prayers are with you all. } Gerry Nash } President ASEM Inc } -- } Geraldine (Gerry) Nash } Electron Microscopist } } EM Unit } Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 } Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351 } } Visit our EM Unit web site: } http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp } and } Australian Antarctic Division web site: http://www.aad.gov.au/ } }
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Listers, About a year ago there was a discussion about using ammonia to clean Wehnelts. I dutifully saved all the emails because the technique looked interesting. Of course, when I went looking for these emails I couldn't find them. I looked on the list archives but was not able to come up with anything. Could someone who uses this technique contact me off list?
Thanks -- Glenn
=============================================================================== Glenn Poirier 3450 University St, rm. 238 MicroAnalytical Laboratory Montreal, Qc Earth and Planetary Sciences tel (514) 398 6774 McGill University fax (514) 398 4680 email: glennp-at-eps.mcgill.ca http://castaing.eps.mcgill.ca
++ Millenium hand and shrimp ++ ===============================================================================
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Thank you, all, for your support. I would like to share an article I received with everyone, especially my fellow Americans. I hope nobody is offended by it, for I do not intend, or wish, for that to happen.
Darrell
} I know you are all receiving a lot of emails like this today...but read } on: } } Subject: This, from a Canadian newspaper, is worth sharing. } } America: The Good Neighbor. } } Widespread but only partial news coverage was given recently to } remarkable editorial broadcast from Toronto by Gordon Sinclair, } a Canadian television commentator. } What follows is the full text of his trenchant remarks as } printed in the Congressional Record: } } "This Canadian thinks it is time to speak up for the Americans as } the most generous and possibly the least appreciated people } on all the earth. } Germany, Japan and, to a lesser extent, Britain and Italy were } lifted out of the debris of war by the Americans who poured in } billions of dollars and forgave other billions in debts. } None of these countries is today paying even the interest } on its remaining debts to the United States. } } When France was in danger of collapsing in 1956, it was the } Americans who propped it up, and their reward was to be insulted } and swindled on the streets of Paris. I was there. I saw it. } } When earthquakes hit distant cities, it is the United States } that hurries in to help. This spring, 59 American communities } were flattened by tornadoes. Nobody helped. } } The Marshall Plan and the Truman Policy pumped billions of dollars } into discouraged countries. Now newspapers in those countries are } writing about the decadent, warmongering Americans. } } I'd like to see just one of those countries that is gloating over } the erosion of the United States dollar build its own airplane. } Does any other country in the world have a plane to equal } the Boeing Jumbo Jet, the Lockheed Tri-Star, or the Douglas DC10? } If so, why don't they fly them? } Why do all the International lines except Russia fly American } Planes? } } Why does no other land on earth even consider putting a man or } woman on the moon? } You talk about Japanese technology, and you get radios. } You talk about German technology, and you get automobiles. } } You talk about American technocracy, and you find men on the } moon-not once, but several times-and safely home again. } You talk about scandals, and the Americans put theirs right } in the store window for everybody to look at. } } Even their draft-dodgers are not pursued and hounded. They are } here on our streets, and most of them, unless they are breaking } Canadian laws, are getting American dollars from ma and pa at home } to spend here. } } When the railways of France, Germany and India were breaking down } through age, it was the Americans who rebuilt them. } When the Pennsylvania Railroad and the New York Central went broke, } nobody loaned them an old caboose. } Both are still broke. } } I can name you 5000 times when the Americans raced to the help of } other people in trouble. Can you name me even one time when } someone else raced to the Americans in trouble? } I don't think there was outside help even during the San Francisco } earthquake. } } Our neighbors have faced it alone, and I'm one Canadian who is } damned tired of hearing them get kicked around. } They will come out of this thing with their flag high. } And when they do, they are entitled to thumb their nose at } the lands that are gloating over their present troubles. I hope } Canada is not one of those." } } Stand proud, America! } } I would hope that each of you would send this to as many people } as you can and emphasize that they should send it to as many } of their friends until this letter is sent to every person on the web.
Various methods for cleaning Wehnelt cylinders, and other internal parts of vacuum systems, are discussed in some detail in Section 2.10.4c (see bottom of page 72) of the book Vacuum Methods in Electron Microscopy (for info about this book see: http://www.2spi.com/catalog/books/book48.html and http://pup.princeton.edu/titles/6484.html).
Disclaimer: I make about $1.50 from each copy of this book sold. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
To all those people who have expressed their concerns,
I would like to thank all of those people who have sent their heartfelt messages and words of encouragement. Especially now in our time of need, it is uplifting to know all of our fellow microscopists from other nations are expressing their concern and support. During this time, when it is difficult to sort out our emotions, all of your words have helped to restore some of my faith in humanity.
Thanks you. Steve
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
I must humbly apologize for the "article" I sent. A coworker has informed me that it is a hoax. My first instinct was to not send it. I made the mistake of trusting the source, and allowing the video images of crowds of people cheering the deaths of tens of thousands of Americans in the matter of a few hours, cloud my better judgment.
I was, and am, sincere in my thanks for the support expressed on the list, and from all other sources.
Darrell
(These are my opinions. You can blame no one else for them.)
Usually we do our own SEM work, but a special job requirement may arise which I cannot accommodate using our equipment. It may be desirable to tensile fracture a thin ceramic sheet (predominately zirconia) and examine the fracture surface without breaking vacuum. EDS would likely be required as well. Magnification requirements *should* range from about 20x to 5000x. It would not surprise me, however, if 30-50 Kx might provide useful information.
If you have the equipment and take "outside" work, please contact me by direct email or phone.
Direct line to my office and lab: 434.522.6111 ...If A/C 434 fails, try the old one: 804
Thanks,
Woody White McDermott Technology, Inc. Lynchburg Research Center Lynchburg, VA
McDermott site: http://www.mtiresearch.com/ Personal site: http://woody.white.home.att.net
I am not sure is it original recipe or it's my modification, but anyway, the following works for me. I am sonicate Wehnelt cup in 1:1 diluted ammonia solution (stock ammonia solution I believe is 30%) for 10-15 min, rinse with deionized water and dry at +60 in the oven.
Good luck. Sergey
At 10:11 AM 9/14/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Wehnelt caps are usually made from stainless steel (304).
For JEOL wehnelts, I have had customers use "ammonium hydroxide hydrogen peroxide 30% by volume". I am not a chemist so I can't give you the particulars but this works great as it attacks everything except the stainless steel. I usually let the cap soak in this solution overnight.
Regards,
Earl weltmer ----- Original Message ----- } From: "Glenn Poirier" {glennp-at-eps.mcgill.ca} To: "Microscopy Listserver" {microscopy-at-sparc5.microscopy.com} Sent: Friday, September 14, 2001 7:11 AM
Dear American Fellow Microscopists, After the first moments of shock and grief let us express our deepest sorrow and sympathy to the American microscopists in particular and to the American people in general on behalf of all Hungarian microscopists. We have so many friends and colleagues there... To overcome these hard days and weeks let us wish you all strength both physically and in your soul, work and fight together for a better world! Kristof Kovacs
----------------------------------------------------------- Dr. Kristof Kovacs President, Hungarian Society for Microscopy University of Veszprem Veszprem, P.O.Box 158 H-8201 HUNGARY Phone: +36-(88)-421-684, +36-(30)-930-3931 Fax: +36-(88)-423-091
Food Structure & Functionality Symposium 2002 May 5 - 8, 2002, Palais des Congrès de Montréal o Montréal, Québec, Canada.
An international symposium leading Food Structure & Functionality studies through the 21st century "webaddress at the AOCS site (http://www.aocs.org/member/division/fsff/index.htm)"
Being held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
The symposium has two themes: * new and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Program Schedule Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl)
Monday, May 6th Morning
Opening of symposium Plenary Speaker Dairy Applications Session. Chairs: Mark Auty (mauty-at-moorepark.teagasc.ie) and Harjinder Singh (H.Singh-at-massey.ac.nz) Lunch break Afternoon Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl) and David Pechak (Dpechak-at-kraft.com)
Dedicated Poster Session Division Board Meeting
Tuesday, May 7th Morning Agricultural Applications of Microscopy and Imaging Session./ joint with Feed Microscopy Division. Topic/Tentative title (will probably be revised): New Microscopic Techniques for Identifying Food/Feed Constituents and Contaminants contacts: Mark Auty (mauty-at-moorepark.teagasc.ie) and Kim Koch
Lunch break: Division Luncheon and round table (expert) discussion. Topic TBA Afternoon Microbiology and Food Session. Chairs Judy Arnold (jarnold-at-saa.ars.usda.gov) and Ida Yates (iyates-at-ars.usda.gov)
Food Structure and Functionality Forum - Division Members Meeting
Wednesday, May 8th Morning Ingredients and Food Processing Session. Chairs: Diana Kittleson (dkittleson-at-pillsbury.com) and Bernhard Tauscher (bernhard.tauscher-at-bfe.uni-karlsruhe.de)
Lunch break Afternoon New Methods and Techniques for Food Structure and Functionality Analysis Session. Chairs: Kathy Groves Kgroves-at-LFRA.co.uk and Maud Langton maud.langton-at-sik.se
Closure of Symposium.
*-------------------------------------------------------------------------------------------------------------------- For further information, please go to the websites indicated, contact session chairs listed above, or myself.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Well, you hit the World Trade Center, but you missed America. You hit the Pentagon, but you missed America. You used helpless American bodies, to take out other American bodies, but like a poor marksman, you STILL missed America.
Why? Because of something you guys will never understand. America isn't about a building or two, not about financial centers, not about military centers, America isn't about a place, America isn't even about a bunch of bodies. America is about an IDEA. An idea, that you can go someplace where you can earn as much as you can figure out how to, live for the most part like you envisioned living, and pursue Happiness. (No guarantees that you'll reach it, but you can sure try!)
Go ahead and whine your terrorist whine, and chant your terrorist litany: "If you cannot see my point, then feel my pain." This concept is alien to Americans. We live in a country where we don't have to see your point. But you're free to have one. We don't have to listen to your speech. But you're free to say one. Don't know where you got the strange idea that everyone has to agree with you. We don't agree with each other in this country, almost as a matter of pride.
We're a collection of guys that don't agree, called States. We united our individual states to protect ourselves from tyranny in the world. Another idea, we made up on the spot. You CAN make it up as you go, when it's your country. If you're free enough. Yeah, we're fat, sloppy, easygoing goofs most of the time. That's an unfortunate image to project to the world, but it comes of feeling free and easy about the world you live in. It's unfortunate too, because people start to forget that when you attack Americans, they tend to fight like a cornered badger. The first we knew of the War of 1812 was when England burned Washington, DC, to the ground. Didn't turn out like England thought it was going to, and it's not going to turn out like you think, either.
Sorry, but you're not the first bully on our shores, just the most recent. No Marquis of Queensbury rules for Americans, either. We were the FIRST and, so far, only country in the world to use nuclear weapons in anger. Horrific idea, nowadays? News for you bucko, it was back then, too, but we used it anyway. Only had two of them in the whole world and we used 'em both.
Grandpa Jones worked on the Manhattan Project. Told me once that right up until they threw the switch, the physicists were still arguing over whether the Uranium alone would fission, or whether it would start a fissioning chain reaction that would eat everything. But they threw the switch anyway, because we had a War to win. Does that tell you something about American Resolve? } So who just declared War on us? It would be nice to point to some real estate, like the good old days. Unfortunately, we're probably at war with random camps, in far-flung places. Who think they're safe. Just like the Barbary Pirates did. Better start sleeping with one eye open. There's a spirit that tends to take over people who come to this country, looking for opportunity, looking for liberty, looking for freedom. Even if they misuse it. The Marielistas that Castro emptied out of his prisons, were overjoyed to find out how much freedom there was. First thing they did when they hit our shores, was run out and buy guns. The ones that didn't end up dead, ended up in prisons. It was a big PITA then (especially in south Florida), but you're only the newest PITA, not the first.
You guys seem to be incapable of understanding that we don't live in America, America lives in US! American Spirit is what it's called. And killing a few thousand of us, or a few million of us, won't change it. Most of the time, it's a pretty happy-go-lucky kind of Spirit. Until we're crossed in a cowardly manner, then it becomes an entirely different kind of Spirit.
Wait until you see what we do with that Spirit, this time. Sleep tight, if you can. We're coming. And when we arrive, well, let's just say it won't be pretty.
I have been given a piece of titanium that contains small precipitates of unknown composition and asked to prepare a TEM sample so that the precipitates can be analysed. As it is only the precipitates that are of interest I thought that the best approach would be to make an extraction replica. However, to do that need to find a way to etch away the titanium matrix while leaving behind the unknown precipitates.
I would be grateful for any recommendations for a suitable etchant that will preferentially remove the titanium matrix so that the extraction replica can be made.
Regards,
Martin Saunders. --
*****************************************
Dr. Martin Saunders, Lecturer, Centre for Microscopy and Microanalysis, University of Western Australia, Crawley, Western Australia 6009, Australia.
It has been very positive to see, on the microscopy listserver, messages from microscopy communities in other nations addressed to those of us who live and work in the United States offering sympathy for the tragic events of last Tuesday. These messages have, without exception, been measured, responsible and generous. We are grateful for them.
Speaking for myself, I want to say how offended I was to read, on the microscopy listserver, "Letter to a Terrorist" from Peter Jordan. There are no terrorists on this list server. This is not an appropriate place for this kind of polemic. I hope that those who take exception to Mr. Jordan's remarks will refrain from responding to the substance of what he says. The last thing we want is for the listserver to be taken over by a political debate on the issues raised by the attacks on New York and Washington.
Alwyn -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I like to dissolve fat in insects bodies what could be the best chemical . benzine, carbon tetrachloride, or???
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
Could anybody recommend the good method of sample preparation for SEM imaging on carbon nano tubes? The sample is a powder type including single wall carbon nanotubes (SWCNTs). Thank you very much in advance.
""} Subject: This, from a Canadian newspaper, is worth sharing. } } America: The Good Neighbor. } } Widespread but only partial news coverage was given recently to } remarkable editorial broadcast from Toronto by Gordon Sinclair, } a Canadian television commentator. } What follows is the full text of his trenchant remarks as } printed in the Congressional Record:""
Gordon Sinclair's piece was first broadcast in 1973 - see:
I'd like to thanks to all who has collaborate for the answers on tempers studies on potteries by SEM. I hope in brief get in touch with some of them.
I was trying to don't say a word on the terrorism subject, that because sometimes for me when something so horrible happens with me I prefer stay quiet on my side in order to recover my balance to overcome such pain. However, I would like to expreess my deep feelings to American people and gouvernement. Since the attacks I could not stop to think how terrible the human been can be, even if sometimes the streets can show us terrible facts. I've watched several chanels, brazilian and foreigners, and still can understand what hapens and why that people has died. I would like to remark my sympathy to your gouvernement which, in my oppinion, is doing their job in this difficult situation as well as possible. We hope you can find find a pacific way to solve this problem, however, if you can't don't mistake several other countries around the world, including Brazil, and I belive some arabian countries too, is supporting you and almost ready to stay beside you. You know the American spirit is not living only in US as well stand by Peter Jordan, this is the spirit that you can see in many other countries who respect the Democracy, The Human Rights, The Freedom, and have been proclamed in the France Revolution, US Constitution as well as in many other constitutions and UN Declarations. Again, here in Brazil we're deeply touched by your pain and we hope help you as better as possible to overcome this terrible incident.
The Ohio State University Microscopic and chemical Analysis Research Center (MARC) anticipates an opening for an electron microscopist beginning October 1 to operate a scanning electron microscope (SEM) and an electron microprobe (EPMA). The position includes working with researchers, teaching students the basic theory and practical SEM and EPMA measurement techniques, collaborating on research projects and directing staff assistants. We seek an excellent microscopist who enjoys the exciting atmosphere of a major research university and will enjoy collaborating with scientists on a broad variety of projects. The MARC serves the entire Ohio State University as well as other institutions, with projects from diverse research areas including geology, materials science, environmental sciences, biological and biomedical sciences, dentistry, textiles. The MARC focuses on elemental analysis using from microscale to bulk analysis using SEM, EPMA, inductively coupled plasma optical emission spectrometry and inductively coupled mass spectrometry with laser ablation or solution sampling. The MARC also teaches traditional or short courses on each of the techniques. The position is a full time, hard money funded job.
If you know of a suitable candidate for this position, please contact Dr. John Olesik, MARC Director, at olesik.2-at-osu.edu {mailto:olesik.2-at-osu.edu} or (614) 292-6954. Excellent candidates with a range of experience and potential to grow will be considered.
---------------------------------------------------------------------------- -- John Olesik Adj. Assoc. Professor, Research Scientist MARC Director Microscopic and chemical Analysis Research Center Ohio State University 125 S. Oval Mall 275 Mendenhall Laboratory Columbus, OH 43210
The "article" you forwarded is a hoax in that it was originally broadcast in 1973 and the author Gordon Sinclair died in 1984. However, many will agree that the ideas in the broadcast apply as well today as they did then.
We have a used Amray 1000A with a PGT IMIX EDS that we will part with, and are seeking best offer. The equipment details are on the used equipment page of the MSA web site.
I am currently recruiting for a Bristol (South West England) based high technology company which is seeking an experienced microscopist to join its pioneering team as a Lab Manager. I would like to know whether or not you might know of anybody suitably qualified who may be interested. My client will be happy to arrange Working Visa details. Regards.
Hello Haeseong, If you are just looking at the nanotubes, it would be easy to place some of the powder on double-sided carbon tape or double sided Cu tape, and just put the sample in the SEM. The important thing is to have the machine very well aligned, and astigmatism corrected for a small spot size and a voltage of about 5kV or less. You should start seeing useable information at a magnifcation range of 30-50kx and above.
-Brad
---------- From: Haeseong Lee Sent: Monday, September 17, 2001 5:55 AM To: Microscopy-at-sparc5.microscopy.com Subject: SEM imaging on CNT
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
?? Hello.
Could anybody recommend the good method of sample preparation for SEM imaging on carbon nano tubes? The sample is a powder type including single wall carbon nanotubes (SWCNTs). Thank you very much in advance.
I seriously doubt that Mr. Jordan meant or implied that there were terrorists in the Listserver.
Much like the lamentations I hear on the radio seemingly directed at terrorists about how the terrorists have "awakened a sleeping giant" etc. I doubt the author meant that everyone listening to the radio is a terrorist.
I am sure it was meant as a public message to vent all of our outrage at this terrible event.
Regards,
Earl Weltmer
----- Original Message ----- } From: "Alwyn Eades" {jae5-at-lehigh.edu} To: "EMNET" {microscopy-at-sparc5.microscopy.com} Sent: Monday, September 17, 2001 5:35 AM
I'm not familiar with recent reviews or specific insect protocols, but there's an older book by Morris Kates called techniques in lipidology -part of a series of lab -type manuals. One of the more universal ways to dissolve / extract lipids is the Bligh-Dyer method (Can J Biochem & Physiol 1959 37:911-917 , paraphrased from Kates:
Extract the specimen with one volume of chloroform:methanol:water 1:2:0.8, which efficiently extracts lipids. the extract is then diluted with one volume each of chloroform and water tp form the two-phased system, chloroform and Methanol/water (1.0:0.9). Any water-soluble contaminants are thus partitioned into the methanol-water phase.
Carbon tet is probably NOT the best to extract lipids with -they are more soluble in chloroform
Richard
} "Vr. Richard Bejsak-Colloredo-Mansfeld" {ricardo-at-ans.com.au} 9/17/01 5:37:30 AM } } } Dear Colleagues
I like to dissolve fat in insects bodies what could be the best chemical . benzine, carbon tetrachloride, or???
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
Hello folks. I am trying to fix and stain with lanthanum nitrate and have followed procedures in Hyatt's book. It mentions that lanthanum will precipitate in the cold, and when mixed with phosphate buffer. I have used cacodylate buffer, room temperature solutions and checked the pH of my buffer(7.2), but I am seeing a lot of ppt in all the solutions (glut fix, wash and osmium fix). I am wondering if anyone has any suggestions about how to alleviate this problem. Is the precipitate harmful to the tissue? Can I filter the solutions to get rid of big clumps? Is this normal? I would appreciate any feedback from someone who has used the stuff. Thanks, JoAnn
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Anyone taking down a darkroom or willing to donate a good photographic processor for black and white prints to a Children's Hospital?
-- Jacob Bastacky, M.D. Research Physician Children's Hospital Oakland Research Institute 5700 Martin Luther King Jr. Way Oakland, California 94609 Telephone: 510.450.7639 email: jbastacky-at-CHORI.org FAX: 510.450.7910
Senior Scientist, Visiting National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 sjbastacky-at-lbl.gov
I would like to hear some opinions about the Idefix EDS system, commercialized by SamX in France. Are they (probably french) List Members who know about it ? Please contact me off-list by mail.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Hope you are not knocked off from your normal life by the tragic events.
Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a regular sized room. Somehow we feel there is a bit above normal acoustic echos going on in the lab which interferes with the TEM. So we hope to find some kind of insulation to be put on the wall and/or around the TEM to improve the performance.
Any thoughts and suggestions are highly appreciated.
Thanks and take care.
********************************* Chaoying Ni Electron Microscopy Facility College of Engineering University of Delaware Newark, DE 19716 *********************************
Dear Cathy, When I was at the Australian Electron Microscopy Meeting there were a lot of papers about wool, including extensive TEM. Perhapps Mel Gibson or someone at CSIRO (www.csiro.au) could help. At 09:55 AM 9/17/01 +1000, you wrote: } Hi, } } Can anyone recommend a method for preparing wool from sheep for TEM? Is it } easy to section? } } Thanks } } Cathy Gillespie } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Dear Martin, Although I have not tried an extraction replica on Ti alloys myself, my Vander Voort lists several Ti alloy etchants, most based on combinations of HF asnd other strong mineral acids. The first is: "20 ml HCl 40 ml HF 40 ml H2O Immerse specimen in solution at 120 - 150 degrees F for 20-30 min. If smut forms immerse in 30% H2SO4 for 3 min." The simplest is: "50 ml HCl 50 ml H2O General-purpose macroetch (Ogden anbd Holden)." Hope this helps. At 06:40 PM 9/17/01 +0800, you wrote: } Dear all, } } I have been given a piece of titanium that contains small } precipitates of unknown composition and asked to prepare a TEM sample } so that the precipitates can be analysed. As it is only the } precipitates that are of interest I thought that the best approach } would be to make an extraction replica. However, to do that need to } find a way to etch away the titanium matrix while leaving behind the } unknown precipitates. } } I would be grateful for any recommendations for a suitable etchant } that will preferentially remove the titanium matrix so that the } extraction replica can be made. } } Regards, } } Martin Saunders. } -- Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a regular sized room. Somehow we feel there is a bit above normal acoustic echos going on in the lab which interferes with the TEM. So we hope to find some kind of insulation to be put on the wall and/or around the TEM to improve the performance.
Any thoughts and suggestions are highly appreciated.
Dear Chaoying, One of the other listers will surely know a company which produces sound-dampening material, but while you wait for it to be delivered, try using either styrofoam or bubble wrap as a temporary solution. The styrofoam peanuts would probably work best, but they will take a lot of labor to install--although you might be able to put up something in front of the walls and backfill with them. Cloth drapes also work to dampen sound. Furthermore, you only need to be concerned with the frequency range which matches resonances in your scope, so when you purchase the insulation, be sure it will absorb those frequencies. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Your comments about the European aviation industry are certainly true and I would expect that everyone on the list knows about the widespread use of the Airbus. Consequently, I did not think it was necessary to explain that I did not mean that the parts of the radio broadcast which include specific references to the "American technocracy" which are so clearly outdated should be considered as applicable today.
I completely agree with you that care must be taken regarding references to the Sinclair broadcast. In fact, in my original posting, I said "many will agree", which implies that "not all will agree". Also, I referred to the "ideas" of the broadcast, not the "specifics" of the broadcast. But as your e-mail points out, the ambiguity of the "ideas" can still lead to confusion, since they are left to personal interpretation.
Actually, I hesitated to respond to the original posting at all since it was clearly off-topic, however, the magnitude of the attack and the historical foundations of the "hoax", made me think that a short clarification regarding the original radio broadcast may be of general interest. I certainly did not mean to imply anything negative regarding the European (or any other country's) patriotism or spirit and I will be more careful regarding postings in the future that may have any nationalistic interpretations.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
For more information or to register, contact Lou Ross at rosslm-at-missouri or (573) 882-4777.
Central States Microscopy and Microanalysis Society Fall Meeting Friday, October 19, 2001 Adams Conference Center Veterinary Medicine Building University of Missouri Columbia, Mo 65211
8 - 8:45 Registration
8:50 Welcome
9:00 Tobias Baskin, University of Missouri-Columbia, "Ultrastructure of the Plant Cell Wall Analyzed with Field-Emission Scanning Electron Microscopy"
9:15 Nadia Navarrete-Tindall, University of Missouri-Columbia, "Morphology of the Endangered Legume Amorpha nitens Boynton and its Rhizobia Symbiont"
9:30 Heide Schatten, University of Missouri-Columbia, "Cytoskeletal Dynamics in Apicomplexan Parasites"
9:45 Joe Simmons, University of Missouri-Columbia, "Idiopathic Interstitial Pneumonia in Laboratory Rats"
10:00 Marty Katz, University of Missouri-Columbia, "Stem Cell Transplantation Therapy for Inherited Neurodegenerative Disorders"
10:15 Tom Phillips, University of Missouri-Columbia, "Fixation of Biological Tissues - Science or Art?"
10:30 - 10:45 BREAK
10:45 Robert Hall, Associate Vice Provost of Research, University of Missouri-Columbia, "The University of Missouri: Taking the Lead in National Research Funding"
11:00 Keynote speaker: Kenneth Moore, University of Iowa, MSA Tour Speaker, "Applications of Microscopy Techniques to the Study of Genetic Therapy Research"
12:00 - 1:30 Lunch
1:30 John Bozzola, SIU-Carbondale, "Multidisciplinary Approach to Teaching Electrion Microscopy"
1:45 Howard Wilson, University of Missouri-Columbia, "Creating a Scientific Poster Presentation using Microsoft Powerpoint"
2:00 Scott Walck, PPG, "Low Angle Cleavage - A New Specimen Preparation Method for TEM"
2:15 Louis Ross, University of Missouri-Columbia, "Exciting Times in Scanning Electron Microscopy and X-ray Microanalysis"
2:30 Vladimir Dusevich, University of Missouri-KC, "Practical ESEM"
2:45 Jeff Speakman, University of Missouri-Columbia, "Laser Ablation ICP-MS as a Tool for Characterization of Archaeological Materials"
3:00 - 3:15 BREAK
3:15 Cammy Bright, University of Missouri-Columbia, "A Laser Ablation ICP-MS and SEM Study of Rare Earth and Trace Elements in Conodonts"
3:30 Mike Amspoker, Westminster College, "A Light and Scanning Electron Microscopy Study of the Freshwater Diatom Desmogonium rabenhorstianum Grunow"
3:45 Heather Ramsay, University of Missouri-Columbia, "New Applications of SEM in Osteological Morphology Research"
4:00 Dickerson Moreno, University of Missouri, "Development of N-Type Diamond Semiconductor Through Field Enhanced Diffusion by Optical Activation"
4:15 Alejandro Suarez, University of Missouri, "A New Method to Fabricate Polycrystalline Diamond as a P-Type Semiconductor"
4:30 Closing Remarks
4:45 CSMMS Business Meeting
Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.missouri.edu/~geosclmr/ebaf.html
I've just had the misfortune to smash the glass bell jar on my Edwards 306 coater. About 305mm dia, 400mm high.
While I can and maybe should buy a glass replacement, I'm tempted to make one from a 400mm length of 305mm diameter stainless-steel pipe with a flat top of 10mm thick polycarbonate.
I've never liked having my face too close to the glass one when it's in use, anyway.
Anyone got any comments/recommendations?
Anyone ever had a glass one implode in use?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I have not known a glass tube to implode but we use an outer cover just in case (perspex tube). Tubes are checked for cracks before use and previous problems have been obvious, usually by the many bits all over the floor!
My worry would be the 10mm polycarbonate top, at 300mm dia that's over 1500 pounds (750Kg?) force. I would want something thicker or stronger.
I also like being able to look in the side to see my coating set up after evacuation and the evaporation (through appropriate goggles).
Regards, Ron
} I've just had the misfortune to smash the glass bell jar on my } Edwards 306 coater. About 305mm dia, 400mm high. } } While I can and maybe should buy a glass replacement, I'm tempted to } make one from a 400mm length of 305mm diameter stainless-steel pipe } with a flat top of 10mm thick polycarbonate. } } I've never liked having my face too close to the glass one when it's } in use, anyway. } } Anyone got any comments/recommendations? } } Anyone ever had a glass one implode in use? } } cheers } } rtch } } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
There are two sources of noise, externally and internally generated. You have specificcally asked about echos so I assume that your problems stem from the rotary pumps, cooling fans, etc. on equipment that is essential to have operating in the room.
Try to get the rotary pumps and power supply rack into an area that can be screened off from the instrument column but remember that it will still need air flow for cooling.
For the desk and computer fans use some blackout curtains around the walls. This is readily available and is usually black one side and coloured the other so you can have a nice Oxford blue showing into the room:-) Make sure that it is at least 1.5 lengths, preferably 2x - ie. 7.5m of curtaining is a minimum for a 5m wall 10m is best. If you get the rotary pump and power supply at the back of the room you can hide them behind 3 layers of the same curtaining.
We have found this very effective for our HREM instruments and our FEGTEM.
Noise from outside the room needs dealing with outside the room.
Regards, Ron
} Dear Listers, } } Hope you are not knocked off from your normal life by the tragic events. } } Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a } regular sized room. Somehow we feel there is a bit above normal acoustic } echos going on in the lab which interferes with the TEM. So we hope to } find some kind of insulation to be put on the wall and/or around the TEM to } improve the performance. } } Any thoughts and suggestions are highly appreciated. } } Thanks and take care. } } ********************************* } Chaoying Ni } Electron Microscopy Facility } College of Engineering } University of Delaware } Newark, DE 19716 } ********************************* } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
We have a 2010F in a similar sized room and we use floor to ceiling black curtaining - the heavier the better (ours are double thickness and lined) - and it makes a big difference. Hope this helps Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
Don't use a polycarbonate end plate to cover the end of your vacuum chamber. A flat plate of polycarbonate isn't strong enough to span the opening.
Edwards did sell glass cylinders with metal endplates on some of their 306 coaters--in particular their old ion beam sputtering system. The unit I have here has a 1/2 inch thick Aluminum plate spanning a 12 inch diameter glass vacuum cylinder. The glass cylinder is the same type of glass you find on a normal bell jar. It just has an L-gasket on both the top and bottom. Be careful that you use an adequate grade of aluminum for the top plate.
The advantage of the cylinder with metal top plate is that you can have feedthroughs to the vacuum chamber from the top as well as through the base plate.
I suspect that you can get the glass cylindrical glass vacuum vessels and L-gaskets from the usual vacuum supply houses.
cheers, Henk Colijn
At 12:32 PM 9/19/01 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
I have not known a glass tube to implode but we use an outer cover just in case (perspex tube). Tubes are checked for cracks before use and previous problems have been obvious, usually by the many bits all over the floor!
My worry would be the 10mm polycarbonate top, at 300mm dia that's over 1500 pounds (750Kg?) force. I would want something thicker or stronger.
I also like being able to look in the side to see my coating set up after evacuation and the evaporation (through appropriate goggles).
Regards, Ron
} I've just had the misfortune to smash the glass bell jar on my } Edwards 306 coater. About 305mm dia, 400mm high. } } While I can and maybe should buy a glass replacement, I'm tempted to } make one from a 400mm length of 305mm diameter stainless-steel pipe } with a flat top of 10mm thick polycarbonate. } } I've never liked having my face too close to the glass one when it's } in use, anyway. } } Anyone got any comments/recommendations? } } Anyone ever had a glass one implode in use? } } cheers }
Dear Ritchie,
I have a polycarbonate lid for a vacuum apparatus with the same diameter as your coater. It is about 45 mm thick, and it has not given me any trouble. I agree with Ron that 10 mm is not thick enough.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
We've had to contend with noisy rooms in nearly all of our electron microscope laboratories. Here is a list of things to do:
* Try to eliminate the source of the noise. * Call in the microscope manufacturer to provide a noise spectrum (dB versus Hz). Ask which frequencies are a problem (usually low frequencies). * Look for devices and materials which will attenuate the noise. Pay particular attention to their effectiveness in attenuating the problem frequencies.
Here are a few companies which manufacture items you may need: * Kinetics Noise Control, http://www.kineticsnoise.com * Industrial Acoustics Company, http://www.industrialacoustics.com * Industrial Noise Control, http://www.industrialnoisecontrol.com
I've worked several times with David Mitchell, VP at The Huff Co., Lake Bluff, IL (Chicago area), 1-847-362-7440, to ameliorate our noise problems. Such companies will evaluate the site, recommend solutions, and even install the various devices and materials from the manufacturers.
Some of the things we've done are: * glued "egg-crate" foam on the walls * installed fabric-covered fiberglass panels on the walls * installed duct silencers in air ducts * wrapped air ducts with fiberglass batting backed with "loaded vinyl" * hung fiberglass noise-absorbing curtains * installed isolation pads under vibrating equipment
If you install wall panels or foam, it is usually not necessary to cover all of the wall surfaces. There are lots of things to consider, so evaluate your site carefully.
..Russ Cook ---------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 recook-at-anl.gov
} -----------------------------------------------------------------------. } Dear Listers, } } Hope you are not knocked off from your normal life by the tragic events. } } Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a } regular sized room. Somehow we feel there is a bit above normal acoustic } echos going on in the lab which interferes with the TEM. So we hope to } find some kind of insulation to be put on the wall and/or around the TEM to } improve the performance. } } Any thoughts and suggestions are highly appreciated. } } Thanks and take care. } } ********************************* } Chaoying Ni } Electron Microscopy Facility } College of Engineering } University of Delaware } Newark, DE 19716 } *********************************
Is it common to use mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? Is a control using the secondary antibody (anti-mouse-IgG-gold-conjugate) alone sufficient to make sure that one is detecting specificly the primary antibody and not tissue-internal mouse antibodies? Does somebody know examples from the literature where people used mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? I am a botanist, so until now I never ran into that sort of problems.
Thanks,
Stefan
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer Universität zu Köln Botanisches Institut Gyrhofstr. 15 50931 Köln Germany °°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°
I didn't get to the MSA meeting this summer, but I heard through the grapevine about Gatan's new UltraScan 1000 and saw their poster. I was very impressed with the 4K x 4k montage taken at 5k mag. It looked publishable.I'd be interested in hearing comments/impressions from anyone who a chance to use it. Thanks,
Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
We are about to install a Tecnai 20 TEM. In addition to isolating rotary pumps and power supply as suggested by Ron Doole, we are covering the walls with Armstrong Sound Soak Panels.
Frank
At 11:25 AM 9/18/01 -0400, Chaoying Ni wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Is it common to use mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? Is a control using the secondary antibody (anti-mouse-IgG-gold-conjugate) alone sufficient to make sure that one is detecting specificly the primary antibody and not tissue-internal mouse antibodies? Or are there other things I can doo as a good control?
Does somebody know examples from the literature where people used mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? I am a botanist, so until now I never ran into that sort of problems.
Thanks,
Stefan
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer Universität zu Köln Botanisches Institut Gyrhofstr. 15 50931 Köln Germany °°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°
Hello to the person from down under with the broken bell jar:
I have been using a Denton 502A carbon evaporator for the past decade and have also had some visions of it imploding. The model I have has a metal mesh cover over the jar and while it would not stop little pieces of flying glass it would catch the big ones. As far as using a home made unit goes, I would be vary cautious. At the very least consult with some kind of structural engineer who is familiar with vacuum systems. Be careful. Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 JAH 1020 Locust Street Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cell Timothy.Schneider-at-Mail.TJU.edu
Sara, actually the total package for the UltraScan 1000 is about $100K. The UltraScan 4000 is the one you've quoted and that is a 4kx4k camera. The 1000 has the 2kx2k camera and montages the field requiring 4 exposures. I believe that is the way it works.
Hank
-----Original Message----- } From: Sara Miller [mailto:saram-at-duke.edu] Sent: Wednesday, September 19, 2001 1:58 PM To: Hank Adams
Dear Chaoying, One unlikely source of noise we recently found came from the diffusion pump on our CM 300. It was crackling like diffusion pumps due, but we could see our HRTEM image shake with the big crackles. The solution was deduced by our excellent FEI field engineer. It was to reduce the temperature in the diffusion pump by reducing the amount of oil charge. If you have noise problems that go snap, crackle, pop in the dark, try checking your oil. Ciao for now, Ken
Dear Chaoying, One unlikely source of noise we recently found came from the diffusion pump on our CM 300. It was crackling like diffusion pumps due, but we could see our HRTEM image shake with the big crackles. The solution was deduced by our excellent FEI field engineer. It was to reduce the temperature in the diffusion pump by reducing the amount of oil charge. If you have noise problems that go snap, crackle, pop in the dark, try checking your oil. Ciao for now, Ken
10 mm polycarbonate? I would suggest much thicker. Have no idea how much thick. I think the problem is that your pipe has a huge diameter. The force on your polycarbonate will be proportional to the size of area (1 kg/cm2 x 706 cm2=approx 700 kg, 1600 lb). I afraid, the total cost of the pipe etc would be comparable to the cost of Bell Jar. You may find Bell Jar on used equipment sale for about $300. Sergey
At 12:32 PM 9/19/01 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
This is incredible. I would suggest that this is a first EM digital camera which produces comparable to the film image quality. The chip is huge, so they have 80-90% area of the film on the chip. The images are very sharp. The technical problem there was to read information from such huge chip. They used new technology when chip (virtually?) splitted on quarters and read in parallel. They did a great job. The camera is very expensive by the way.
Sergey
At 11:00 AM 9/19/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I think you wrong. They do have two new cameras 2x2K and 4x4K. This is real number of pixels on the chip. I also believe, Ultrascan1000 is much, much more expensive than you suggest. As per our conversation with Gatan people, they expect to start selling that cameras at the end of the year, so it's very unlikely the price is known for now (I did not ask them directly about the price, but have impression that it's much more expensive that previous models, which were in the $70K range). Current Gatan's cameras (600W/CW Multiscan for instance) already has capacity for multiple exposure I believe. Actually, it's not a properties of camera but software. It's called "Auto-montage" and you could "montage" as many pictures as you want. By the way, Multiscan cameras has 2-4x (not sure in exact number) bigger area of view than film, which sometime is very useful. Over last few month I was studying EM digital cameras on the makket very hard, so if you have questions, may be I could help.
Sergey.
At 02:42 PM 9/19/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I would appreciate any advice on the following question. I am planning to do fluorescence microscopy and electron microscopy on the same sample. I would like to do immunolabelling on cells grown in a culture dish followed by plastic embedding of the same sample and further preparation for EM. In order to find the right area of cells to be sectioned for EM (previously detected by immuno fluorescence) I suppose one has to have some kind of grid on the culture dish. I am aware of the fact that there are gridded coverslips available, however, I always had a lot of trouble to remove the coverslip from the glass after embedding (used lN2 and so on). So it would be very helpful if anyone out there could provide any other working recipes, references for methods or sources for culture dishes with some kind of grid on them.
Dr. Andreas Brech EM-Unit, Dept. of Biology, University of Oslo. P.O.Box 1062 Blindern, Room U 139 Street address: Moltke Moes vei 32 N-0316 Oslo 3 Norway Tel.: + 47-22 85 47 25/61 89 (work) + 47-22 43 83 23 (privat) Fax.: + 47-22 85 47 26 e-mail.: abrech-at-bio.uio.no
I have a visitor who wants to use acridine orange (0.1%) in seawater (or artificial seawater) to stain nuclei/DNA in limpet eggs and sperm during fertilisation. It is done by other workers, but so far we've found no information as to how long it takes to stain things. Any suggestions? Any pH considerations?
Also, does it work on fresh (i.e. marine, seawater)and formalin-fixed material with equal efficiency.
Finally, we understand that the fluorescence is short-lived - any comments as to how long a time period would be helpful.
Many thanks
Keith Ryan Marine Biological Association & University of Plymouth UK
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ through the North Carolina State University Department of Continuing and Professional Education is now in its 19th year. The course will be presented October 30 - November 1, in Raleigh, NC. This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to
http://members.AOL.com/IPCourse/
or call Cindy Allen, 919-515-8171, at North Carolina State University Dept. of Continuing and Professional Education.
I am gathering data on enviornmental SEMs. Hopefully I'll be setting up a research facility within the coming months.
Does anyone have preference to systems?
What, if any differences are there for maintinence compared to a standard SEM?
COST? (ouch)
Thanks,
Tim Quinn University of Kansas Natural History Museum & Biodiversity Research Center 1345 Jayhawk Blvd 414A Lawrence, KS 66045 785-864-4556 tquinn-at-ku.edu
That *is* incredible. Chip has length of 4080 pixels by 15 micron pixel pitch = 61.2 mm. Area is then 37.45 square cm or 5.8 square inches. Plate (film) size is 3.25 inches by 4.00 inches = 13 square inches. Chip area is 45% of film area... -Mike
Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } This is incredible. I would suggest that this is a first EM digital camera } which produces comparable to the film image quality. The chip is huge, so } they have 80-90% area of the film on the chip. The images are very sharp. } The technical problem there was to read information from such huge } chip. They used new technology when chip (virtually?) splitted on quarters } and read in parallel. They did a great job. The camera is very expensive } by the way. } } Sergey } } At 11:00 AM 9/19/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I didn't get to the MSA meeting this summer, but I heard through the } } grapevine about Gatan's new UltraScan 1000 and saw their poster. I was very } } impressed with the 4K x 4k montage taken at 5k mag. It looked } } publishable.I'd be interested in hearing comments/impressions from anyone } } who a chance to use it. } } Thanks, } } } } Hank Adams } } Integrated Microscopy Core } } Molecular and Cellular Biology } } Baylor College of Medicine } } Houston, Tx 77030 } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu
The New York Society of Experimental Microscopists Announcing the 50th Anniversary Presidential Symposium
"FRONTIERS in MICROSCOPY"
Caspary Auditorium, Rockefeller University October 11, 2001 9:00 AM-4:00 PM Free and open to the Public
50 Years of NYSEM C. DeLemos-Chiarandini, New York University
George Palade, and the Early Days of Electron Microscopy in New York P. Satir, Albert Einstein College of Medicine
EM Tomography Reveals Cell Structure at 6nm Resolution J. R. McIntosh, University of Colorado
Multiphoton Imaging of the Molecular Dynamics of Life Processes W.Webb, Cornell University
Intravital Imaging of Tumor Cells J.Condeelis, Albert Einstein College of Medicine
Total Internal Reflectance Microscopy Visualizes Membrane Fusion Events S. Simon, Rockefeller University
Imaging Biochemistry Inside Cells P. Bastiaens, EMBL, Heidelberg
The Ribosome: Snapshots of a Molecular Machine in Motion J. Frank, HHMI, Wadsworth Center
Imaging RNA in Living Cells R. Singer, Albert Einstein College of Medicine
High Temporal and Spectral Resolution using Acousto-Optical Tunable Filters D. Farkas, Carnegie-Mellon University
**************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
You can culture your cells on plastic coverslips and embed them in resin. The embedded cells are good for both IFM and iEM but you have to work out the conditions. The coverslip is called Thermanox from EMS.
Greg
Gang Ning, M.D., Ph.D. Electron Microscopy Facility Medical College of Wisconsin
Andreas Brech wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I would appreciate any advice on the following question. I am planning to do } fluorescence microscopy and electron microscopy on the same sample. I would } like to do immunolabelling on cells grown in a culture dish followed by plastic } embedding of the same sample and further preparation for EM. } In order to find the right area of cells to be sectioned for EM (previously } detected by immuno fluorescence) I suppose one has to have some kind of grid } on the culture dish. I am aware of the fact that there are gridded coverslips } available, however, I always had a lot of trouble to remove the coverslip from } the glass after embedding (used lN2 and so on). So it would be very helpful if } anyone out there could provide any other working recipes, references for } methods or sources for culture dishes with some kind of grid on them. } } Dr. Andreas Brech } EM-Unit, Dept. of Biology, } University of Oslo. } P.O.Box 1062 Blindern, Room U 139 } Street address: Moltke Moes vei 32 } N-0316 Oslo 3 } Norway } Tel.: + 47-22 85 47 25/61 89 (work) } + 47-22 43 83 23 (privat) } Fax.: + 47-22 85 47 26 } e-mail.: abrech-at-bio.uio.no
Hi Listers, I have a question about a schottky tipped FE Microscope. Specifically, what is the best way to shut the beam off at the end of the day. We will be using probably everyday for long sessions, so would beam blanking be preferable to shutting off the extraction voltage. I am wondering about the effects of tip life, beam stability, and contamination. Any advice or past experience would be greatly appreciated. Thanks again Nick Aitken
Nicol Aitken Sample Preparation and Imaging Specialist Research and Development Semiconductor Insights Inc. email:nicol-at-semiconductor.com (613)599-6500 ext.4300
Yes, I did check your WEB-site in the beginning of my search a few month ago as well as many others. My initial criteria were the following:
-35 mm port mount; -price; -ability to have demo for the camera; -ability to have reasonable service in time.
You product was excluded from initial list for the following reasons:
- your Biocam camera, which was possible candidate is bottom-mounted;
- on your WEB-site at http://www.tvips.com/Contact_en.htm I did not find any information about your headquarter in US or contact phones, so I was not able to talk to your representative (I was using the telephone call as a first "try" of the customer service quality: if I was on hold for more than 5 min, I excluded the candidate from my list).
-location of your head office in Germany makes me skeptical about customer service quality company could provide in US.
I would like to mention that my criteria is subjective and based on my own way to make a business and spent my money. My decision is not related to the real quality of your products.
I would mention also that a few companies refused to give me demo on their cameras (or delayed with answer) and were excluded from the list as well.
Sergey.
At 02:30 PM 9/20/01 +0200, you wrote: } Dear Mr. Sergey Ryazantsev, } } I have read in your email, that you have studied cameras for TEM. } Have you studied our cameras as well? } } For example, this year we have had the following installations in California: } - Scripps in San Diego, F-224 at a CM-200FEG (Ron Milligan's lab) } - UC in San Diego, F-224/FastScan at a JEM-200EX (Mark Ellisman's lab) } - Quantum Dot Corporation in Hayward, TemCam-0124 at JEM-200CX } - Additionally just this month Bridget Carragher (together with her group) has } moved from Illinois to the Scripps Institute. She will mount another F-224 } at a } Tecnai-20FEG. } } If you want to know more about our US installations and our cameras, } please let } me know. } } Best wishes, } Ingo Daberkow } } BTW: We are in contact with Phoebe Stewart from the Department of Molecular & } Medical Pharmacology, UCLA School of Medicine } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Dr. Ingo Daberkow } Tietz Video and Image Processing Systems GmbH } Herbststrasse 7 } D-82131 Gauting, Germany } Tel: +49-89-8506567 } FAX: +49-89-8509488 } Internet: http://www.tvips.com/ } Email: ingo.daberkow-at-tvips.com } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hank, } } } } I think you wrong. They do have two new cameras 2x2K and 4x4K. This is } } real number of pixels on the chip. I also believe, Ultrascan1000 is much, } } much more expensive than you suggest. As per our conversation with Gatan } } people, they expect to start selling that cameras at the end of the year, } } so it's very unlikely the price is known for now (I did not ask them } } directly about the price, but have impression that it's much more expensive } } that previous models, which were in the $70K range). Current Gatan's } } cameras (600W/CW Multiscan for instance) already has capacity for multiple } } exposure I believe. Actually, it's not a properties of camera but } } software. It's called "Auto-montage" and you could "montage" as many } } pictures as you want. By the way, Multiscan cameras has 2-4x (not sure in } } exact number) bigger area of view than film, which sometime is very } } useful. Over last few month I was studying EM digital cameras on the } } makket very hard, so if you have questions, may be I could help. } } } } Sergey. } } } } At 02:42 PM 9/19/01 -0500, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Sara, actually the total package for the UltraScan 1000 is about } $100K. The } } } UltraScan 4000 is the one you've quoted and that is a 4kx4k camera. } The 1000 } } } has the 2kx2k camera and montages the field requiring 4 exposures. I } believe } } } that is the way it works. } } } } } } Hank } } } } } } -----Original Message----- } } } } From: Sara Miller [mailto:saram-at-duke.edu] } } } Sent: Wednesday, September 19, 2001 1:58 PM } } } To: Hank Adams } } } Subject: Re: Gatan's new UltraScan 1000 } } } } } } } } } Micrograph was lovely. Price tag was in range of $275 K--ouch! They } didn't } } } have one there on demo that I saw. } } } } } } Sara E. Miller, Ph. D. } } } P. O. Box 3712 } } } Duke University Medical Center } } } Durham, NC 27710 } } } Ph: 919 684-3452 } } } FAX: 919 684-3265 } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Steve Pfeiffer of Gatan just E.mail to me that price range for Ultrascan 1000 is about $100K. Which is much less than I expect to see for such camera.
Sergey
At 10:06 AM 9/20/01 -0500, you wrote: } Sergey, I just talked to Gatan about their UltraScan 1000 that tiles 4 } images together. The entire package with installation/training and the } Automontage plug-in minus the computer/monitor is about $100K. Using their } specs for the computer add another $4800 which includes a 21" monitor. The } CCD magnifies the image 1.3x-1.5x compared to film. } } Hank } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Wednesday, September 19, 2001 8:10 PM } To: Hank Adams; Microscopy-at-sparc5.microscopy.com } Subject: RE: Gatan's new UltraScan 1000 } } } Hank, } } I think you wrong. They do have two new cameras 2x2K and 4x4K. This is } real number of pixels on the chip. I also believe, Ultrascan1000 is much, } much more expensive than you suggest. As per our conversation with Gatan } people, they expect to start selling that cameras at the end of the year, } so it's very unlikely the price is known for now (I did not ask them } directly about the price, but have impression that it's much more expensive } that previous models, which were in the $70K range). Current Gatan's } cameras (600W/CW Multiscan for instance) already has capacity for multiple } exposure I believe. Actually, it's not a properties of camera but } software. It's called "Auto-montage" and you could "montage" as many } pictures as you want. By the way, Multiscan cameras has 2-4x (not sure in } exact number) bigger area of view than film, which sometime is very } useful. Over last few month I was studying EM digital cameras on the } makket very hard, so if you have questions, may be I could help. } } Sergey. } } At 02:42 PM 9/19/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Sara, actually the total package for the UltraScan 1000 is about $100K. The } } UltraScan 4000 is the one you've quoted and that is a 4kx4k camera. The } 1000 } } has the 2kx2k camera and montages the field requiring 4 exposures. I } believe } } that is the way it works. } } } } Hank } } } } -----Original Message----- } } } From: Sara Miller [mailto:saram-at-duke.edu] } } Sent: Wednesday, September 19, 2001 1:58 PM } } To: Hank Adams } } Subject: Re: Gatan's new UltraScan 1000 } } } } } } Micrograph was lovely. Price tag was in range of $275 K--ouch! They } didn't } } have one there on demo that I saw. } } } } Sara E. Miller, Ph. D. } } P. O. Box 3712 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-3265 } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
For the past 4 years we have run the thermal emitter in our Zeiss LEO DSM982 almost everyday Monday-Friday and off on weekends and vacations. The previous emitter (Zeiss OEM) lasted 14,900 hours over 3 years. The present emitter (from FEI) has 3,000 hours since November 2000. We routinely image at } 100kx with great results. What microscope do you have?
Jim
Nicol Aitken wrote:
Hi Listers, I have a question about a schottky tipped FE Microscope. Specifically, what is the best way to shut the beam off at the end of the day. We will be using probably everyday for long sessions, so would beam blanking be preferable to shutting off the extraction voltage. I am wondering about the effects of tip life, beam stability, and contamination. Any advice or past experience would be greatly appreciated. Thanks again Nick Aitken
Nicol Aitken Sample Preparation and Imaging Specialist Research and Development Semiconductor Insights Inc. email:nicol-at-semiconductor.com (613)599-6500 ext.4300
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
There may be a difference in TEM and SEM FEG use that I am not familiar with, I am primarily a TEM guy, however we keep our emission running as much as possible.
With our JEOL3000F FEGTEM we run the tip down maybe 4 times a year, other than that it is on and the valve between the gun and the column is closed when the instrument is not being used. There are several reasons for this.
It takes a couple of hours to run up and condition the HT at 300kV and then another couple of hours to run up the emission. This makes it impossible to switch on the emission for a day's use.
Having run up the emission it is not stable, the emission current keeps dropping. This exponential decrease takes several hours to stabilise (we allow 6 hours before we think it is really stable). Of course if you don't need a stable current (eg. for HREM imaging) you can still use it.
The most likely time for any damage to the tip (apart fom accidents) is when it is heated or cooled so we don't want to do that more than neccessary.
I guess that there is finite life of the tip so it would be pointless to keep it running for a long time if the instrument is not being used. Does anyone know why a tip should eventually fail if operating conditions are optimised?
Our present tip (the first) has been running during factory tests and since installation in Oxford in 1998 and has run for over 21,500 hours.
Regards, Ron ps. If our tip now fails I'm going to blame you guys for raising the subject.
} Nicol, } } For the past 4 years we have run the thermal emitter in our Zeiss LEO } DSM982 almost everyday Monday-Friday and off on weekends and vacations. The } previous emitter (Zeiss OEM) lasted 14,900 hours over 3 years. The present } emitter (from FEI) has 3,000 hours since November 2000. We routinely image } at } 100kx with great results. } What microscope do you have? } } Jim } } } Hi Listers, } I have a question about a schottky tipped FE Microscope. Specifically, what } is the best way to shut the beam off at the end of the day. We will be using } probably everyday for long sessions, so would beam blanking be preferable to } shutting off the extraction voltage. I am wondering about the effects of tip } life, beam stability, and contamination. Any advice or past experience would } be greatly appreciated. } Thanks again } Nick Aitken } } Nicol Aitken } Sample Preparation and Imaging Specialist } Research and Development } Semiconductor Insights Inc. } email:nicol-at-semiconductor.com } (613)599-6500 ext.4300 } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } Unit-2131 } Storrs, CT 06269-2131 } bsgphy3-at-uconnvm.uconn.edu } 860 486-2914 voice } 860 486-1936 fax } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
For the most complete information on fluorescence probes (which you may already know), try URL: http://www.probes.com/; Search: "Acridine Orange" for a plethora (my son accused me of a "plethora of harassment", the other night!) of information and citations.
You might also consult Hayat, MA, Stains and Cytochemical Methods, Plenum, 1993, ISBN: 0-306-44294-9 for an informative, though not explicit, description of AO and its applications and properties.
For the reason that AO has been used extensively in flow cytometry, you should also look into the several chapters that are relevant to your question in: Methods in Cell Biology, vol 33, Flow Cytometry, Darzynkiewicz and Crissman(Eds.)Academic Press, 1990, ISBN: 0-12-564133-8 or 0-12-203050-8.
pH is important to the fluorescent properties of the dissolved dye, but I have not observed such effects in my experience which has been confined to viewing virus-infected cultured cells (vital, but usually fixed/dead). My original histochemical use of AO fluorescence - early '60's - was to distinguish between DNA and RNA - until it was pointed out by virologists that there were some single-stranded DNA viruses that could interfere with interpretation. Now we know that AO can be used to intercalate preferentially in transcribing DNA, thus providing an indication of RNA production.
I hope this helps, and
Good luck,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Keith Ryan } Reply To: kpr-at-mba.ac.uk } Sent: Thursday, September 20, 2001 7:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM - acridine orange - help } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello Listers } } I have a visitor who wants to use acridine orange (0.1%) in seawater (or } artificial seawater) to stain nuclei/DNA in limpet eggs and sperm during } fertilisation. It is done by other workers, but so far we've found no } information as to how long it takes to stain things. Any suggestions? Any } pH considerations? } } Also, does it work on fresh (i.e. marine, seawater)and formalin-fixed } material with equal efficiency. } } Finally, we understand that the fluorescence is short-lived - any comments } as to how long a time period would be helpful. } } } Many thanks } } Keith Ryan } Marine Biological Association } & University of Plymouth } UK } } PLymouth } } }
my experience is the same as jim's. i've got about 10Khours on an fei emitter now...still works great at } 500KX. its "on" all week and "off" most weekends.
b-
} X-Comment: UConnVM.UConn.Edu: Mail was sent by d40h44.public.uconn.edu } Date: Thu, 20 Sep 2001 17:59:41 -0400 } To: Microscopy-at-sparc5.microscopy.com } From: Jim Romanow {bsgphy3-at-uconnvm.uconn.edu} } Subject: Thermal FE } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks for the fast reaction, *but* you have forwarded my direct email to you to the listserver as well.
Do you really want to discuss this matter at the listserver?
Of course, I am willing to discuss the advantage/disadvantage of our cameras in publicity, buy I fear this might be regarded as advertisement!?
Therefore, at first, I want to know the opinion of the list server community! Nestor, maybe do you can comment it?
Best wishes, Ingo
Sergey Ryazantsev wrote:
} Dear Ingo } } Yes, I did check your WEB-site in the beginning of my search a few month } ago as well as many others. My initial criteria were the following: } } -35 mm port mount; } -price; } -ability to have demo for the camera; } -ability to have reasonable service in time. } } You product was excluded from initial list for the following reasons: } } - your Biocam camera, which was possible candidate is bottom-mounted; } } - on your WEB-site at http://www.tvips.com/Contact_en.htm I did not find } any information about your headquarter in US or contact phones, so I was } not able to talk to your representative (I was using the telephone call as } a first "try" of the customer service quality: if I was on hold for more } than 5 min, I excluded the candidate from my list). } } -location of your head office in Germany makes me skeptical about customer } service quality company could provide in US. } } I would like to mention that my criteria is subjective and based on my own } way to make a business and spent my money. My decision is not related to } the real quality of your products. } } I would mention also that a few companies refused to give me demo on their } cameras (or delayed with answer) and were excluded from the list as well. } } Sergey. }
Only the FEI ESEM allows you to have water in the chamber. If I had the money I would get the FEGESEM.
Re maintainance, we do not have any extra problems with our XL30 ESEM as our wet bullet apertures are replaced at service. Our contract gives us 4 services a year. It would be useful to get comments from a user without a service contract.
Dave
On Thu, 20 Sep 2001 10:37:47 -0500 "Quinn, Tim Lee" {tquinn-at-ku.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listservers, } } I am gathering data on enviornmental SEMs. Hopefully I'll be setting up a } research facility within the coming months. } } Does anyone have preference to systems? } } What, if any differences are there for maintinence compared to a standard } SEM? } } COST? (ouch) } } Thanks, } } Tim Quinn } University of Kansas } Natural History Museum & } Biodiversity Research Center } 1345 Jayhawk Blvd 414A } Lawrence, KS 66045 } 785-864-4556 } tquinn-at-ku.edu }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I am interested in subscribing to the microscopy listserver.
I have a question about immunogold labeling. I am interested in surface labeling receptors and examining patterns of internalization. Has anyone tried to label the surface receptor with a primary antibody in the live cell, induce internalization, fix and embed the cells. Then thin section the preparation and do postembedding with a secondary antibody tagged with a gold particle? Do you know if the primary antibody will survive the embedding process with low-termperature embedding with lowicryl? Thanks for your help. Michelle Adams
Hi All, I hope to be installing a new TEM soon and unfortunately have some concerns about magnetic field interference with the column.
Being a new "immigrant" to the USA I am not up to speed with vendors on this side of the Atlantic. I need to know who supplies field canceling systems in the USA - calls from vendors welcome. I would also be interested in colleagues experiences with individual systems. I may have to deal with particularly high field strengths in the vicinity of the microscope so would be interested in what is the maximum field strength that such systems can compensate. Another alternative would be to look into Mu metal lining for the microscope room. Again info on vendors would be most useful as well as anyone's experience of using this method of shielding.
Many thanks
Chris
Christopher Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility Cell Biology Department University of Texas Southwestern Medical Center at Dallas Dallas, TX, 75390-9039 phone +1 214 648 2827 fax +1 214 648 6408 christopher.gilpin-at-utsouthwestern.edu
On a related note we just purchased a camera from Ingo. We have been extremely happy with the performance. The camera was installed about one mouth after tvips received the purchase order and the instating went very smoothly. If anyone has questions please contact me.
Glenn
} Sergey, } } Thanks for the fast reaction, *but* you have forwarded my direct email to } you to } the listserver as well. } } Do you really want to discuss this matter at the listserver? } } Of course, I am willing to discuss the advantage/disadvantage of our } cameras in } publicity, buy I fear this might be regarded as advertisement!? } } Therefore, at first, I want to know the opinion of the list server community! } Nestor, maybe do you can comment it? } } Best wishes, } Ingo } } } Sergey Ryazantsev wrote: } } } Dear Ingo } } } } Yes, I did check your WEB-site in the beginning of my search a few month } } ago as well as many others. My initial criteria were the following: } } } } -35 mm port mount; } } -price; } } -ability to have demo for the camera; } } -ability to have reasonable service in time. } } } } You product was excluded from initial list for the following reasons: } } } } - your Biocam camera, which was possible candidate is bottom-mounted; } } } } - on your WEB-site at http://www.tvips.com/Contact_en.htm I did not find } } any information about your headquarter in US or contact phones, so I was } } not able to talk to your representative (I was using the telephone call as } } a first "try" of the customer service quality: if I was on hold for more } } than 5 min, I excluded the candidate from my list). } } } } -location of your head office in Germany makes me skeptical about customer } } service quality company could provide in US. } } } } I would like to mention that my criteria is subjective and based on my own } } way to make a business and spent my money. My decision is not related to } } the real quality of your products. } } } } I would mention also that a few companies refused to give me demo on their } } cameras (or delayed with answer) and were excluded from the list as well. } } } } Sergey. } }
Glenn Fried Imaging Technology Group Beckman Institute University of Illinois phone 217 333 5493 Fax 217 244 6219
I shut extraction voltage off only when I was going on a long vacation and when I was told there could be a possibility of power outage (I put a microscope in a stand-by mode). Thermo cycling is the second worst thing you can do to the tip after power outage.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Nicol Aitken [mailto:nicol-at-semiconductor.com] } Sent: Thursday, September 20, 2001 3:09 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Thermal FE } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } Hi Listers, } I have a question about a schottky tipped FE Microscope. } Specifically, what } is the best way to shut the beam off at the end of the day. } We will be using } probably everyday for long sessions, so would beam blanking } be preferable to } shutting off the extraction voltage. I am wondering about the } effects of tip } life, beam stability, and contamination. Any advice or past } experience would } be greatly appreciated. } Thanks again } Nick Aitken } } Nicol Aitken } Sample Preparation and Imaging Specialist } Research and Development } Semiconductor Insights Inc. } email:nicol-at-semiconductor.com } (613)599-6500 ext.4300 } } }
I have been getting mail from root-at-mail.advancelink.com all afternoon. Twenty or more of these emails. Other (I think that are on this list) have been getting the same email but from my email address. I have not sent these messages.
I have this bad feeling that this is a virus and it I have gotten from the listserver somehow. Could the admin of this listserver check this out.
What's the rationale for turning it off over weekends?
My system uses, I believe, a Denka ZrO/W filament in my Amray 305FE gun assembly. I never shut the gun off. The main problem with this particular SEM is that the gun valve is manual. If the gun is on and the gun valve is closed, there is most likely to be an air burp into the gun chamber and will poison the filament. This process also typically will burn the valves O-rings due to the high intensity FE emitter beam. So the valve is never closed until after turning off the filament.
The only reason I have found to worry about closing the gun valve is if there is a power failure. That would shut down the filament and all pumps (mech, turbo, column and gun ion pumps). I solved this by using two Toshiba 1400XL+ 6KVA dual conversion UPS units. Now, I shut the console off but keep it leaves the pumps and gun running. Plus, the UPS units
I log gun and column vacuum every few days along with extractor current. Then, about every month, I measure gun brightness. Running the way I am now, brightness is +- no more than about 0.4nA. Iext may change 3-6uA but brightness is way less affected by changes in Iext. When Iext and brightness start to drop or waver, that is an onerous sign that the gun is starting to fail.
I would tend to stick with this pattern since I know that bringing up a new gun takes about a week for the Iext and brightness to stabilize. I suspect that there would be some proportional time to stabilize if shut down over a weekend or for some extended time. Since I don't use the SEM every day, if shutting off the filament during some periods of time would extend its life, that is very desirable.
I have 9,020 hours on this gun since newly installed. It is very stable.
gary g.
At 05:51 AM 9/21/2001, you wrote:
} hi- } } my experience is the same as jim's. i've got about 10Khours on an fei } emitter now...still works great at } 500KX. its "on" all week and "off" } most weekends. } } b- } } } } X-Comment: UConnVM.UConn.Edu: Mail was sent by d40h44.public.uconn.edu } } Date: Thu, 20 Sep 2001 17:59:41 -0400 } } To: Microscopy-at-sparc5.microscopy.com } } From: Jim Romanow {bsgphy3-at-uconnvm.uconn.edu} } } Subject: Thermal FE } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Nicol, } } } } For the past 4 years we have run the thermal emitter in our Zeiss LEO } } DSM982 almost everyday Monday-Friday and off on weekends and vacations. The } } previous emitter (Zeiss OEM) lasted 14,900 hours over 3 years. The present } } emitter (from FEI) has 3,000 hours since November 2000. We routinely image } } at } 100kx with great results. } } What microscope do you have? } } } } Jim } } } } } } } } } } } } } } } } Nicol Aitken wrote: } } } } Hi Listers, } } I have a question about a schottky tipped FE Microscope. Specifically, what } } is the best way to shut the beam off at the end of the day. We will be using } } probably everyday for long sessions, so would beam blanking be preferable to } } shutting off the extraction voltage. I am wondering about the effects of tip } } life, beam stability, and contamination. Any advice or past experience would } } be greatly appreciated. } } Thanks again } } Nick Aitken } } } } Nicol Aitken } } Sample Preparation and Imaging Specialist } } Research and Development } } Semiconductor Insights Inc. } } email:nicol-at-semiconductor.com } } (613)599-6500 ext.4300 } } } } James S. Romanow } } The University of Connecticut } } Physiology and Neurobiology Department } } Electron Microscopy Facility } } Unit-2131 } } Storrs, CT 06269-2131 } } bsgphy3-at-uconnvm.uconn.edu } } 860 486-2914 voice } } 860 486-1936 fax } } } } } } } } } } } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875
I received this reply from a person about the cron daemon problem this afternoon. If anyone has continuing problems, I suggest you contact them directly.
--John chandler-at-colostate.edu Colorado State University
} From: "Kim M. Shiu" {kmshiu-at-mail.advancelink.com} } Subject: Re: Random mail from your server to me } To: chandler-at-lamar.colostate.edu (John Chandler) } Date: Fri, 21 Sep 101 16:23:52 -0700 (PDT) } MIME-Version: 1.0 } Status: RO } } Hi, } } There was a problem with a program on our server. It was not intentional } to send multiple copies or spam anyone. The problem has been corrected } and we apologize for any inconvenience it caused. } } Thanks. } } Kim Shiu } Advancelink } } } } } I sent this message to postmaster-at-advancelink.com and the messages continue } } to be sent. Please attend to this immediately. } } } } --John } } } } FYI, I have receive 7 messages from your mail server this afternoon similar } } or identical to the one below. This might indicate a situation that } } requires your attention. } } } } John Chandler } } Colorado State University } } chandler-at-lamar.colostate.edu } } } } } } } } ==============================================================================
} } } } } } Received: from mail.advancelink.com ([207.214.173.11]) by } } lamar.ColoState.EDU (AIX4.3/8.9.3/8.8.8) with ESMTP id NAA237230 for } } {john.chandler-at-colostate.edu} ; Fri, 21 Sep 2001 13:28:56 -0600 } } Received: (from root-at-localhost) } } by mail.advancelink.com (8.8.5/8.8.5) id LAA29356; } } Fri, 21 Sep 2001 11:35:00 -0700 (PDT) } } Date: Fri, 21 Sep 2001 11:35:00 -0700 (PDT) } } Message-Id: {200109211835.LAA29356-at-mail.advancelink.com} } } From: root-at-mail.advancelink.com (Cron Daemon) } } To: coccust-at-mail.advancelink.com } } Subject: Cron {coccust-at-mail} csh -x ~/bin/get-dist } get-dist.log } } X-Cron-Env: {SHELL=/bin/sh} } } X-Cron-Env: {HOME=/usr/home/coc/coccust} } } X-Cron-Env: {LOGNAME=coccust} } } X-Cron-Env: {USER=coccust} } } Status: } } } } cd /usr/home/coc/coccust } } grep [a-z] /usr/home/coc/cocinfo/MAILING-LISTS/CURRENT-LIST.txt } } chown coccust .forward } } chown: Command not found. } } chown coc .forward } } chown: Command not found. } } chmod 664 .forward } } exit } } } } } } }
Greetings Stefan, My initial reaction is that it is not common to use murine (mouse) monoclonal antibodies on mouse tissue. The reason for this is that if these murine antibody hybridomas were derived from mouse B-cells, and these B-cells produced antibodies to a mouse protein, one would expect an autoimmune response in the original mouse. In other words, it not be logical to produce an antibody against mouse proteins in a mouse because either: 1.) a secondary immune response would not be observed, or 2.) the ensuing autoimmune response would kill or at least severly debilitate the mouse. I guess recombinant monoclonal murine antibodies against mouse proteins could be produced however. A more likely scenario is that these mouse monoclonal antibodies are against a protein antigen derived from another species. A couple of important controls are in order. The simplest is to assess non-specific binding of your secondary (gold-conjugate) antibody. To do this you might block your tissue as usual, apply PBS or murine pre-immune sera to the tissue in place of the primary antibody, and then apply your secondary antibody. Another important control is to asses non-specific binding of your primary antibody. This is an isotype control. You should be able to find out the isotype of your monoclonal antibody from the source (eg. IgG K2B). Then obtain a monoclonal antibody of the same isotype which is not specific for any mouse protein (eg. anti-dansyl or something). This is used in place of the mouse specific monoclonal to assess non-specific binding due to charge properties. An isotype control for the secondary antibody could be conducted as well to assess the presence of tissue internal antibodies. -Karl G. _________________________________________________ Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology Room B650J University of Illinois at Urbana-Champaign Urbana, IL 61801 Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
Greetings Listers, As someone new to the challenge of managing multiuser equipment, I have come to realize that it will be important to devise an effective strategy to minimize wear and tear on instrumentation. It will also be important to attempt to try to salvage the results of the inevetable mistakes that users make. I am hoping some of the seasoned veterans out there can offer some advice. My question concerns how best to try to clean non-oil objectives which have been used with oil. I was previously under the impression that attempting this was futile, however I am hoping that I'm wrong. A collegue of mine suggested sonicating in acetone for a couple of minutes, but I'm hoping a gentler approach will suffice. Would the use of a surfectant or detergent like NP-40 or tween or triton-X be a viable appoach? Are the cements used in objectives commonly water soluble or are there safe organic solvents to use to clean objectives? I also have some nice oil-immersion objectives with dried films of oil on them. Any suggestions? If any one has some good suggestions for general references pertaining to the cleaning and maintenence of objective lenses, I would welcome the correspondence. I'm looking into the matter presently, but a point in the right direction might expedite things. Thank you. -Karl Garsha _________________________________________________ Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology Room B650J University of Illinois at Urbana-Champaign Urbana, IL 61801 Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
I have been getting these messages, too, but they seem to have stopped now. I called advancelink, but only got a machine. My hunch is it is one of those outfits that swamp your inbox with junk email. I simply told me email application to just delete anything from advancelink.
As for a virus: It might be a virus on the advancelink servers, but the messages I got contained some Unix commands (like "cron" and "grep"). To me it looked more like some *really* clever person piped his Unix commands into some form of mailing application.
As far as I could tell, none of the mails had any attachments, and I don't think a mail application would run Unix commands embedded in an email.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Brian J Laughlin [mailto:brjlau18-at-US.ibm.com] Sent: Friday, September 21, 2001 3:03 PM To: Microscopy-at-sparc5.microscopy.com
I have been getting mail from root-at-mail.advancelink.com all afternoon. Twenty or more of these emails. Other (I think that are on this list) have been getting the same email but from my email address. I have not sent these messages.
I have this bad feeling that this is a virus and it I have gotten from the listserver somehow. Could the admin of this listserver check this out.
Whatever you do, please *don't* sonicate in acetone! This will dissolve the adhesives that seal the lens and keep the lens elements in place, and it will make the lens leaky. Acetone inside the lens will put you in deep trouble.
You can use anhydrous ether *very sparingly.* Use a cotton-tipped applicator or a "Q-Tip" moistened with just a little ether and work quickly, gently using circular motions on the front lens element. The ether will evaporate rapidly, and you don't want to rub the lens with a dry cotton applicator, so you will probably want to use several applicators. Ether is good for this purpose because it evaporates so quickly. It's not around long enough to damage the sealants and the adhesives.
You can follow this with a surfactant if necessary, but the ether should do the trick. Some surfactants can leave a film on the surface of the lens unless you work quickly and follow with a rinse of distilled water (using cotton applicators, *not* a stream from a squeeze bottle). I would avoid the use of surfactants if at all possible. If you do need some surfactant action, first try a little commercial or consumer glass cleaner. Check the labels for contents.
Remember, use ether sparingly, only enough to moisten the cotton applicator. Avoid flooding the front lens element with ether (or alcohol, or anything, for that matter).
Also, please note the usual cautions when working with ether (open flames, sparks, electrical discharges, etc. should be avoided, use adequate ventilation or work under a fume hood, etc.)
If you feel uncomfortable performing these steps, please call in a good microscope service technician who knows what he/she is doing. I'm sure there is one in your area. This can save you a lot of headaches (and a lot of lenses).
Good luck!
Bob Chiovetti GTI Microsystems Leica Exclusive Regional Dealer Desert Southwest (Arizona, New Mexico, West Texas USA)
***Disclaimer: These are my own opinions, gained from experience, not necessarily those of GTI Microsystems or Leica Microsystems.***
In a message dated 09/22/2001 9:34:08 AM US Mountain Standard Time, garsha-at-itg.uiuc.edu writes:
{ { Greetings Listers, As someone new to the challenge of managing multiuser equipment, I have come to realize that it will be important to devise an effective strategy to minimize wear and tear on instrumentation. It will also be important to attempt to try to salvage the results of the inevetable mistakes that users make. I am hoping some of the seasoned veterans out there can offer some advice. My question concerns how best to try to clean non-oil objectives which have been used with oil. I was previously under the impression that attempting this was futile, however I am hoping that I'm wrong. A collegue of mine suggested sonicating in acetone for a couple of minutes, but I'm hoping a gentler approach will suffice. Would the use of a surfectant or detergent like NP-40 or tween or triton-X be a viable appoach? Are the cements used in objectives commonly water soluble or are there safe organic solvents to use to clean objectives? I also have some nice oil-immersion objectives with dried films of oil on them. Any suggestions? If any one has some good suggestions for general references pertaining to the cleaning and maintenence of objective lenses, I would welcome the correspondence. I'm looking into the matter presently, but a point in the right direction might expedite things. Thank you. -Karl Garsha _________________________________________________ Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology Room B650J University of Illinois at Urbana-Champaign Urbana, IL 61801 Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu } }
JoAnn: It has been a very long time, but I have used lanthanum quite extensively in the past. And, yes, you do get a lot of precipitate (maybe flocculant is a better word) in the various solutions. One of the things I did was to make sure the washes were clean prior to starting the dehydration process. I have not found that this flocculant material adversely affects the tissue (or, for that matter, cultured cells). However, if there is a lot of remnant material, it can/will appear as large clumps in the EM. If you are going for cell surface labeling/coating, the large clumps can obscure any fine details. As with a number of other non-specific labeling methods, cleanliness, assuring adequate washing and attention to the minutiae will give you good results. Best of luck.
Roger Moretz, Ph.D. Dept. of Toxicology Boehringer Ingelheim Pharmaceuticals
On Mon, 17 Sep 2001 19:43:59 -0500, JoAnn Buchanan wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Hello folks. I am trying to fix and stain with lanthanum nitrate and | have followed procedures in Hyatt's book. It mentions that lanthanum | will precipitate in the cold, and when mixed with phosphate buffer. | I have used cacodylate buffer, room temperature solutions and checked | the pH of my buffer(7.2), but I am seeing a lot of ppt in all the | solutions (glut fix, wash and osmium fix). I am wondering if anyone | has any suggestions about how to alleviate this problem. Is the | precipitate harmful to the tissue? Can I filter the solutions to get | rid of big clumps? Is this normal? I would appreciate any feedback | from someone who has used the stuff. Thanks, JoAnn | | JoAnn Buchanan | Molecular and Cellular Physiology | Stanford University School of Medicine | Stanford, CA 94305 | 650-723-5856 |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
Does anyone know if it is possible to do TEM autoradiography of organic compounds labelled with 14C?
Bob
Robert R. Wise, Ph.D. Associate Professor of Plant Physiology Department of Biology and Microbiology University of Wisconsin Oshkosh Oshkosh, WI 54901 tele: (920) 424-3404 fax: (920) 424-1101 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
Bob: There is a book called "Techniques of Autoradiography" by AW Rogers (1979, Elsevier) that is a gold mine of info about LM & EM autoradiography. He states that 14C quotes Salpeter & Salpeter, 1971 as showing a C14 specimen giving a HD (half diameter - distance of an exposed silver grain from its source) resolution of 180 - 285 nm. of course, a problem with lots of organic compounds is to fix them in place during the various prep steps. good luck. tom
} } } Does anyone know if it is possible to do TEM autoradiography of organic } compounds labelled with 14C? } } Bob } } Robert R. Wise, Ph.D. } Associate Professor of Plant Physiology } Department of Biology and Microbiology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } tele: (920) 424-3404 } fax: (920) 424-1101 } wise-at-uwosh.edu } http://www.uwosh.edu/departments/biology/wise/wise.html
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Energy-Loss Spectroscopy of Polymers and Biomaterials at Stevens Institute of Technology
There is a post-doctoral opportunity within the Department of Chemical, Biochemical, and Materials Engineering at the Stevens Institute of Technology to develop and apply techniques associated with spatially-resolved electron energy-loss spectroscopy. Application will be made to study the morphology in biological tissue, in synthetic polymers, and in mixtures of these two. Much of this work will be done in collaboration with Unilever Research to pursue ongoing projects of mutual interest.
The ideal candidate will have experience in electron optics and electron
energy-loss spectroscopy and preferably in techniques of ultramicrotomy and cryo-ultramicrotomy.
The position can be filled as early as November, 2001. The appointment will be for one year with a renewal dependent upon performance and availability of funds. A multiyear appointment is anticipated.
The Stevens Institute of Technology is a small private university concentrated on engineering, science, and technology. The Stevens electron-optics laboratory contains a Philips CM20 FEG TEM/STEM, a Philips CM30 SuperTwin TEM, and a Leo 982 FEG SEM. The CM20 FEG TEM/STEM
is equipped with a Gatan Enfina ccd PEELS system and a Gatan Multiscan digital camera. Both are interfaced to an Emispec Vision acquisition and control system. The facility is fully equipped with cryomicrotomy and cryo-transfer capabilities to deal with frozen hydrated materials.
For further information please contact:
Professor Matthew R. Libera Dept. Chemical, Biochemical, and Materials Engineering Stevens Institute of Technology Hoboken, New Jersey 07030 ph: 201-216-5259 fax: 201-216-8306 mlibera-at-stevens-tech.edu
What about the surface of the Si(Li) detector to be installed on a FE-SEM ?
I was adviced to take a 30 square mm insted of the classical 10 square mm, to be installed on a cold FE-SEM. But if I look on my common analytical conditions on a classical W filament SEM, I work with current of 0.4 to 1 nA (mesured with a Faraday cup), with suffisant counting rates (1500 to 2500c/s, with 12 microsecond tc). All FE-SEM can give such current. Is it really necessary to take a detector with more surface (and probably less resolution) ?
I am interrested in any advice from users.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ae3-at-njit.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, September 24, 2001 at 01:42:00 ---------------------------------------------------------------------------
Email: ae3-at-njit.edu Name: Alexandre Ermoline
Organization: New Jersey Institute of Technology
Education: Graduate College
Location: Newark, NJ, USA
Question: I'd like to get nanoparticles on a grid to study them by TEM. I chose to use copper grid with plain carbon support film. But I doubt if the contrast introduced by the support may be negligible compared to that of the specimen because the thickness of the support film is 20-30 nm and the nanoparticles sizes are in the range 10-50 nm. Thank you for your kind attention Alex
Morning Bob, Two suggestions beyond Tom's good one: 1. "Thymidine Metabolism and Cell Kinetics", Elsevier, 1967. 2. "Autoradiography", Elsevier, Audrey Glauert's series in EM, softcovers. 14C and 3H have been used together for double autoradiography, because the penetrance of the beta particles from 14C is ~10X that of 3H, although both tend to average out at lower levels in practical situations. Some science on the subject: http://laxmi.nuc.ucla.edu:8248/M248_98/autorad/lm_interp.html. The problem with 14C IS the greater mean range and greater mean energy of the beta particles. This is one reason that when there is a choice, 3H label is preferred. Working out the bugs for autoradiography can take a long time and requires patience and attention to detail. I would recommend consideration of the following: Mol Cell Biochem 1995 Nov 22;152(2):167-73, "Hyperplasia in the rabbit bladder urothelium following partial outlet obstruction. Autoradiographic evidence," Monson FC, Sun L, Wein AJ, Levin RM.; for a LM method by which procedures for EM development might be expedited. If you are interested and would like more information, please ask.
Also, good luck,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: wise } Sent: Sunday, September 23, 2001 3:29 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Autoradiography of 14C? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all, } } Does anyone know if it is possible to do TEM autoradiography of } organic } compounds labelled with 14C? } } Bob } } Robert R. Wise, Ph.D. } Associate Professor of Plant Physiology } Department of Biology and Microbiology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } tele: (920) 424-3404 } fax: (920) 424-1101 } wise-at-uwosh.edu } http://www.uwosh.edu/departments/biology/wise/wise.html } } } }
Morning Karl, My opinion only! Since many/most of our best objectives/oculars are coated for color correction, Please don't use acetone on an objective - EVER!!! Please don't use 100% ethanol, xylene or toluene on an objective - ever!? You may concoct your own cleaner with non-ionic detergent (25%) and small amounts of alcohol(s) and (ONLY IF REQUIRED) small - added when required - amounts of xylene or toluene. I have known optical technicians who would rather scrape a lens with the edge of a new scalpel blade or a fragment/piece of double-edged razor blade than expose a lens to any solvent for more than a brief moment for final - residual oil - removal. There are some who never "rub". They "blow" with compressed air! I have in my possession a product called "ROR" (URL: http://www.ror.net/ [NO RELATION TO COMPANY]) which smells like a regular glass cleaner, but has received "KUDOS" from photographic folks for its effect on residual oils on lenses. Thus, it appears that this is a product that does not remove color corrective coatings from photographic lenses. I have also had in my possession for many years a product I acquired from Xerox called: "Lens and Platen Cleaner", Reorder No. 8R1025 (6 bottles) and "Cleaning Absorbent" [really clean cotton], Reorder No. 8R25. I have used this for residual oil removal for much longer than ROR, and I have never been disappointed, though, of course, the Xerox cleaner was intended for the windows and optics in their copiers. The biggest problems in cleaning microscope optics are these: 1. corrective coatings which wear the more they are cleaned, 2. mixing oils from different manufacturers (mentioned in a recent email to the listserver), because they can react badly with one another and harden, 3. front surface mirrors which are usually inside fluorescent cubes or other complicated optical devices (these should be cleaned only by someone who can afford to replace them), 4. permitting recidivist users to continue using better microscopes, and 5. assuming that a new faculty member or post doc has been properly taught or has properly learned while a graduate student. (I will never forget the senior graduate student who removed an oil immersion lens on a brand new microscope and poured it full of oil (from the nearby pint bottle) as he followed the printed directions for its use!) I have NEVER kept a pint bottle near a microscope since!
To your specific, other, and possibly implied questions (more than you ever hoped for in one man's opinion): 1. Yes, you can clean the 'dry' objectives of caked on oil with fluids. These are easier than many older oil immersion objectives, because their exposed lens surfaces are usually flat. 2. Cleaning is preferable to discarding without trying. 3. Serious caked on oil can be scraped off (GENTLY!!!) before trying to finish cleaning with a solution. 4. No, please do not soak the lens in anything. 5. Please, do not subject a complicated multi-element lens system to immersion and ultrasound. You will NOT 'see the baby' by doing this. The ultrasound will actually tend to separate cemented lens combinations and to dissolve or fragment the cement that seals the face element to the surrounding metal jacket. NO, please, definitely NO ultrasound for microscope objectives, oculars, mirrors, filters, or prisms. I HAVE used ultrasound to help clean grease from microscope focusing mechanisms and from sticky leaf diaphragms, but after you dismantle your first leaf diaphragm, you will know what real trouble is. One way to help users is to predefine iris and field diaphragm adjustments for each objective and mark the positions of those adjustments so that indices are easily repeated. This is far more important for the iris diaphragm than the field, because most unschooled microscopists learn in their first class to use the iris diaphragm for contrast enhancement (a.k.a., resolution diminution!). By placing the settings in plain view you will give yourself the opportunity to review the purposes of the two diaphragms with each new user and prevent unnecessary exposure of microscope internals. Is it obvious that I have a fixation on these matters?
} ---------- } From: Karl Garsha } Sent: Saturday, September 22, 2001 12:32 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: fouled objectives } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings Listers, } As someone new to the challenge of managing multiuser equipment, I } have } come to realize that it will be important to devise an effective strategy } to } minimize wear and tear on instrumentation. It will also be important to } attempt to try to salvage the results of the inevetable mistakes that } users } make. } I am hoping some of the seasoned veterans out there can offer some } advice. My question concerns how best to try to clean non-oil objectives } which have been used with oil. I was previously under the impression that } attempting this was futile, however I am hoping that I'm wrong. A } collegue } of mine suggested sonicating in acetone for a couple of minutes, but I'm } hoping a gentler approach will suffice. Would the use of a surfectant or } detergent like NP-40 or tween or triton-X be a viable appoach? Are the } cements used in objectives commonly water soluble or are there safe } organic } solvents to use to clean objectives? } I also have some nice oil-immersion objectives with dried films of oil } on them. Any suggestions? } If any one has some good suggestions for general references pertaining } to the cleaning and maintenence of objective lenses, I would welcome the } correspondence. I'm looking into the matter presently, but a point in the } right direction might expedite things. } Thank you. } -Karl Garsha } _________________________________________________ } Karl Garsha } Light Microscopy Specialist } Imaging Technology Group } Beckman Institute for Advanced Science and Technology } Room B650J } University of Illinois at Urbana-Champaign } Urbana, IL 61801 } Tel: (217) 244-6292 } Fax: (217) 244-6219 } www.itg.uiuc.edu } } }
The October meeting of NESM (New England Society for Microscopy) will be held Tuesday, October 16, 2001 at JEOL USA, Inc. in Peabody, MA.
This meeting will take an unusual format. It will be dedicated to a single subject, electron back-scatter diffraction pattern analysis in the SEM and will rely entirely on our sustaining members for content. EBSD is a relatively new electron beam technique which allows the determination of the crystallography of solid samples on a microscopic scale. It can be applied to phase determination in multi-phase samples, or orientation determination in single-phase polycrystalline samples, the latter being especially useful if some form of texture is present, as in drawn wires or thin films, for example.
Four vendors of commercial EBSD systems will be part of our symposium exploring the technique. The first part of the meeting will have demonstrations of EBSD Systems by John Sutliff of HKL Technology, Inc. and Neil Rowlands and Richard McLaughlin of Oxfor Instruments Microanalysis Division.
The second half of the evening will have technical presentations on the Theory and Applications of EBSD by Stuart Wright of TSL/EDAX and Patrick Camus of Thermo-Noran, Inc.
There will be a buffet supper, courtesy of JEOL.
Members are invited to bring posters on the topic of EBSD analysis for display at the meeting. Please call Tony Garratt-Reed (617-253-4622) for more information if you are interested.
The pre-registration deadline is Friday, October 12th. Please contact Mary McCann at (617-484-7865) or by email: mccanns-at-tiac.net.
I refer you to NESM's website for more information (times, directions, registration fees, etc.) listed in the current newsletter (September 2001).
Peggy Sherwood, Corresponding Secretary NESM -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
I do much EDS with a tungsten filament. My experience is that my image resolution suffers when I try to collect images at the same conditions used for EDS. I think I am also normally in the 1 nA range.
Therefore, I would probably welcome the improved count rate/beam current ratio that would be possible with a 30 mm2 detector. I could probably accept a small decrease in peak resolution if I could more closely match the conditions for imaging and x-ray analysis. Of course, I would not want to pay too much for the improvement. How much more does a 30mm2 cost compared to a 10 mm2 detector?
Warren
At 08:48 AM 9/24/2001 +0200, you wrote:
} Hello all } } What about the surface of the Si(Li) detector to be installed on a FE-SEM } ? } } I was adviced to take a 30 square mm insted of the classical 10 square } mm, to be installed on a cold FE-SEM. But if I look on my common } analytical conditions on a classical W filament SEM, I work with current } of 0.4 to 1 nA (mesured with a Faraday cup), with suffisant counting } rates (1500 to 2500c/s, with 12 microsecond tc). All FE-SEM can give such } current. Is it really necessary to take a detector with more surface (and } probably less resolution) ? } } I am interrested in any advice from users. } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess } 67037 Strasbourg CEDEX } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
Hi Karl, I manage a multi-user optical microscopy facility. Brace yourself. You will have to resign yourself to the fact that no matter how carefully you explain things to your users, someone is going to drag a dry lens through oil, get fingerprints on the ocular lenses, and do things you hadn't imagined. I try (not always successfully) to keep a schedule of visually inspecting the lenses, etc and correcting the damage before it gets too far gone. both of our microscopes are inverted, so I have had to devise little "diapers" for the lenses to absorb the excessive amount of immersion oil my users feel they need, so that it doesn't drip down and work its way into the lenses or even the body of the 'scope itself! (yes, they are that sloppy). We routinely use a solution of Windex (the standard blue stuff)& water (1:1) to keep our lenses clean. It works very well on oil that is still liquid. For the encrusted lenses, you may need something stronger. I would call your sales/service rep for the microscope and ask what they recommend for their lenses, since the coating on the lenses may be damaged by different solvents. Vigilance pays. Good luck.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Does anyone know of a commercially available anti-GFP antibody conjugated to gold? (preferably 10 nm). I know we can use a 'bridging' step to get to the gold, but I wanted to know if there is a direct secondary available. Thanks in advance for your help.
Mary
Mary McKee MGH Renal Unit Charlestown, MA 02129 (617)726-3696
} Question: I'd like to get nanoparticles on a grid to study them by TEM. } I chose to use copper grid with plain carbon support film. But I } doubt if the contrast introduced by the support may be negligible } compared to that of the specimen because the thickness of the support } film is 20-30 nm and the nanoparticles sizes are in the range 10-50 } nm.
I am imaging Si nanoparticles of as small as 4 nm on carbon support film. There is really very little contrast between the Si and C, so I hope you're looking at something a bit more electron dense! I make my own carbon films, and I have no idea how thick they are. I'm using a LEO 912 EFTEM at 100kV, and I can easily see particles of 10 nm and, with a lot of fiddling around, down to just about 4 nm. Smaller than that and I am seeing what may be evaporated carbon particles.
I think the main trick is to have as thin a support film as you can get - I make my own with variable results, and Ted Pella has some ultrathin carbon on lacy Formvar, I think. Make sure your scope is well aligned. Use a high enough accelerating voltage that you can resolve the things, but not so high that you shoot right through them and don't get enough electrons scattered away to see them. And I hope you're using a heavier element than I am!
Good luck.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We typically examine SWCNT's by ultrasonically dispersing a small quantity in a 10 ml beaker of ethanol, remove the beaker from the ultrasonic bath, dip a holey carbon film supported on a copper grid into the solution to pick up some of the nanotubes, and then place the grid on bibulous paper to wick away the excess solvent. This sample can then be used for both TEM as well as SEM.
I would be careful not to put a large quantity of loose SWCNT's into your SEM because typically the commercially available carbon nanotubes are produced with magnetic catalysts (iron). The magnetic field of the final lens can pull them off your mount and into your lens! We had a researcher looking at nanotubes grown on a clump steel wool, part of the sample was pulled into the objective lens of our FE-SEM, we noticed a glow coming from the final lens with our chamber view camera when the infrared diodes were turned off! An expensive light bulb! The service engineer had to clean the final lens.
At 7:55 AM -0500 9/17/2001, Haeseong Lee wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ?? } Hello. } } Could anybody recommend the good method of sample preparation for } SEM imaging on carbon nano tubes? } The sample is a powder type including single wall carbon nanotubes (SWCNTs). } Thank you very much in advance. } } Haeseong Lee } } {mailto:haeseong-at-hanyang.ac.kr} haeseong-at-hanyang.ac.kr
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
Does anyone know of any documented EDS calibration (energy display, quantification, etc.) procedures that have been certified in any way? i.e. NIST, ISO-9000 or any other.
I have previously posted a request for information on 3D reconstruction from serial sections.....but I only recieved replies from vendors and people wanting a summary of the replies.
{ { {I was wondering what computer programs people are using for 3-D reconstruction of TEM serial sections. I am interested in peoples opinions of the software (i.e. ease of use, learning curve, cross-platform functionality, cost, etc.). Feel free to email me off list if you choose. Also vendors feel free to send me any info you might have on the products you sell.} } }
I would really like to hear from the users of these programs! I would be glad to submit a summary to all interested parties (yes I still have your addresses).
Thanks to all,
-EK
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
see our web page at http://www.mayo.edu/mcniven_lab/
Moran Scientific is running EDS and XRM Microanalysis Courses from Monday 15th October to Friday 19th October at the Old Bungonia School. These are Courses designed for Beginners and those more Advanced. This is our last call for any people interested in attending. Please email us at the earliest opportunity for more information. Kind Regards, Diana Moran Moran Scientific Pty Ltd P.O. Box 651 Goulburn NSW 2580 Australia Tel 02 4844 4234 (International, 61 2 48444234) Fax 02 4844 4291 (International, 61 2 48444291) Email {kmoran-at-goulburn.net.au} Web Page http://www.goulburn.net.au/~kmoran/ "Patience accomplishes its object, while hurry speeds to its ruin. Gulistan 1258"
Contrast depends on the TEM technique that you plan to use. If you need particle size and morphology, diffraction contrast (bright / dark field imaging) may be acceptable on the carbon support film. If it is lattice structure by phase contrast microscopy, holey carbon film coated grids may be a better option. In the latter case, Jose Yacaman, Materials Trans JIM, 40 (1999), p 141 - 145 may be of interest to you.
Best wishes.
On Monday, September 24, 2001 5:38 PM, ae3-at-njit.edu [SMTP:ae3-at-njit.edu] wrote: } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ae3-at-njit.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, } September 24, 2001 at 01:42:00 } ---------------------------------------------------------------------- } ----- } } Email: ae3-at-njit.edu } Name: Alexandre Ermoline } } Organization: New Jersey Institute of Technology } } Education: Graduate College } } Location: Newark, NJ, USA } } Question: I'd like to get nanoparticles on a grid to study them by } TEM. } I chose to use copper grid with plain carbon support film. But I } doubt if the contrast introduced by the support may be negligible } compared to that of the specimen because the thickness of the support } } film is 20-30 nm and the nanoparticles sizes are in the range 10-50 } nm. } Thank you for your kind attention } Alex } } ---------------------------------------------------------------------- } -----
we currentlly use low currents (0.80 nA) for oxide glass EDX analysis on a tungsten filament SEM. The count rate is approx. 2000 cps at 30% dead time using a process time which correponds to 129 eV resolution on Mn. the SiLi detector surface is 10 mm². For elements in the range 0.5 - 1% a higher current is necessary. For elements { 1% we use WDX if the % has to be known precisely.
I think that a 30 mm² would be better if you need to analyse oxides or organic compounds at very low SEM currents ( { 0.5 nA). (The distance detector - sample has to be considered, too). It is more convenient to use the same SEM currents for imaging and analysis. A lower resolution is not necessairely a bad choice, provided you record your own standards and use an FLS-deconvolution program.
kindly,
Jean Louis HEITZ
CRITT Matériaux-LNE Est 19 rue de Saint Junien, BP 23 F - 67305 SCHILTIGHEIM CEDEX tel. +33(0) 388 191 510 fax +33(0) 388 191 514 email jl.heitz-at-critt.fr web www.critt.fr
Hi there Does anyone know or can suggest how I can fix and either wax or resin embed sugarcane stem tissue for the purpose of in situ work. Have tried a number of protocols but the end result is almost always the same: tears in the sections, thus making it difficult to continue with the rest of the hybridization procedure. Any help or suggestion would suffice. Thanx Rakeshnie Ramoutar Msc Student Institute for Plant Biotechnology University of Stellenbosch
the ISO TC (technical comitee) 202 works on microanalysis (SEM EDX/WDX - EPMA). you can contact your local standardization agency for information.
best regards,
Jean Louis HEITZ
CRITT Matériaux LNE Est 19, rue de Saint Junien - BP 23 F - 67305 SCHILTIGHEIM CEDEX tel. +33(0) 388 191 510 fax +33(0) 388 191 514 email jl.heitz-at-critt.fr web www.critt.fr
I have the problem to visualize the cytoskeleton of cartilage cells (chondrocytes) in articular cartilage of wild type and knockout mice. They are suspected to show differences in cytoskeletal organization. What is the most appropriate way to obtain highest possible contrast for the cytoskeleton (tricks during fixation, embedding, poststaining etc?)? With many thanks in advance for your help
Matthias Mörgelin, PhD Dept of Cell and Moleculer Biology Lund University BMC, C12 S-221 84 Lund Sweden
Our multi-user facility is in the process of upgrading one of our microscope (fully motorized Zeiss) / camera (Photometrics) systems and we are also considering the purchase of the SmartCapture2 software from Digital Scientific (UK). However, there is no demo version of this software available and hence the (dis)advantages of the software ar difficult to judge. In order to be able to make a well founded decision it is of great importance to us to get information from existing users. We would be very glad if you could comment on the following, taking into account that the software has to be used in a multi-user environment
* SmartCapture2 in general * the ease of use (interface) * the flexibility of the software and more specifically * the autofocus functioning * the batch capture possibility * any other aspect which you find important
I would greatly appreciate your comments, both positive and negative, and I already thank you in advance for your time taken. Those willing can of course also respond ofline.
Lauran Oomen Manager Digital Microscopy Facility The Netherlands Cancer Institute Amsterdam The Netherlands tel. +31-205121889 oomen-at-nki.nl
Hello Eugene, There is an NIH derivative that I've used for various stereology applications called Object Image (http://simon.bio.uva.nl/object-image.html). It also has a module for creating wire frame outlines of serial section type of information. Once made, it even has the ability to export the wire frame data to another program that can rotate the 3D object. I like to use this program to collect statistical info on size and shapes of particles, and I've been pleased with its useability, and easy learning curve. I haven't done any 3D work with it, but I've played around with the included tutorial models - pretty fun. The software is mac-based, has modest processing requirements, and is free. Its a great software application to have in your "toolbox". Good luck.
-Brad
---------- From: Krueger, Eugene W. Sent: Monday, September 24, 2001 1:53 PM To: 'Microscopy-at-sparc5.microscopy.com' Subject: 3D reconstruction from serial sections, 2nd request
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Hello to all in the microscopy community,
I have previously posted a request for information on 3D reconstruction from serial sections.....but I only recieved replies from vendors and people wanting a summary of the replies.
{ { {I was wondering what computer programs people are using for 3-D reconstruction of TEM serial sections. I am interested in peoples opinions of the software (i.e. ease of use, learning curve, cross-platform functionality, cost, etc.). Feel free to email me off list if you choose. Also vendors feel free to send me any info you might have on the products you sell.} } }
I would really like to hear from the users of these programs! I would be glad to submit a summary to all interested parties (yes I still have your addresses).
Thanks to all,
-EK
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
see our web page at http://www.mayo.edu/mcniven_lab/
UOP LLC, located in northwest suburban Chicago, is an international leader in technology, products and services for the petroleum processing and separation industries. Our R & D Deptartment, which supports fundamental and market-focused R & D, has an immediate need for an Electron Microscopist to work with chemists and engineers throughout the company on structural and analytical characterization issues relating to catalytic, adsorbent, ion exchanger and metallurgy materials.
Technical assignments will include:
- Interaction and collaboration with solid state and inorganic chemists, applications specialists, and technical service personnel in applying appropriate microscopic and data analysis techniques to problem solving in catalytic and adsorbent materials fields. - Interaction and collaboration with existing microscopy group to effectively utilize SEM and EDXS facilities, including training of junior technical staff. - Development and continuation of in-house expertise in current and emerging scanning electron microscopy techniques and applications.
Minimum qualifications include:
- BS/MS with multi-year relevant work experience, or recent PhD in Geology, Chemistry, Materials Science, or Physics (preferred coursework in electron microscopy, solid state chemistry and physics). - Expertise in SEM and electron probe X-ray microanalysis techniques. - Knowledge of sample preparation techniques, including ultramicrotomy. - Excellent team and communication skills.
Knowledge of other microscopic characterization techniques, for example TEM and STEM, and industrial R&D experience desirable but not essential. Above all, we are seeking a motivated individual, who will demonstrate a willingness to learn and openness to acquiring new areas of expertise in characterization, chemistry and materials properties.
We offer a state-of-the-art scientific research environment coupled with exciting career opportunities coupled with a highly competitive compensation and benefits package including medical and dental coverage, 401(k) Plan, Pension Plan and tuition reimbursement.
US employment authorization required.
UOP is an Equal Opportunity Employer
Visit us at www.uop.com
---------------------------------------------------------------------------- ---- Additional Information Position Type: Full Time, Employee Ref Code: MB-01-11
---------------------------------------------------------------------------- ---- Contact Information UOP Internet Resume Manager cro-at-uop.com UOP LLC 25 E. Algonquin Road, P.O. Box 5017 Des Plaines IL 60017-5017 Fax: 847.391-2921
Amenex is giving up its laboratory space, so much of our equipment is for sale. Check out the eBay seller list for amenex-at-amenex.com, which includes:
ETEC Autoprobe scanning electron microprobe ETEC evaporative coater Wilson Tukon microhardness tester Bausch & Lomb Balphot metallograph Lasico digital filar eyepiece Philtec sectioner Buehler dual-motor polishing table Oscar Fisher print dryer
I can be reached at amenex-at-amenex.com or at (610) 430-0117 if you have any questions.
Best regards, George Langford amenex-at-amenex.com http://www.amenex.com/
I need the user's manual for an American Optical 880 series microtome. These microtomes clamp to a bench. The specimen is fixed and the knife is moved by a parallelogram mechanism.
Will glady defray photocopying and mailing expense.
We need to purchase a new water recirculator for our electron microscope (Philips CM-10). Is there a consensus of opinion on what the best recirculator is, reliability being the highest consideration? Thanks.
I have an unused LaB6 filament, made by FEI and dated 10-24-95, for an ISI SEM. It's been stored in a desiccator since it was purchased. The 'scope it was intended for is long gone. I will be pleased to send it to the first person who might be able to use it. Please contact me offline.
Thanks
Scott Robinson
Microscopy Suite, room B650J Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana IL 61801 217 265-5071 (office); 217 244-6219 (fax) sjrobin-at-uiuc.edu
POSTDOCTORAL POSITION AT THE NAVAL RESEARCH LABORATORY
The U.S. Naval Research Laboratory has an opening for a postdoctoral fellow with experience in transmission electron microscopy and a strong background in physical metallurgy and/or materials science. The successful candidate will study phase transformations and their resultant microstructures in materials of interest to the U.S. Navy. Current research efforts are focussed on steels, aluminum-based alloys, magneto-electronic thin films, multilayers, and various welding processes (friction stir, laser, gas metal arc). Selection of a specific program will depend on the programs available and interests of the candidate. This position will employ a variety of analytical equipment as needed. Microscopy facilities available include a Philips CM-30 TEM, a Hitachi H-9000 HRTEM, a Leo 1550 SEM with OIM & EDS, and a new JEOL 2100-FEF energy filtered TEM (to be installed next summer). Additional facilities include a FIB, a quenching and deformation dilatometer, a Gleeble thermo-mechanical simulator, a state-of-the-art isothermal heat treatment facility, and the NRL visualization laboratory for three-dimensional analysis of microstructures. If interested, please send resume and names of references to Richard Fonda, Code 6324, Naval Research Laboratory, Washington, DC 20375. If you have any questions regarding this position, please contact either George Spanos (202-767-5799, spanos-at-nrl.navy.mil) or Richard Fonda (202-767-2622, fonda-at-nrl.navy.mil) for further details. NOTE: this position is only open to US citizens or residents with a green card. ________________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 ________________________________________________________________
I have looked at particles down to 1.5 nm in diameter, using grids coated with incredibly thin carbon film (1-2 nm thick). I get very high particle contrast, and the particles I look at aren't very dense either. I purchase these from the University of Chicago. The contact person is Dr. Robert Joseph (BOB-at-befvax.uchicago.edu). I am sure he will be more than delighted to sell you some.
Regards, Thearith
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Monday, September 24, 2001 11:55 AM To: ae3-at-njit.edu Cc: Microscopy-at-sparc5.microscopy.com
Hi, Alex-
} Question: I'd like to get nanoparticles on a grid to study them by TEM. } I chose to use copper grid with plain carbon support film. But I } doubt if the contrast introduced by the support may be negligible } compared to that of the specimen because the thickness of the support } film is 20-30 nm and the nanoparticles sizes are in the range 10-50 } nm.
I am imaging Si nanoparticles of as small as 4 nm on carbon support film. There is really very little contrast between the Si and C, so I hope you're looking at something a bit more electron dense! I make my own carbon films, and I have no idea how thick they are. I'm using a LEO 912 EFTEM at 100kV, and I can easily see particles of 10 nm and, with a lot of fiddling around, down to just about 4 nm. Smaller than that and I am seeing what may be evaporated carbon particles.
I think the main trick is to have as thin a support film as you can get - I make my own with variable results, and Ted Pella has some ultrathin carbon on lacy Formvar, I think. Make sure your scope is well aligned. Use a high enough accelerating voltage that you can resolve the things, but not so high that you shoot right through them and don't get enough electrons scattered away to see them. And I hope you're using a heavier element than I am!
Good luck.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu *
* Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf*
our group is involved in developing a program for 3-d reconstruction of TEM serial sections for modeling processes in prokaryotic cells. I asked the person responsible for the images stacking part (not a microscopist), and will try to translate his answer: It depends on what someone would want to do with the reconstruction. If you wanted to just look at the image of the reconstruction, then there is some software that you can kind of do this (with a lot of fiddling of the stack of data to get it into the program). The two packages are IBM's OpenDX and the VTK toolkit. If the purpose is to produce 3-D modeling geometry (like what we want to do), then there is nothing available to his knowledge. He says this is partly because there is no 3-D modeling code available that could consume such data. I do know of two groups that are doing section-by-section segmenting of TEM images (by hand) so that they can reconstruct a 3-D image. Please let me know if you'd like more info. Alice.
Alice Dohnalkova Environmental Microbiology Battelle, Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
Hello to all in the microscopy community,
I have previously posted a request for information on 3D reconstruction from serial sections.....but I only recieved replies from vendors and people wanting a summary of the replies.
{ { {I was wondering what computer programs people are using for 3-D reconstruction of TEM serial sections. I am interested in peoples opinions of the software (i.e. ease of use, learning curve, cross-platform functionality, cost, etc.). Feel free to email me off list if you choose. Also vendors feel free to send me any info you might have on the products you sell.} } }
I would really like to hear from the users of these programs! I would be glad to submit a summary to all interested parties (yes I still have your addresses).
Thanks to all,
-EK
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
see our web page at http://www.mayo.edu/mcniven_lab/
Microscopy Suite, room B650J Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana IL 61801 217 265-5071 (office); 217 244-6219 (fax) sjrobin-at-uiuc.edu
I emailed that we have an Amray 1000A SEM and a PGT EDS with Sun workstation that we do not need. They are available to anyone who would like to have them and will take care of taking them away. They are listed on the MSA web site under surplus equipment.
I have a Nikon Optiphot 88 microscope which doesn't give a good image. I have had Nikon representative in and got little help to improve the situation. So I am looking for an independent expert to check the microscope out and repair it. In addition, I'd like to add some more features such as dark field and possibly add couple of new lenses.
If someone know companies or individuals who provide that kind of service, please let me know.
Thanks in advance.
Hong Wang Principle Materials Scientist Guardian Science and Technology Center 14511 Romine Road Carleton, MI 48117
We have an Oxford CT 1500HF Cro Transfer System on our SEM and have blown the nude ion gauge. We have no information on the part , description or Part no.
If any one has one of these units, or can provide information from their manual. I would be very grateful to hear from them.
Many thanks
--
Angus Netting CEMMSA (Centre for Electron Microscopy & MicroStructure Analysis) Adelaide University Ph: 61 8 8303 5855 Fax: 61 8 8303 4356 http://www.adelaide.edu.au/CEMMSA
We use HMDS to "dry" SEM specimens. My bottle advises "Moisture sensitive. Stored under argon" Up to now I have used bottles stored in the fridge for up to a year.
1. Does anyone take any precautions to keep their HMDS dry?
2. Furthermore how long would you regard an opened bottle as useful?
3. We use molecular sieves to keep ethanol and acetone dry - should we put it in HMDS?
Dave
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
} } We need to purchase a new water recirculator for our electron microscope } (Philips CM-10). Is there a consensus of opinion on what the best } recirculator is, reliability being the highest consideration? Thanks. } } Vachik Hacopian ****************
I have a 21 year-old Haskris for my JEOL. It has needed very little attention:, mostly water changes when the engineers do my PMs. I had to replace the compressor 10 years ago, and just had a leak in the refrigerant tubing repaired.
(No financial interest in Haskris or JEOL)
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I have watched this thread and have refrained from posting since I did not want to "advertise" our products, but I believe that there is some erroneous information now.
Alice Dohnalkova said: "If the purpose is to produce 3-D modeling geometry (like what we want to do), then there is nothing available to his knowledge". Although I am not sure, what "3-D modeling geometry" entails, there is software that takes images, allows you to define certain features in each image of a stack (by hand or semiautomatically), and take all that information from the different slices and create a 3-D model of whatever was sliced. Of course you can then do measurements on the reconstruction. There are several of those packages out there (our 3D module is one of them). The other type of software is "voxel" based. In essence a stack of images is treated as a "cube" of information, which can then be sliced and lookt at in different directions.
The first option is normally used to recreate a 3-D representation of the object, which can then be further analyzed with regards to it's position relative to other objects or for certain 3-D parameters (volume, surface, 3D angles, etc.).
The second type (voxel-based) is normally used to create a 3-D information cube of the entire stack, then take slices in different directions and analyze these (artificial) images.
Both usually require that you know the separation between slices (from your microtome settings), and that you can apply a number of operations before creating the 3D shape to account for shift, rotation and distortion. It is sometimes necessary to embed a "standard" in the sample to be able to do that.
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Dohnalkova, Alice [mailto:Alice.Dohnalkova-at-pnl.gov] Sent: Tuesday, September 25, 2001 2:40 PM To: 'Krueger, Eugene W.'; 'Microscopy-at-sparc5.microscopy.com'
Eugene,
our group is involved in developing a program for 3-d reconstruction of TEM serial sections for modeling processes in prokaryotic cells. I asked the person responsible for the images stacking part (not a microscopist), and will try to translate his answer: It depends on what someone would want to do with the reconstruction. If you wanted to just look at the image of the reconstruction, then there is some software that you can kind of do this (with a lot of fiddling of the stack of data to get it into the program). The two packages are IBM's OpenDX and the VTK toolkit. If the purpose is to produce 3-D modeling geometry (like what we want to do), then there is nothing available to his knowledge. He says this is partly because there is no 3-D modeling code available that could consume such data. I do know of two groups that are doing section-by-section segmenting of TEM images (by hand) so that they can reconstruct a 3-D image. Please let me know if you'd like more info. Alice.
Alice Dohnalkova Environmental Microbiology Battelle, Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
Hello to all in the microscopy community,
I have previously posted a request for information on 3D reconstruction from serial sections.....but I only recieved replies from vendors and people wanting a summary of the replies.
{ { {I was wondering what computer programs people are using for 3-D reconstruction of TEM serial sections. I am interested in peoples opinions of the software (i.e. ease of use, learning curve, cross-platform functionality, cost, etc.). Feel free to email me off list if you choose. Also vendors feel free to send me any info you might have on the products you sell.} } }
I would really like to hear from the users of these programs! I would be glad to submit a summary to all interested parties (yes I still have your addresses).
Thanks to all,
-EK
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
see our web page at http://www.mayo.edu/mcniven_lab/
Several people have gotten back to me re: the website for the newsletter. The current (Sept. 2001) newsletter should be posted very soon on NESM's website--we had some problems with the site! I apologize for any inconvenience.
The meeting will begin with registration at 5-5:30pm at JEOL USA, Peabody. The demonstrations will be first and the technical talks will begin between 7-7:30pm.
Once the newsletter is posted--specific details will be listed. Please contact Mary McCann, NESM Treasurer, by email: mccanns-at-tiac.net to pre-register for this meeting.
Peggy Sherwood Corresponding Secretary, NESM -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
To the person asking about water chillers: I have a Haskris water cooled water chiller that has run for 15 years with out failure. I think that's about as reliable as a machine can get. Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 JAH 1020 Locust Street Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cell Timothy.Schneider-at-Mail.TJU.edu
A researcher has asked if he can examine cell fractions from a gel to determine which organelles comprise each band in the gel. The bands can be sucked off and isolated in suspension. Are there any quick techniques to determine which band is mitochondria, nuclei, etc. I have a Hticahi S3500N tungsten filament SEM and a cryo stage. How about a negative stain and a quick look in the TEM?
I too have high regard for Haskris. We had one that was over 35 years old and still kicking when we moved to a new lab. We now have a younger one (maybe 25 yrs old) and a new 2-tank one (2 yrs). They're very reliable.
..no financial interest in this company, just a satisfied customer.
Sara
On Wed, 26 Sep 2001, Leona Cohen-Gould wrote:
} Date: Wed, 26 Sep 2001 09:34:24 -0400 } From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} } To: Vachik Hacopian {vhacopian-at-wellesley.edu} , } microscopy-at-sparc5.microscopy.com } Subject: Re: water recirculators for electron microscopes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } We need to purchase a new water recirculator for our electron microscope } } (Philips CM-10). Is there a consensus of opinion on what the best } } recirculator is, reliability being the highest consideration? Thanks. } } } } Vachik Hacopian } **************** } } I have a 21 year-old Haskris for my JEOL. It has needed very little } attention:, mostly water changes when the engineers do my PMs. I had } to replace the compressor 10 years ago, and just had a leak in the } refrigerant tubing repaired. } } (No financial interest in Haskris or JEOL) } } Lee } } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
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Our system is unique - for the first time, we have combined the power of the Internet with the ease of the telephone to enable immediate access to a psychologist.
How does it work? Using our revolutionary new technology, you can reach our doctors by clicking on this link: http://dev.pnonline.com/EMReturn.asp?EGUID=10D7096B-CC89-4DAA-A069-F6E781DDAC28&ELinkID=191. Simply enter your callback phone number and credit card info and you will receive an immediate callback from one of our doctors! Or, just pick up the phone and call 1-877 DR TALKS (1-877 378-2557)!
Get expert advice on such problems as :
parenting
relationships
stress
sexuality
more.....
Our service is convenient - we are available 24/7/365.
*Promotional Special: A cruise for two to the Caribbean will be awarded to one registered client. The winner will be contacted via email. Check our Website for details.
If you would no longer like to be a part of our mailing list, please accept our apologies in advance for this email. To be removed, just click this link: http://dev.pnonline.com/unsubscribe.asp?EGUID=10D7096B-CC89-4DAA-A069-F6E781DDAC28&ELinkID=192.
Anybody wanna sell their Ultracut E? Please respond to Christina Stamper. "Bennett-stamper, Christina (BENNETCN)" {BENNETCN-at-UCMAIL.UC.EDU} } You can see I am at a new e mail address as I have just started at the } Univ of } Cincinnati. I am still looking for an ultracut E. I just wanted } to let you know } just in case you hear anything...thank you so much. } Christina Stamper University of Cincinnati EMF operations manager
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
We are looking for a source of larger sizes of EM grid material. We are presently using 750 and 1000 mesh grids in 1" squares from SPI. We would like to get it in 3"x4" size, even if we have to cut it down from a larger size. SPI says they can supply us with 6x6" but they don't normally stock that and it takes some time to get it. We would be willing to cut it down. Does anyone know of other sources of this material? We have tried most of the standard EM materials suppliers, and they have referred us back to SPI. Any ideas?
Thanks in advance,
Bert Gough, Ph.D. Vice President, Systems R&D PH: (412) 826-3600 X-2644 Cellomics, Inc. FX: (412) 826-3850 635 William Pitt Way EM: agough-at-cellomics.com {mailto:agough-at-cellomics.com} Pittsburgh, PA 15238 Visit CellSpace(TM) Knowledge Miner: Automated Intelligence for the New Biology http://cellspace.cellomics.com {http://cellspace.cellomics.com}
I did a search on the internet for "electroformed mesh" because I could remember the name of the company that I bought my larger sized grids from. The company is Buckbee Mears. I think that they are also a source of EMP protection for mesh on some of our laminate products for transparencies military aircraft. I have copied a paragraph from one page that has some of the information that you might need to contact them. I once had a catalog from their company, but that was long ago.
BMC Industries, Inc. is a multinational manufacturer and distributor of a variety of products through two segments: Buckbee-Mears and Optical Products. Buckbee-Mears is comprised of Mask Operations and Buckbee-Mears St. Paul (BMSP). Mask Operations produces aperture masks, a critical component of color television and computer monitor picture tubes. BMSP is the leading producer of precision photo-etched metal and electroformed parts. Optical Products designs, manufactures and distributes polycarbonate, glass and hard-resin plastic ophthalmic lenses.
Here is their web address: http://www.bmcind.com/
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Bert Gough [mailto:agough-at-cellomics.com] Sent: Thursday, September 27, 2001 8:42 AM To: microscopy-at-sparc5.microscopy.com Cc: Anika Joseph; Alex Friedman
We are looking for a source of larger sizes of EM grid material. We are presently using 750 and 1000 mesh grids in 1" squares from SPI. We would like to get it in 3"x4" size, even if we have to cut it down from a larger size. SPI says they can supply us with 6x6" but they don't normally stock that and it takes some time to get it. We would be willing to cut it down. Does anyone know of other sources of this material? We have tried most of the standard EM materials suppliers, and they have referred us back to SPI. Any ideas?
Thanks in advance,
Bert Gough, Ph.D. Vice President, Systems R&D PH: (412) 826-3600 X-2644 Cellomics, Inc. FX: (412) 826-3850 635 William Pitt Way EM: agough-at-cellomics.com {mailto:agough-at-cellomics.com} Pittsburgh, PA 15238 Visit CellSpace(TM) Knowledge Miner: Automated Intelligence for the New Biology http://cellspace.cellomics.com {http://cellspace.cellomics.com}
The EM supply houses carry special (clean & without grit) styrofoam sticks for cleaning diamond knives; these are fairly inexpensive. I'm sure these would be a good source for scratch-free styrofoam with which to clean lenses.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
A few more notes about the CSMMS meeting on 10/19. If you plan on attending, please contact me (see below) by October 12. There is no registration fee for the meeting, but we need to know how many attendees because lunch will be catered. The lunch is only $5. And if needed, I can send you a list of hotels and a map for directions.
Caroline Miller will be bring the MSA Traveling Poster Exhibit, Caroline Schooley will be providing information on Project MICRO and there will be a live TelePrescence Microscopy connection. There will also be an area for our Corporate Sponsors and other vendors to discuss their products and services.
Hope to see you in Columbia.
Lou ____________________
Central States Microscopy and Microanalysis Society Fall Meeting Friday, October 19, 2001 Adams Conference Center Veterinary Medicine Building University of Missouri Columbia, Mo 65211
8 - 8:45 Registration (Free)
8:50 Welcome
9:00 Tobias Baskin, University of Missouri-Columbia, "Ultrastructure of the Plant Cell Wall Analyzed with Field-Emission Scanning Electron Microscopy"
9:15 Nadia Navarrete-Tindall, University of Missouri-Columbia, "Morphology of the Endangered Legume Amorpha nitens Boynton and its Rhizobia Symbiont"
9:30 Heide Schatten, University of Missouri-Columbia, "Cytoskeletal Dynamics in Apicomplexan Parasites"
9:45 Joe Simmons, University of Missouri-Columbia, "Idiopathic Interstitial Pneumonia in Laboratory Rats"
10:00 Marty Katz, University of Missouri-Columbia, "Stem Cell Transplantation Therapy for Inherited Neurodegenerative Disorders"
10:15 Tom Phillips, University of Missouri-Columbia, "Fixation of Biological Tissues - Science or Art?"
10:30 - 10:45 BREAK
10:45 Robert Hall, Associate Vice Provost of Research, University of Missouri-Columbia, "The University of Missouri: Taking the Lead in National Research Funding"
11:00 Keynote speaker: Kenneth Moore, University of Iowa, MSA Tour Speaker, "Applications of Microscopy Techniques to the Study of Genetic Therapy Research"
12:00 - 1:00 Lunch
1:00 - 1:30 CSMMS Business Meeting
1:30 John Bozzola, SIU-Carbondale, "Multidisciplinary Approach to Teaching Electron Microscopy"
1:45 Howard Wilson, University of Missouri-Columbia, "Creating a Scientific Poster Presentation using Microsoft Powerpoint"
2:00 Scott Walck, PPG, "Low Angle Cleavage - A New Specimen Preparation Method for TEM"
2:15 Louis Ross, University of Missouri-Columbia, "Exciting Times in Scanning Electron Microscopy and X-ray Microanalysis"
2:30 Vladimir Dusevich, University of Missouri-KC, "Practical ESEM"
2:45 Jeff Speakman, University of Missouri-Columbia, "Laser Ablation ICP-MS as a Tool for Characterization of Archaeological Materials"
3:00 - 3:15 BREAK
3:15 Cammy Bright, University of Missouri-Columbia, "A Laser Ablation ICP-MS and SEM Study of Rare Earth and Trace Elements in Conodonts"
3:30 Mike Amspoker, Westminster College, "A Light and Scanning Electron Microscopy Study of the Freshwater Diatom Desmogonium rabenhorstianum Grunow"
3:45 Heather Ramsay, University of Missouri-Columbia, "New Applications of SEM in Osteological Morphology Research"
4:00 Dickerson Moreno, University of Missouri, "Development of N-Type Diamond Semiconductor Through Field Enhanced Diffusion by Optical Activation"
4:15 Alejandro Suarez, University of Missouri, "A New Method to Fabricate Polycrystalline Diamond as a P-Type Semiconductor"
4:30 Closing Remarks _______________________
Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.bioterc.missouri.edu/emc
I just came across a web site that may help this discussion. I know that I can't address the needs of others, but the Wadsworth Center appears to be serious about the methods and the approach. URL: http://www.wadsworth.org/rvbc/index.html
Also the HVEM at Boulder has a 3-D Subdivision with software available: URL: http://bio3d.colorado.edu/index.html
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
We have recently put a digital camera on our TEM. We have worked out most of the problems related to obtaining accurate and precise measurements from the digital images on the monitor. Now I must deal with printer paper.
I believe we have 2 different printing needs. First, we will be printing hundreds of TEM images per year that do not have to be high quality but will be filed away for 10-20 years. Second, we will hopefully need to print several publication quality prints per year for submission with manuscripts. We have an EPSON Sylus 980 ink jet printer and a long list of possible printer papers. Before I buy many different papers to test, I would appreciate opinions regarding which papers will fill our 2 needs.
Thanks for your help,
John
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
You may have to find better ink that what is normally supplied if you want the images to last years, or get an (relatively) inexpensive laser printer like the NEC. Even 600dpi yields decent images from our Philips XL 20. I believe the newer model NEC (870?) does much better .
Ron ----- Original Message ----- } From: "John Basgen" {basgen-at-maroon.tc.umn.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, September 28, 2001 9:34 AM
The Biophysical Discussions ........................................................................... .......... Frontiers in Structural Cell Biology: Determining the structures of large subcellular machines ........................................................................... .......... Meeting Dates: April 20-22, 2002.
Meeting Site: Asilomar, California. This is a spectacular site. You can see it at http://www.asilomarcenter.com/. ........................................................................... .......... The biophysical discussions are set up so that the discussion time is greater than the times allotted for presentations, which will be available on the web prior to the meeting. The format is unusual, but highly successful in the past.
To apply send a letter of intent to discussions-at-biophysics.org by November 15, 2001. Space is limited. ........................................................................... .......... Organizing committee: Axel Brunger (Stanford University), David DeRosier (Brandeis University), Steve Harrison (Harvard University), and Eva Nogales (University of California at Berkeley).
The Program (Saturday, April 20 at 9 am to Monday April 22 at 12 noon)
Session I. The state of structural biology of large structures. Moderator Helen Saibil Richard Henderson The power of electron cryomicroscopy. Joachim Frank The Ribosome -- a molecular machine in motion Jamie Cate Biochemical basis for x-ray crystallography of the ribosome
Session II. Extending x-ray crystallography to ever larger structures Moderator Keith Hodgson Andy Thompson Can we routinely collect useful data from micro-crystals? Janos Hajdu Future X-ray sources (tentative) Randy Reed The phase problem: does size matter?
Session III. New ways to obtain large complexes for structural studies Moderator Axel Brunger Don Wiley Stabilizing multi-component biological complexes for structural studies by protein engineering, expression, and refolding - AND - avoiding artifacts TBA Expression and Co-expression of components (tentative title)
Session IV. What does the future hold for electron cryomicroscopy? Moderator Bob Glaeser Niko Grigorieff Single particles always fit the mold Ken Downing The hybrid approach to electron crystallography Wolfgang Baumeister Electron Tomography: Towards visualizing macromolecular assemblies inside cells Ed Egelman Polymorphism, can we detect it? Can we use it? Can we control it? Examples from actin and nucleoprotein complexes.
Session V. Can hybrid methods provide credible atomic models? Moderator Eva Nogales Niels Volkmann Atomic model of the cell: docking in a tomographic environment Willy Wriggers Reconciling Shape with Structure: Morphometric Strategies for Multi-Resolution Flexing
Hi John, The goal of linear, archival, high-quality gray scale images with a large dynamic range can be difficult to achieve. One option is a Codonix printer - they produce pretty stunning photo-quality prints, but they are pretty expensive. Another option might be to go with a quad-black print system. There are several available for Epson ink jet printers, and I have had good success with one of these (sold by MIS Associates) set ups on an Epson 1200. You might want to check out InkJet Mall (http://www.inkjetmall.com/store/index.html) , and see what they have to offer. I am not affiliated with them, but ran across them as a vendor. They work with the art industry, and have developed a proprietary photoshop pluggin, printer driver, and inkset that is supposed to give outstanding gray scale images. I just ordered a system, and it will be arriving shortly. Its called piezography BW. As far as papers go, you might want to check out LumiJet Preservation Series Gallery Gloss. I've tested a number of different papers with pigmented inks, and that's the one that I liked the best. Some vendors that I have used in the past are: InkjetArt.com at: http://www.tssphoto.com/sp/dg/luminos/index.html, and MIS Associates at: http://www.inksupply.com/
Good luck.
-Brad
---------- From: John Basgen Sent: Friday, September 28, 2001 06:34 To: Microscopy-at-sparc5.microscopy.com Subject: TEM Ink Jet Printer Paper
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Microscopists,
We have recently put a digital camera on our TEM. We have worked out most of the problems related to obtaining accurate and precise measurements from the digital images on the monitor. Now I must deal with printer paper.
I believe we have 2 different printing needs. First, we will be printing hundreds of TEM images per year that do not have to be high quality but will be filed away for 10-20 years. Second, we will hopefully need to print several publication quality prints per year for submission with manuscripts. We have an EPSON Sylus 980 ink jet printer and a long list of possible printer papers. Before I buy many different papers to test, I would appreciate opinions regarding which papers will fill our 2 needs.
Thanks for your help,
John
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
Hello! I would like to settle an on-going debate about pre-perfusion with saline. Does anyone pre-perfuse with saline before perfusion with fixative for CNS tissue? Thank you for your help. Meghan Towner University of Idaho
We are looking for a used ultramicrotome for our materials science lab. If anyone has one they are looking to see, or if anyone knows where to find one, could you please contact me.
Thanks,
Praveena Bhaskara
Graduate student
Department of chemical engineering
University of Massachusetts,Lowell
978-934-3411
Praveena Bhaskara
Get your FREE download of MSN Explorer at {http://go.msn.com/bql/hmtag_itl_EN.asp} http://explorer.msn.com
For my TEM digital system, I have a HP 960 inkjet printer ($250.00) with 2600 x 1200 dpi quality. The paper I am currently using is HP Premium Paper at ~$50.00 for 50 sheets. I have tried to get a better deal by purchasing in bulk, but I was told by our supplier (Corporate Express) Hp does not sell in bulk and only sell the pre-packaged 50 sheet units.
I am still in the process of evaluating other products/manufacturers of photo quality paper.
Ken
-- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Director, Imaging Center Legacy Holladay Park Medical Center 1225 NE 2nd Avenue Portland, OR 97232
On Fri, 28 Sep 2001, John Basgen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Microscopists, } } We have recently put a digital camera on our TEM. We have worked out most of } the problems related to obtaining accurate and precise measurements from the } digital images on the monitor. Now I must deal with printer paper. } } I believe we have 2 different printing needs. First, we will be printing } hundreds of TEM images per year that do not have to be high quality but will be } filed away for 10-20 years. Second, we will hopefully need to print several } publication quality prints per year for submission with manuscripts. We have an } EPSON Sylus 980 ink jet printer and a long list of possible printer papers. } Before I buy many different papers to test, I would appreciate opinions } regarding which papers will fill our 2 needs. } } Thanks for your help, } } John } } John M. Basgen } Department of Pediatrics } University of Minnesota } Mayo Mail Code 491 } 420 Delaware Street SE } Minneapolis, MN 55455 } USA } Phone: 612-625-7979 } FAX: 612-626-2791 } E-mail: basgen-at-umn.edu } }
For high-definition ink-jet paper try Jet Print Imaging from International Paper. It is better the HP Premium and cheaper. It comes in reams (only) of 500 at $175, which $0.35/sheet. Cadmet is one of the distributors.
JQuinn
PS: They will also send you a sample pack.
} From Microscopy-request-at-sparc5.microscopy.com Sat Sep 29 09:32:49 2001 } Date: Sat, 29 Sep 2001 02:17:14 -0700 (PDT) } From: Ken Tiekotter {tiekotte-at-up.edu} } To: John Basgen {basgen-at-maroon.tc.umn.edu} } cc: {Microscopy-at-sparc5.microscopy.com} } Subject: Re: TEM Ink Jet Printer Paper } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear John, } } For my TEM digital system, I have a HP 960 inkjet printer ($250.00) with } 2600 x 1200 dpi quality. The paper I am currently using is HP Premium } Paper at ~$50.00 for 50 sheets. I have tried to get a better deal by } purchasing in bulk, but I was told by our supplier (Corporate Express) Hp } does not sell in bulk and only sell the pre-packaged 50 sheet units. } } I am still in the process of evaluating other products/manufacturers of } photo quality paper. } } Ken } } } -- } Ken Tiekotter, Adjunct Professor } The University of Portland } Department of Biology } 5000 N. Willamette Blvd. } Portland, OR 97203 } } Director, Imaging Center } Legacy Holladay Park Medical Center } 1225 NE 2nd Avenue } Portland, OR 97232 } } } On Fri, 28 Sep 2001, John Basgen wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Microscopists, } } } } We have recently put a digital camera on our TEM. We have worked out most of } } the problems related to obtaining accurate and precise measurements from the } } digital images on the monitor. Now I must deal with printer paper. } } } } I believe we have 2 different printing needs. First, we will be printing } } hundreds of TEM images per year that do not have to be high quality but will be } } filed away for 10-20 years. Second, we will hopefully need to print several } } publication quality prints per year for submission with manuscripts. We have an } } EPSON Sylus 980 ink jet printer and a long list of possible printer papers. } } Before I buy many different papers to test, I would appreciate opinions } } regarding which papers will fill our 2 needs. } } } } Thanks for your help, } } } } John } } } } John M. Basgen } } Department of Pediatrics } } University of Minnesota } } Mayo Mail Code 491 } } 420 Delaware Street SE } } Minneapolis, MN 55455 } } USA } } Phone: 612-625-7979 } } FAX: 612-626-2791 } } E-mail: basgen-at-umn.edu } } } } } } } }
If you're referring to part number C6979A, HP premium glossy photo paper, it is available for less than half that at many retailers.
Regards, Dave Harrison
On 29 Sep 2001, at 2:17, Ken Tiekotter wrote:
} } For my TEM digital system, I have a HP 960 inkjet printer ($250.00) with } 2600 x 1200 dpi quality. The paper I am currently using is HP Premium } Paper at ~$50.00 for 50 sheets. I have tried to get a better deal by } purchasing in bulk, but I was told by our supplier (Corporate Express) Hp } does not sell in bulk and only sell the pre-packaged 50 sheet units. }
Hi All, I'm trying to identify Helicobacter pylori in the dental plaque using IHC (immunoenzymatic/immunohistochemicals) staining techniques with primary antibody from rabbit. For the observations I'm using SEM thanks to the reaction between oxidated DAB and Osmium that forms an electondense compound.Unfortunately the adherence of bacterium to glass seems to be an insurmountable obstacle. Does somebody know any quick and strong techniques to "stick" on glass cover slip the dental plaque? thancks in advance T. Train
Hi All, I'm trying to identify Helicobacter pylori in the dental plaque using IHC (immunoenzymatic/immunohistochemicals) staining techniques with primary antibody from rabbit. For the observations I'm using SEM thanks to the reaction between oxidated DAB and Osmium that forms an electondense compound.Unfortunately the adherence of bacterium to glass seems to be an insurmountable obstacle. Does somebody know any quick and strong techniques to "stick" on glass cover slip the dental plaque? thancks in advance T. Train
Hi All, I'm trying to identify Helicobacter pylori in the dental plaque using IHC (immunoenzymatic/immunohistochemicals) staining techniques with primary antibody from rabbit. For the observations I'm using SEM thanks to the reaction between oxidated DAB and Osmium that forms an electondense compound.Unfortunately the adherence of bacterium to glass seems to be an insurmountable obstacle. Does somebody know any quick and strong techniques to "stick" on glass cover slip the dental plaque? thancks in advance T. Train
The Boston University Physics Department is upgrading its Materials Research Facility and looking for a buyer for its existing STEM. Please contact: Prof. Bennett Goldberg Boston University Physics Department 617-353-5789 goldberg-at-bu.edu
------------------------------------- Philips CM12 (120 keV) "Twin Lens" Electron Microscope with Software Upgrade, S/N D733 purchased 1988 STEM Unit Model 5227 (for CM10/CM12), S/N D733.1 Specimen Relocation System Model 5235, S/N D733.2 Backscatter detector (for selecting high Z) Secondary electron detector (for 5-nm-imaging) Gatan film camera (for conventional "photography") Various Gatan sample holders Haskris Cooling Water Heat Exchanger EDAX Energy Dispersive Analyzer, Model PV9800, S/N HX-626/01
The microscope was used for conventional electron microscopy, 5-nm-resolution Scanning Electron Microscopy, and 10-nm resolution electron-beam lithography. The lithography limitations were set by the maximium chip size (approx. 5 mm by 8 mm), the typical field of view while writing (80 micrometers), and the mechanical relocation alignment accuracy (0.25 micrometers).
The system does not have a beam blanker, but has been equipped with a carefully shielded "x-y" input. Our homemade pattern definition system is obsolete and not included.
The microscope was under service contract, fully operational, and checked out by the Philips/FEI preventative maintenance service several times each year until it was decommisioned last year.
--- Bennett B Goldberg Professor of Physics Boston University 590 Commonwealth Ave Boston, MA 02215 (617) 353-5789/ 1712 lab/ 9917 fax goldberg-at-bu.edu http://ultra.bu.edu
from my experience with mouse (CNS; spinal chord, muscle, testis and other tissue) I would strongly recommend a short (!) pre- perfuse (as you name it) to flush out all blood cells, which otherwise regularly clog and occlude the fine capillaries. However, though simple saline flush can be sufficient, I routinely use a mixture of Heparin (anti-coagulant), Procain (keeps the blood vessels "open"), PVP (polyvinylpyrrolidon, MW-class 30 K; for osmotic pressure) in aqua dest. and pH-adjustment. Method published originally in a wonderful perfusion-technique paper by Forssmann, WG et al (1977) in Anat. Rec. 188: p 307- 314. Just follow their technical procedure exactly and you`ll get excellent fixation.
good luck!
peter
************************************************** please reply to: peter.heimann-at-biologie.uni-bielefeld.de ("E-MAIL")
Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : xx49(0)521-106-5654 WEB-Site: http://www.uni-bielefeld.de/biologie/Entwicklungsbiologie WEB-Site: http://www.uni-bielefeld.de/SFB549 ******************************************************
Sorry for being rather late with my reaction, but – as far as I have seen – nobody has given a hint relating the home of “3-Dimensional Electron Microscopy” (http://3dem.sdsc.edu/). This page collects information about the laboratories and services available to the 3D-EM community. A lot of available reconstruction packages are listed at this page. Additionally there exists also a listserver for specific questions/problems.
I hope this helps.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Herbststrasse 7 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: http://www.tvips.com/ Email: ingo.daberkow-at-tvips.com +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
"Krueger, Eugene W." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello to all in the microscopy community, } } I have previously posted a request for information on 3D reconstruction from } serial sections.....but I only recieved replies from vendors and people } wanting a summary of the replies. } } { { {I was wondering what computer programs people are using for 3-D } reconstruction of TEM serial sections. I am interested in peoples opinions } of the software (i.e. ease of use, learning curve, cross-platform } functionality, cost, etc.). Feel free to email me off list if you choose. } Also vendors feel free to send me any info you might have on the products } you sell.} } } } } I would really like to hear from the users of these programs! I would be } glad to submit a summary to all interested parties (yes I still have your } addresses). } } Thanks to all, } } -EK } } Eugene W. A. Krueger } Sr. Research Technologist in Cell Biology } Center for Basic Research in Digestive Diseases } Mayo Clinic and Foundation } (507)284-0580 (lab) } (507)284-0762 (fax) } krueger.eugene-at-mayo.edu } } see our web page at http://www.mayo.edu/mcniven_lab/
The only two brands I've had personal experience with are Haskris and Neslab chillers. Both seem pretty good - usually the first problem you run into (hopefully only after several years' service) is the pump failing. In these two brands the pumps are often identical, made by a third company. I think I prefer the Haskris machines, though - their reservoirs tend to be readily openable for inspection - most seem to have square tanks with a lid that you can easily lift off to see what's going on inside. The Neslab chiller we have now has a cylindrical, sealed tank, so it can be a bit more difficult to troubleshoot. Hope this helps.
Frank Thomas GSC Atlantic Bedford Institute of Oceanography Dartmouth, Nova Scotia ----- Original Message ----- } From: "Vachik Hacopian" {vhacopian-at-wellesley.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, September 25, 2001 3:24 PM
Dear Fellow Microscopists;
I have a question relative to the detection of Tl [Thallium] in the presence of a Au plated surface. The application of this plated region is the metallization system of a microcircuit. SIMS analysis was performed on these Au plated surfaces and found the presence of thallium in the several atm. %. However, when I attempt to detect it using EDX, I cannot. I have looked at the energy spectra and found that they separate out well in the higher energies yet I see absolutely no indication of thallium. The thickness of the sample is ~ 1.0 uM and about 100 um X 100 uM in area. Plenty enough for this analysis.
Does anyone know of any unique situation with thallium in gold? Either the SIMS analysis is in error or I'm overlooking something.
Peter Tomic Analytical Services Anadigics, Inc. Warren, New Jersey
Check out ASTM standards. ...Available for both Quant EDS and Magnification for SEMs.
Woody
} -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Does anyone know of any documented EDS calibration (energy display, } quantification, etc.) procedures that have been certified in any way? } i.e. NIST, ISO-9000 or any other. } } thanks, } } Mark Wall } LLNL } 925 423 7162 }
Opportunities for Postdoctoral Fellows in Electron Cryomicroscopy and Structural Biology Prof. David DeRosier (derosier-at-brandeis.edu). Rosenstiel Basic Medical Sciences Research Center Brandeis University
I have a postdoctoral position to study the structure of large macromolecular machines. The position is fully funded for at least two years, and we will pay competitive salaries. Write me if you or someone you know is interested. Here is a brief description of the available projects:
The cytoskeleton: The actin cytoskeleton is a set of cellular machines responsible for many of the dynamic capabilities of eukaryotic cells. Actin bundles are a key elements used to shape cells, generate filopodia, and order the cell cytoplasm. The project is aimed at determining the structure of the actin bundle in the brush border cells, as a model system. These bundles are the best characterized and most tractable bundles. The approach is two pronged beginning with in vitro studies of the components (actin, fimbrin and villin) and leading to in vivo studies of intact bundles. The tool we use is electron cryomicroscopy and image analysis.
The bacterial flagellum: The flagellum is a several machines in one. It is composed of over a dozen different proteins, with one set making up the rotary motor, another the drive train, another a bushing and seal, and another the flagellar export apparatus. We determining the structures of these components using electron cryomicroscopy and image analysis, and we are correlating structural features with the primary sequences of the component proteins.
The bacterial signaling complex: The bacterial cell senses the environment and transmits a signal to the flagellum. We are using electron cryomicroscopy and image analysis to determine the structure of a 1.4 megadalton complex of the cytoplasmic domain of a chemoreceptor, a kinase, and an adaptor molecule. The complex is highly active enzymatically, but has an unusual stoichiometry, which suggests signal integration from several receptors into a single kinase.
Professor David J. DeRosier Abraham S. and Gertrude Burg Chair of Life Sciences W.M. Keck Institute for Cellular Visualization Rosenstiel Basic Medical Sciences Research Center MS029 Brandeis University 415 South Street Waltham, MA 02454-9110 Telephone: 781-736-2494 FAX: 781-736-2405 email: derosier-at-brandeis.edu
I need to find someone in the Washington, DC area (or close by) who repairs RMC ultramicrotomes. We have an MT-7 that has a very fine chatter visible only under the beam.
If you know of anyone/company that does this type or work, please let me know.
I don't mind chatter, just not on my sections.
Paula :-)
p.s. hey sectioner, sectioner.....Breathe!
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We are looking for a specimen rotating holder for a JEOL 100CX part# IEM100/200CX-SRH. Also for the JEOL 100CX we are in need of a dispensing magazine, receiving magazine and/or cassette film holders. If you are looking for a home for any of these items please contact me.
Thank you,
Ruth Yamawaki Department of Comparative Medicine Stanford University (650) 723-3457 FAX (650) 498-6259
Hello All I need to purchase a new 3mm / 5mm Diamond knife for ultrathin sections of calcified tissue and polyurethane samples in Spurs resin. The prices from most of the suppliers are horrendous, as you well know. Does anyone out there have any of these lying around which are not used? As a last resort we'll consider a good, used one.Any brand name would do. And where can I get my "old" one re-sharpened at a reasonable price? Thanks and regards Nazlia
I am looking for a high end deconvolution program that would also include image acquisition capabilities. I colleague would like to do both with a single software package.
Any suggestions would be appreciated. Thank you.
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8 Canada
Paula: For many years we have relied on two firms for ultramicrotome maintenance and repair work. Both firms we found could be counted on for prompt and excellent work. For our MT-series ultramicrotomes we have engaged the services of Bill McGee of Microtome Service Co. (315)-451-1404. For our Reichert and LKB instruments we relied on Helmut Ptzig and Mario Saccoccio of Microscopical Optical Consulting Inc. (914) 268-6450 or e-mail MOCLeica-at-aol.com.
Its possible that the chatter is not being generated from the ultramicrotome, but from some outside source of vibration. If so, you may want to install an anti-vibration platform. Regards, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences State University of New York-Binghamton Binghamton, NY 13902-6000
I have inherited a 1980s JEOL X-Ray Counting Unit Type XCU, which I want to press into service as a ratemeter for wds alignment purposes.
Is anyone out there sufficiently familiar with this model and sufficiently generous with their time to reply to me off-list so that I can be enlightened?
thanks
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Diamond knife manufacturers are mostly located in Switzerland and USA and they will tell you that prices have remained constant or decreased over the past 20 years. But you are right too, countries whose currencies have not kept pace with the US$, the Swiss Franc and the Euro find diamond knives and many other imported items very expensive indeed. The Australian dollar 2 years ago was equal to US 65 cents. Now it is 48.5 cents; a decline of 25% at a time of lower inflation here than in the USA. When our business was started 22 years ago we received US$1.20 for one A$. This topic is peripheral to the listserver, but the exchange rate woes do affect numerous microscopists. There is nothing we can do, but to encourage microscopists in the high exchange rate countries to come and visit our country. Australia is cheap for you to travel in, as our $ buys as much of most Australian made goods as the US$ buys in the USA. We have an excellent conference planned in February 2002 in Adelaide. Spend your money here and support our A$ ! Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, October 02, 2001 8:42 PM, Nazlia [SMTP:ctssamodien-at-samiot.uct.ac.za] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello All } I need to purchase a new 3mm / 5mm Diamond knife for ultrathin } sections of calcified tissue and polyurethane samples in Spurs resin. } The prices from most of the suppliers are horrendous, as you well } know. Does anyone out there have any of these lying around which are } not used? As a last resort we'll consider a good, used one.Any brand } name would do. And where can I get my "old" one re-sharpened at a } reasonable price? } Thanks and regards } Nazlia
I can't find compagnies which have spin-polarization detectors (magnetic microstructures in SEM) in their sales program. Does anyone out there can help.
Ing. Rudy De Vos K.U.Leuven - Department MTM Kasteelpark Arenberg 44 B - 3001 Heverlee Belgium
} Good Morning, } Does anyone out there know why you add ammonium chloride to a pre } incubating step? Caroline Miller ****************** It quenches fluorescence from unreacted aldehydes.
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
you are trying to bind up all the free aldehyde groups left over from fixation with aldehydes so they don't react with your antibodies. glycine works too. tom
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear listers, Could someone please send me the confocal istserve URL for subscription to that list. Many thanks, Jeremy jb_sanderson-at-yahoo.com
____________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk or your free -at-yahoo.ie address at http://mail.yahoo.ie
Anyone know of a good repair facility in PA or close by for a Bausch & Lomb Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads.
Dave Stadden Armstrong World Industries, Inc. 717-396-5109 drstadden-at-armstrong.com
Does anyone give me informations about Oxford INCA 100 EDS file format? Any reference for an INCA {--} EMS spectra converter?
P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
As has been pointed out, ammonium chloride reduces aldehyde double bonds so they won't react with your antibodies. Another advantage of reducing double bonds is to cut down the amount of autofluorescence in your specimen (in other words, places that fluoresce nonspecifically, because of double bonds). Ammonium chloride is relatively gentle in this regard. If your specimen has strong autofluorescence, you may need to resort to a more drastic procedure involving sodium borohydride (which is toxic, so has to be handled carefully). A good article that deals with sodium borohydride and bond reduction is:
Eldred et al, 1983, Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry, J Histochem Cytochem 31:285-292.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Is there a procedure for determining the shading of a fluorescence microscope image? I assume that one needs to take an image of a uniform object, but are there standard objects? Will a volume object work, or must I use a surface object. Would a front-surface mirror work, assuming I could measure the leakage through the barrier filter? Everett Ramer
To my understanding (which is admittedly weak in this area) the preferred method for visualizing membrane structure is to use freeze-fracture, then carbon replica, then TEM. Does anyone have an opinion as to whether high resolution FE-SEM of metal-coated samples would be as effective in this regard as viewing a replica in the TEM.
Bob
Robert R. Wise, Ph.D. University of Wisconsin-Oshkosh
Currently on sabbatical at UW-Madison Botany Department 430 Lincoln Drive Madison, WI 53706 email: wise-at-uwosh.edu phone: 608-262-4288 fax: 608-262-7509
TekNet has fixed several of our binocs. Email Jon Petz and give him your model info, etc., to see if he can work on it. He's in your area.
jon-at-teknetinc.com
Sara
On Wed, 3 Oct 2001 David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com wrote:
} Date: Wed, 3 Oct 2001 12:01:31 -0400 } From: David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com } To: Microscopy-at-sparc5.microscopy.com } Subject: East coast repair for B&L scope? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Anyone know of a good repair facility in PA or close by for a Bausch & Lomb } Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads. } } Dave Stadden } Armstrong World Industries, Inc. } 717-396-5109 } drstadden-at-armstrong.com } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Try the Leica Inc. facility in Buffalo NY and ask for Ken Brown. I think he can help you. The phone number is (716)686-3000 or if needed the main Leica # 1-800-248-0123. Leica owns B&L and handles parts for these microscopes.
Dave Joswiak Dept. of Astronomy University of Washington Seatle, WA 98195 (206)543-7702
On Wed, 3 Oct 2001 David_R_Stadden-at-armstrong.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Anyone know of a good repair facility in PA or close by for a Bausch & Lomb } Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads. } } Dave Stadden } Armstrong World Industries, Inc. } 717-396-5109 } drstadden-at-armstrong.com } } } }
In INCA select the spectrum in any of the Navigator windows (Acquire, Confirm, Report etc.), right mouse click, then select 'Export'. You have the choice of image and file types inculding EMSA format.
Ron
On Wed, 3 Oct 2001 19:19:49 +0200 FABBRI {fabbri-at-cigssrv1.unimo.it} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anyone give me informations about Oxford INCA 100 EDS file format? } Any reference for an INCA {--} EMS spectra converter? } } P.L. Fabbri } } --------------------------------------------- } Dr. Pier Luigi Fabbri } C.I.G.S. - Centro Interdip. Grandi Strumenti } Università di Modena } via Campi 213/A - 41100 Modena, Italy } Tel +39-059-2055231 / +39-059-370551 } Fax +39-059-370551 } E-mail: fabbri-at-mail.cigs.unimo.it } --------------------------------------------- } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
We have been very happy with Paul Musci of M&R Optical. He is a former B&L serviceman. He can be reached at:
PO Box 515 Charlton City, Massachusetts 01508 508-248-4658
Helen Mayer UCAR Carbon Parma, OH
-----Original Message----- } From: "David_R_Stadden-at-armstrong.com"-at-sparc5.microscopy.com [mailto:"David_R_Stadden-at-armstrong.com"-at-sparc5.microscopy.com] Sent: Wednesday, October 03, 2001 12:02 PM To: Microscopy-at-sparc5.microscopy.com
Anyone know of a good repair facility in PA or close by for a Bausch & Lomb Stereozoom 7? I'm told the focusing rack is messed up. Appreciate any leads.
Dave Stadden Armstrong World Industries, Inc. 717-396-5109 drstadden-at-armstrong.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We still have a few spots available for our Microwave Specimen preparation workshop. The information follows. Please let me know immediately if you would like to attend. Thanks, Debby
===========================
The workshop will be held October 21, 22, and 23 in the Life Science Microscopy Facility at Purdue University, Wst Lafayette, Indiana, and run by Richard Giberson of Ted Pella, Inc.
Sunday Oct 21: introductory session starting at 2:00pm Monday Oct 22: Hands-on prep for TEM and SEM Tuesday Oct 23: Hands-on prep for ICC including block prep and ICC protocol.
We will have two microwave ovens, 2 TEM's, 1 SEM and 4 microtomes available during this time so there should be plenty of opportunity for examining the final products.
Cost for the workshop will be $350. This registration cost will include the book " Microwave Techniques and Protocols" by Giberson and Demaree, meals from dinner on Oct 21 through lunch on Oct 23, and most materials. Participants will have to provide their own resins if using ones other than Epon generic, Spurr's, or LR White. Also participants will have to provide their own specimen material (call and we will try to obtain it here if possible) and diamond knives for microtoming. Check with us if you need any other special reagents to determine if we can provide them.
The workshop will be limited to 12 participants. Let me know immediately if you would like to register and I will send info on where to send the registration checks. Please note that once you have registered the registration costs can only be refunded if we can fill the spot. All fees will be refunded if the workshop must be cancelled for any reason.
Participants will also have to provide their own transportation and hotel rooms. We have a block of rooms reserved at the University Inn. The Purdue rate is $70 for 1 person and $77/room for 2 people. We will be happy to try to match up participants who would like to share a room. Contact me for reservation information.
Those who plan to drive in can contact me for instructions. West lafayette is reached via I-65 from Indianapolis (~1 1/4 hr northwest) or Chicago (2+ hr. southeast) and via I-74 from central Illinois (~1 1/2h northwest from Champagne-Urbana).
Those who plan to fly can reach us by flying into Indianapolis and taking ground transportation to campus (~ 1 1/2 hr.. and $20 one-way). Northwest Airlines has commuter flights directly into the Lafayette/Purdue airport from Detroit. Note that we stay on Eastern Standard time all year round so are on Central time in the summer and Eastern time in the winter (at least we don't have to remember to change our clocks!). Planes presently fly into Lafayette leaving Detroit at 5:10 Saturday nights. There are only evening flights coming in on Sunday. Flights to Detroit leave at 6:01 in the evening or 7:15 in the morning during the week.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
On Wednesday, September 26, 2001 8:48 AM, Tindall, Randy D. {TindallR-at-missouri.edu} wrote: } Hi Debby, } } I'd like to receive information about this workshop. (Lou Ross forwarded } your message to me.) I doubt if I can get funding to make it, but I may } try. } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } }
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA22214 for dist-Microscopy; Thu, 4 Oct 2001 10:30:25 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA22206 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 4 Oct 2001 10:29:54 -0500 (CDT) Received: from umc-mail01.missouri.edu (umc-mail01.missouri.edu [128.206.10.216]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA22199 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 4 Oct 2001 10:29:43 -0500 (CDT) Received: from [128.206.78.175] (mu-078175.dhcp.missouri.edu [128.206.78.175]) by umc-mail01.missouri.edu with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2653.13) id 4GNN209J; Thu, 4 Oct 2001 10:26:31 -0500 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Sender: RossLM-at-pop.email.missouri.edu Message-Id: {v04011700b7e21dd57438-at-[128.206.78.175]}
Hi,
The Microbeam Analysis Society (not MSA) membership renewal packets were mailed out late last week. Inside the packet you will find a membership renewal form, a M&M Journal subscription form and a ballot for 2002 Executive Council positions. There is a deadline for the 2002 M&M Journal subscription of December 15, 2001 and the ballots must be postmarked by December 31, 2001. Everything can be returned in the envelope provided. Please take a few minutes to fill out the forms and ballot and drop them in the mail as soon as possible. This will help to avoid any future problems.
If you have not received your packet in the next few days, have any questions, or if you are not a member of the Microbeam Analysis Society and would like to join, please contact me at the email addresses or phone numbers below.
Thank you for your co-operation. Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.bioterc.missouri.edu/emc
Hello everybody Can you recommend any references that might have total emitted electron coefficient for different materials as a function of beam energy? I am interested in oxides, particularly in materials like PZT and BaTiO3. Thanks in advance Sergei
-- Sergei V. Kalinin Dept. Mat. Sci. Eng. University of Pennsylvania, 3231 Walnut St, Philadelphia PA 19104 Tel: (215) 898-3446 Fax: (215) 573-2128 E-mail: sergei2-at-seas.upenn.edu URL: http://www.seas.upenn.edu/~sergei2
A colleage of mine is inquiring if anyone knows of a person who can do routine servicing of MT2-B Porter Bloom microtomes in Central States area (central Illinois, St. Louis, Mo).
The only name they have is Bill McGee from New York. They are worried that getting someone from NY area would be very costly.
Respond directly to Dr. Chang at: Chang.Nada-at-uis.edu
OR, I will forward any responses.
Thank you.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
I made a drastic mistake in my earlier email announcement. Ballots must be postmarked by DECEMBER 1, 2001 to be counted. I apologize for the wrong date. All the more incentive to return your application, subscription and ballot as soon as possible.
Thanks, Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) www.microanalysis.org
Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.biotech.missouri.edu/emc
Food Structure & Functionality Symposium 2002 May 5 to 8, 2002, Palais des Congres de Montreal, Montreal, Quebec, Canada.
An international symposium leading Food Structure & Functionality studies through the 21st century "webaddress at the AOCS site (http://www.aocs.org/member/division/fsff/index.htm)" Held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
The mandate of the Food Structure & Functionality Forum : "To promote global collaboration between Food and Agriculture professionals in Structure and Functionality disciplines by facilitating and providing a forum for exchange of knowledge, expertise and research findings".
The symposium has two themes: * new and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Program Schedule Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl)
Monday, May 6th Opening of symposium Plenary Speaker Dairy Applications Session. Chairs: Mark Auty (mauty-at-moorepark.teagasc.ie) and Harjinder Singh (H.Singh-at-massey.ac.nz) Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl) and David Pechak (Dpechak-at-kraft.com)
Dedicated Poster Session Division Board Meeting
Tuesday, May 7th Agricultural Applications of Microscopy and Imaging Session./ joint with Feed Microscopy Division. Topic/Tentative title: New Microscopic Techniques for Identifying Food/Feed Constituents and Contaminants. contacts: Mark Auty (mauty-at-moorepark.teagasc.ie) and Kim Koch Division Luncheon and round table (expert) discussion. Topic TBA Microbiology and Food Session. Chairs Judy Arnold (jarnold-at-saa.ars.usda.gov) and Ida Yates (iyates-at-ars.usda.gov)
Division Members Meeting
Wednesday, May 8th Ingredients and Food Processing Session. Chairs: Diana Kittleson (dkittleson-at-pillsbury.com) and Bernhard Tauscher (bernhard.tauscher-at-bfe.uni-karlsruhe.de)
New Methods and Techniques for Food Structure and Functionality Analysis Session. Chairs: Kathy Groves (Kgroves-at-LFRA.co.uk) and Maud Langton (maud.langton-at-sik.se)
Closure of Symposium.
¯------------------------------------------------------------------------------------------------------------------ For more information visit the websites mentioned above, contact any of the session chairs listed above or myself.
Paula Allan Wojtas, Chair, Food Structure and Functionality Forum - a Division of AOCS
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Hi, We are trying to locate Mark Rigler of Materials Analytical Services. Have they gone out of business? Their 800# says "all circuits are busy now"...it has been like that for days. thanks for your help, Beth
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
Hi everybody, I am the operator of the SEM for my university and I am trying to shop for these two accesorries 1. Coater 2. EDX accesories The SEM is a LEO 1400. Soji
Hi All: I am looking for suggestions from experienced microscopists for voltage and current settings for the final cleaning to remove ion milling damage on Silicon and GaAs samples for High Resolution microscopy using a Gatan 600 ion mill. Presently we use the standard 5Kv, 0.5 milliamps/gun with "splotchy" and/or inconsistent results ( we are trying "tweak" our settings in order to reduce/eliminate the amorphous regions). Thanks in advance, Mike UT Arlington
The lowest you can go on the Duomill is about 10 degrees on the top side (with no top plate) and 12 degrees on the bottom side. With normal guns, you can go to about 2.5 - 3 kV with 2.5 mA.
If you insist on ion milling and you can only use the duomill, you can buy the PIPS sample holders and a pedestal from Gatan that can be used in the duomill to go to low angles. The lowest angles on dimple samples will be about 4 degrees on the dimpled side (assuming single sided dimpling). This should help you quite a bit. You will not have a lot of beam current to work with, but it should give you what you want with a little experimentation.
Now if you want no amorphous areas at all, (and I do mean none) you should use the small angle cleavage technique. Check out the Microcleave(TM) Kit from South Bay Technology on their web site. They have a couple of my PDF files that illustrate the quality of the samples that you can get.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Friday, October 05, 2001 3:47 AM To: listserver
Hi All: I am looking for suggestions from experienced microscopists for voltage and current settings for the final cleaning to remove ion milling damage on Silicon and GaAs samples for High Resolution microscopy using a Gatan 600 ion mill. Presently we use the standard 5Kv, 0.5 milliamps/gun with "splotchy" and/or inconsistent results ( we are trying "tweak" our settings in order to reduce/eliminate the amorphous regions). Thanks in advance, Mike UT Arlington
I look after a JEOL 840 with EDS and 3 x WDS in a small department.
I usually put it in standby and there's no useage while I'm away, but I'm being urged to train a deputy to mind it and help/supervise users.
But when I think about the possibility of returning to find the column and/or the spectros full of oil, or some other nightmare scenario that I (thankfully) haven't yet envisaged, I become apprehensive.
How do other places/people handle the problem of absences of the primary/sole caregiver?
thanks
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Does anyone have any information on Schiff's reagent that contains thionine or any other blue stain. If so who is the manufacturer, who is the distributor, what's the cost, what's the shelf life, etc., etc.. I am looking for an alternative Feulgen stain for DNA ploidy analysis via Image Analysis.
Thank you in advance.
Donald G. Awbrey, HT(ASCP) QIHC Image Analysis / Electron Microscopy donaldawbrey-at-texashealth.org donaldawbrey-at-hotmail.com
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, October 06, 2001 2:47 AM
Dear Listers,
I have a JEOL 35C with a defective high voltage cable. It appears to be "open" at the feedthrough to the gun. As the SEM is quite old JEOL no longer supplies replacement cables.
Morning Don, But the Feulgen Schiff has been proved to be stoichiometric! There are alternatives, but you would have to prepare them yourself and insure that the stoichiometry holds, but, then, I have always made my own Schiff reagent anyway. (Method on request, but it is readily available.) [A.G.E. Pearse, Histochemistry, Theoretical and Applied, (4th Ed.), Churchill Livingstone, New York, 1985, Vol II, ISBN: 0-443-02997-0] always has plentiful methods, and the most recent edition (Vol II) not only has many methods [including alternatives to Schiff!] but a DEEP discussion of the entire Feulgen-DNA methodology and history. Hope this helps.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Awbrey, Donald } Sent: Friday, October 5, 2001 6:41 PM } To: 'microscopy-at-msa.microscopy.com' } Subject: Schiff Reagent for Image Analysis } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hello fellow histo/micro netters, } } Does anyone have any information on Schiff's reagent that contains } thionine } or any other blue stain. If so who is the manufacturer, who is the } distributor, what's the cost, what's the shelf life, etc., etc.. I am } looking for an alternative Feulgen stain for DNA ploidy analysis via Image } Analysis. } } Thank you in advance. } } } } Donald G. Awbrey, HT(ASCP) QIHC } Image Analysis / Electron Microscopy } donaldawbrey-at-texashealth.org } donaldawbrey-at-hotmail.com } } }
We recently purchased a DVD RAM drive to assist with image storage. The problem is that our IT department and local venders don't know much about the media (The actual disks). We have two choices for DVD RAM disks: authoring or general use. Has anyone out there figured out which is the best for general microscopy images. The disks will travel between Mac's, PC's and LINUX/UNIX operating systems.
TIA,
Jon Ekman Florida State University Biological Science Imaging Resource 119 Bio Unit I, 4370 Tallahassee, FL 32306 tel: 850.644.6519 fax: 850.644.0481
John H. L. Watson Memorial Student Scholarships 15th International Congress on Electron Microscopy (ICEM-15), Durban, South Africa, Sep 1-6, 2002
In memory of John H. L. Watson, the Microscopy Society of America (MSA) has established a Travel Scholarship Award to encourage graduate students and post-doctoral research assistants who are MSA members in good standing, to attend international microscopy meetings. (Note: In the future, The John L. Watson Memorial Student Scholarships should also be available to enable young researchers from Latin America and South America to attend appropriate international meetings hosted by the MSA in the USA).
The MSA will pay from $500 to $1250 towards travel and accommodation to allow graduate students or post-doctoral research assistants (with Doctoral degrees awarded on or after January 1, 1998) who are members of the MSA to attend the 15th International Congress on Electron Microscopy (ICEM-15), in Durban, South Africa, September 1-6, 2002. The level of the monetary award will be based on the scientific content of the paper to be presented and on the number of successful applicants.
Applicants should forward to the MSA Awards Committee Chair, Dr. J. Bentley, MS-6136 4500-S, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Oak Ridge, TN 37831-6136: (i) their full name, age, address and details of academic and/or professional status, (ii) a letter of support from their advisor/supervisor that also attests to the eligibility of the applicant, and (iii) an "advanced" copy of their 2-page paper/extended abstract intended for submission to ICEM-15, on which the applicant is the lead author.
The deadline for submission is December 15, 2001. This date will allow time for evaluation of the paper by the MSA Awards Committee, confirmation with the MSA Business Office that the applicant is a member in good standing and has renewed their dues for 2002, selection/approval by MSA President, and notification of successful and unsuccessful applicants in time to meet the submission deadline of papers to ICEM-15 (February 1, 2002). Successful applicants will be required to submit an appropriate paper/extended abstract in sufficient time and of sufficient scientific content to be published in the ICEM-15 proceedings. Unsuccessful applicants will be encouraged to submit an abstract to ICEM-15, but will receive no financial support from this award.
__________________________________________________________________________ Jim Bentley Microscopy Microanalysis Microstructures Group Metals and Ceramics Division Bldg. 4500-S, MS-6136 Oak Ridge National Laboratory PO Box 2008 Oak Ridge, TN 37831-6136, USA
Tel: (865)574-5067 Fax: (865)241-3650 bentleyj-at-ornl.gov express mail use "1 Bethel Valley Rd" instead of PO Box. ** Note the new group name, mail-stop, fax, and area code **
General use media is all that you need. There are two types, Type 1 5.2GB and Type 2 2.6GB. Type 1 is double sided and can only be used in its container. Type 2 can be used in the container or removed for reading in DVD-ROM drives.
Check your drive and media carefully. I've used Panasonic and Pioneer drives, TDK and Maxell media. When writing data to the media, I had no problem. When reading the data from the media, many files were corrupted. Any file larger than about 1MB was likely to be damaged. Be sure to perform a verify after writing. Routine scandisk is also a good idea.
I don't use DVD-RAM any more since I cannot trust them. I switched to DVD-R and DVD-RW. No problems with this product.
gary g.
At 08:50 AM 10/6/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm particularly interested in an electron microscopy application, hope someone out there can help or point me in the right direction.
What I want to do is follow the route of a phagocytosed bacterium from the cell surface attaching initially to the cell surface or pseudopods to being engulfed right through to it's final destination within a lysosome and all the intervening steps in between. I have a very good antibody to a cell surface antigen on the bacteria which has been shown to work well in an immunogold labelling previously so I'd like to try and repeat this.
As I have no experience with TEM or indeed preparation of the samples I would appreciate very much any useful comments or protocols on this matter. I intend to grow the cells in flasks then add the bacteria and take a variety of time points to catch the range of stages of the bacteria through the cell - is there perhaps a better way to setup this experiment ? How might I collect the sampes, ie cell scraping or limited ezymatic digestion to get the cells of the cell culture plat ? I really want to preserve cell morphology as much as possible and the show the very early contacts between macrophage & bacterium at the cell surface so am concerned about the best way to harvest the cells, ie enzymatic harvseting or cell scraping may be worrysome. Is it possibe to process the cells for TEM & immunogold analysis whilst still attached to the plastic of a cell culture flask (ie break the flask open and somehow embed/ process the cells whilst still in-situ ?).
Any useful comments would be greatly appreciated !
In Europe when I was hopping from country to country I found that the organisations that work with X-ray sets in hospitals etc are more than capable of fixing a high voltage cable. We have even replaced the complete cable simply taking the two ends from the EM cable.
Hope this helps?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
You bring up a very good issue of removing amorphous damage from specimens which has become of increasing interest in HREM applications. Removing amorphous damage from specimens has been most recently addressed with the advent of low energy ion milling. This is a technique where the final milling stage is done at ~ 100-500eV. This is based on technology developed by Dr. Arpad Barna in Budapest and has shown some tremendous results. We market the aptly named "Gentle Mill" and you can find results for both silicon and GaAs on our website at www.southbaytech.com. Go to the search function and enter "gentle" and that will bring you to the right page. You need to register to download the PDF - if that's a bother, just send me an email and I will email the file to you.
I am sorry for the somewhat commercial nature of the response, but I do think it appropriately addresses the issue. I hope it helps.
Best regards-
David
"Walck, Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The lowest you can go on the Duomill is about 10 degrees on the top side (with no top plate) and 12 degrees on the bottom side. With normal guns, you can go to about 2.5 - 3 kV with 2.5 mA. } } If you insist on ion milling and you can only use the duomill, you can buy the PIPS sample holders and a pedestal from Gatan that can be used in the duomill to go to low angles. The lowest angles on dimple samples will be about 4 degrees on the dimpled side (assuming single sided dimpling). This should help you quite a bit. You will not have a lot of beam current to work with, but it should give you what you want with a little experimentation. } } Now if you want no amorphous areas at all, (and I do mean none) you should use the small angle cleavage technique. Check out the Microcleave(TM) Kit from South Bay Technology on their web site. They have a couple of my PDF files that illustrate the quality of the samples that you can get. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } P. O. Box 11472 (letters) } Guys Run Rd. (packages) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8515 (fax) } } -----Original Message----- } } From: coviello [mailto:coviello-at-mae.uta.edu] } Sent: Friday, October 05, 2001 3:47 AM } To: listserver } Subject: TEM-HRTEM samples with Gatan 600 ion mill } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi All: } I am looking for suggestions from experienced microscopists for voltage } and current settings for the final cleaning to remove ion milling damage } on Silicon and GaAs samples for High Resolution microscopy using a Gatan } 600 ion mill. Presently we use the standard 5Kv, 0.5 milliamps/gun with } "splotchy" and/or inconsistent results ( we are trying "tweak" our } settings in order to reduce/eliminate the amorphous regions). } Thanks in advance, } Mike } UT Arlington
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
We are using magneto-optical disks (MO-disk) for 5+ years. Since that time we had 1 disk damaged (and successfully recovered) out of few hundreds which is in circulation currently. Each disk is 2.6 or 5.2 Gb and 9-something available now. The disk uses FAT or NTFS format. DVD format is not established well and a subject of frequent changes over last few years, so I would avoid it unless they establish standard and will follow it. "FAT"- disks are readable from any platform and no software necessary to operate the drive (it's SCSII) - it's visible on desktop immediately after connection.
Sergey
At 01:29 PM 10/6/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Dear All, We recently purchased a new Leica EM Stainer to replace our aging (but otherwise pretty trusty) LKB Ultrostainer and have some misgivings about our new acquisition. Until the company has had a bit longer to try to sort the problems I do not want to go into specifics here but despite of spending several months trying to sort out theses 'issues' we are a long way from being happy with the instrument - this is despite it having been back to the factory once.
I would really like to know if other owners and users of the new EM Stain have experienced major problems with this piece of equipment - or if its just our one (or us!). If your EM Stain has proved totally satisfactory, I'd like to hear from you too...
Best regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
Nextek Inc. is seeking a qualified individual to join our analytical laboratory team as a Materials/Failure Analysis Engineer. The ideal candidate will have a B.S. in Materials Science, Chemistry, Engineering or equivalent, and a minimum 3 years experience in the micro-electronic field; or 5 or more years hands-on lab experience in the micro-electronic materials/failure analysis field. Knowledge in cross-sectional and destructive analysis of micro-electronic assemblies and devices, and experience with analytical instrumentation such as; SEM/EDS, Micro-FTIR, Thermal Analysis, Acoustic and Optical Microscopy is a plus.
To learn more about Nextek Inc. visit our web site at http://www.nextekinc.com/. For more information about this position, call or email the following:
Wade McFaddin (256) 772-1995 ext.1064 wade.mcfaddin-at-nextekinc.com or Jim Chiang (256) 772-1995 ext. 1029 james.chiang-at-nextekinc.com
Dear All: I am looking for a monoclonal anti pancreatic amylase antibody. Does anyone out there know a source. Thank you in advance.
Hong ------------ Hong Yi Emory Neurology Microscopy Core Laboratory Emory University Department of Neurology 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30322 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
Many years ago, when (or even before) I was an undergraduate (that would place it in the early '60s), my aunt managed the analytical lab at a company that produced non-ferrous metals (principally, I think, bronzes and brasses). As one does in such situations, she invited me to tour her labs one day.
One of the instruments I was shown was called a "Beta Probe". A piece of the metal was ground and polished, and was then placed face down on an aperture in the top plate, with what I now realize was an O-ring seal. A few buttons were pressed, and after a little while some of the electronic counters of the day (neon tubes with 10 electrodes) began spinning round. My memory is that the operator had some range of ratios between the recorded numbers within which he (it was, of course, a "he" in those days, even though my aunt was the manager) could pass the sample as meeting specifications.
I surmise that this was an electron milli-probe (if you'll forgive my coining of the term!) with preset spectrometers tuned to the elements of interest in the specification. I would very much like to find out more about this device. Happily, my aunt is still available to be asked, but she knows nothing of the operation of the instrument. The people who ran it have either died, or lost touch (the company failed twenty years ago).
If anyone can give me more information about this instrument, I would be very interested, and most grateful.
The University of New Hampshire is looking for a Director of the Instrumentation Center. For more information please go to our website at:
www.unh.edu/instrumentation-center/index
Analytical services at the UIC include: Elemental Analysis; Gas Chromatography-Mass Spectrometry (GC-MS); High-Field Nuclear Magnetic Resonance (FTNMR - 90, 360, 400, and 500 MHz); Ultra-Violet, Visible, and Near-Infrared Spectrophotometry (UV-Vis-NIR); Infrared Spectroscopy (FTIR); Optical Rotation; X-Ray Photoelectron Spectroscopy; Electron Microscopy including Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) with Energy Dispersive Spectroscopy (EDS).
RESPONSIBILITIES: Reporting to the Vice President for Research and Public Service, the University Instrumentation Center Director will be responsible for leading an emerging research and teaching oriented service center toward excellence; duties to include:
Working directly with faculty, administrators, and clients from diverse backgrounds and perspectives to maximize use of all facilities within the UIC; forging partnerships with researchers, teaching oriented faculty members, other departments, programs, and research centers and institutes for purposes of proactively supporting research and instruction;
Managing the financial, administrative, and supervisory aspects of the UIC;
Supporting education and training of staff, university researchers/clients, and students;
Securing funding through external and internal sources in order to obtain new instruments and upgrade existing equipment;
Promoting UNH outreach mission by developing knowledge and awareness of other laboratories in the state and region, for purposes of forming partnerships, referral relationships, information networks, etc. Serving as a resource for state government and business and industry.
QUALIFICATIONS:
Advanced degree in physical sciences or engineering.
Minimum of three years managerial experience.
Demonstrated experience with operation of analytical instruments and application to research problems.
Familiarity with securing external funding for scientific instruments.
Excellent oral and written communication skills.
Demonstrated ability to work with a variety of constituents.
STARTING DATE: Negotiable, as early as 12-1-01
SALARY: Competitive and commensurate with experience and local area.
APPLICATION DEADLINE: Open until filled.
APPLICATION PROCEDURE: Applicants should send a letter of intent, CV/resume, and the names, addresses, and phone numbers of three references to:
Chair, University Instrumentation Center Director Search Committee
C/o Office of the Vice President for Research and Public Service
Room 107, Thompson Hall
University of New Hampshire
Durham, NH 03824
Electronic resumes and supporting documentation will be accepted. Please e-mail your package to: diana.couture-at-unh.edu
Can anyone offer any ideas on the best way to proceed with removing the effects of charging on an image? I am referring to the streak across the image manifested as a change in density. As an example, have a look at this uncoated mite:
http://katie.ucdavis.edu/pics/sucker2.jpg
The nature of this projects requires that we place the live mite in the scope and get a picture in the first 5 minutes. This picture was taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor for publication. I am looking for some help in eliminating some of the charging artifacts without losing too much detail. Any help will be appreciated.
We are using PS 6 and I have the IPTK plugins. I made some progress using a Gaussian filter to produce a very fuzzy duplicate image, invert the image, add it to a layer and then adjust the opacity to around 50%. This was only partially successful. Any help will be appreciated.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
I would be interested to hear of any IP solution you come up with.
However, I would think that running the scope in environmental mode at higher voltage ought to be able to give you a satisfactory image.
We have a Hitachi S2460N here. We routinely run 40 Pa of Helium to eliminate charging. At more than 10 kV, we can get a suitable image off of our Robinson BSE. Our Oxford Tetra(tm) allows us to drop the voltage further. I will forward you an image of a bug from our scope under separate cover.
Warren
At 09:29 AM 10/9/2001 -0700, you wrote:
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
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In a message dated 10/9/01 12:45:01 PM, raharris-at-ucdavis.edu writes:
} We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated.
Another approach you might try is to duplicate the image, smooth the copy with a Gaussian blur, and then divide by it. The size of the blur to use depends on the size of the bright streaks, and these are not consistent in your image, but values in the range of 2-4 pixels seem to work fairly well.
Hi Rick, If the image is saturated with white pixels ("blown out highlights") then your are pretty stuck. There isn't anything that can be done to put information in where it doesn't exist. However, in order to make the most out of a bad situation, you can use the levels and curves options to adjust the brightness and contrast for a reasonable image. I prefer to use adjustment layers to do this, that way I can edit and tweak the settings without permanently changing the core data. Your best best is to avoid/minimize saturation when you capture your images. A "wet" ESEM is fantastic for this. As you probably know, try to minimize spot size, current, and voltage to minimize charging artifacts, and choose brightness and contrast settings that don't saturate the image. If you want to see what I mean by adjustment layers using curves and levels, you can send me a file, and I'll set it up for you, and send it back for you to look at. Good luck.
-Brad
---------- From: Rick Harris Sent: Tuesday, October 9, 2001 9:29 AM To: Microscopy-at-sparc5.microscopy.com Subject: Photoshop and SEM charging
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Can anyone offer any ideas on the best way to proceed with removing the effects of charging on an image? I am referring to the streak across the image manifested as a change in density. As an example, have a look at this uncoated mite:
http://katie.ucdavis.edu/pics/sucker2.jpg
The nature of this projects requires that we place the live mite in the scope and get a picture in the first 5 minutes. This picture was taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor
for publication. I am looking for some help in eliminating some of the charging artifacts without losing too much detail. Any help will be appreciated.
We are using PS 6 and I have the IPTK plugins. I made some progress using a Gaussian filter to produce a very fuzzy duplicate image, invert the image, add it to a layer and then adjust the opacity to around 50%. This was only partially successful. Any help will be appreciated.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Rick, My experience is that live insects don't generally charge because the evaporating water carries the charge away. From the looks of the image, I think you were using a lot of beam current. Cur that back a whole lot and the charging will probably go away. Also, the horizontal scan speed has a much greater effect on charging than the vertical speed. If you run a faster horizontal rate and more lines (slower vertical speed), this will help.
Ken Converse owner Quality Images third party SEM service Delta, PA
Rick Harris wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone offer any ideas on the best way to proceed with removing } the effects of charging on an image? I am referring to the streak } across the image manifested as a change in density. As an example, } have a look at this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in } the scope and get a picture in the first 5 minutes. This picture was } taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but } is too poor for publication. I am looking for some help in } eliminating some of the charging artifacts without losing too much } detail. Any help will be appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress } using a Gaussian filter to produce a very fuzzy duplicate image, } invert the image, add it to a layer and then adjust the opacity to } around 50%. This was only partially successful. Any help will be } appreciated. } } } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu } } } }
Our facility is looking to purchase a dry 1x objective for a Nikon Microphot light microscope. I have contacted a few vendors and have been told that the microscope is out of date and it will be impossible to locate such an objective. Any help in locating this item will be appreciated.
Twin City International Inc. made a beta backscatter instrument called the Betascope. It used an isotope source and a Geiger-Mueller tube to measure the thickness of metal films after a simple calibration with known thickness standards. I don't know if they are still in business.
-----Original Message----- } From: Tony Garratt-Reed [mailto:tonygr-at-mit.edu] Sent: Tuesday, October 09, 2001 7:59 AM To: microscopy-at-sparc5.microscopy.com
Hi, Listers!
Many years ago, when (or even before) I was an undergraduate (that would place it in the early '60s), my aunt managed the analytical lab at a company that produced non-ferrous metals (principally, I think, bronzes and brasses). As one does in such situations, she invited me to tour her labs one day.
One of the instruments I was shown was called a "Beta Probe". A piece of the metal was ground and polished, and was then placed face down on an aperture in the top plate, with what I now realize was an O-ring seal. A few buttons were pressed, and after a little while some of the electronic counters of the day (neon tubes with 10 electrodes) began spinning round. My memory is that the operator had some range of ratios between the recorded numbers within which he (it was, of course, a "he" in those days, even though my aunt was the manager) could pass the sample as meeting specifications.
I surmise that this was an electron milli-probe (if you'll forgive my coining of the term!) with preset spectrometers tuned to the elements of interest in the specification. I would very much like to find out more about this device. Happily, my aunt is still available to be asked, but she knows nothing of the operation of the instrument. The people who ran it have either died, or lost touch (the company failed twenty years ago).
If anyone can give me more information about this instrument, I would be very interested, and most grateful.
Does anyone know of a dye that binds to cellulose (or pectins) and is fluorescent? I am looking for an alternative to calcofluor. I wish to stain 1 micron thick LR-white sections of Vicia faba cotyledons.
Hi there, I know this question has been asked lots, but i never needed to pay any attention till now, so my apologies. My institute wishes to purchase a very good digital camera for light / fluorescence microscopy to replace (compliment?) the 35 mm film cameras that we currently use. Part of the requirements is that the camera be able to be taken off the scope and have a regular lens fitted for other high / professional quality images (such as whole plants, animals etc.) So the question I have to ask is, is this a reasonable task for a system? Further to that, what do current users of digital cameras for microscopy see as an absolute requirement and what is nice to have?
A short list of cameras suggested to us locally are: Canon D30 3.3 megapixel, Fuji/Nikon S1 3.3mp, Nikon DiX 5.5 mp. Any comments on those (good or bad)? Any other recommendations?
John Blinco Western Australian State Agricultural Biotechnology Center
Mark Congo red would be a suitable alternative. Excite with green and view in orange/red - Rhodamine -type filter set.
You could also try a fluorescent pseudo-schiff reaction using periodic acid or sodium m-periodate oxidation followed by staining with a carbohydrazide derivative of a fluorescent dye - e.g. Lucifer Yellow CH.
A dye suitable for uronic acid polysaccharides such as pectin is Coriphosphine O CI No. 46020. Again this works with Rhodamine type filter sets
Chris
Date sent: Wed, 10 Oct 2001 12:21:11 +1000 } From: MARK JEFFREY TALBOT {mark.talbot-at-studentmail.newcastle.edu.au}
The Hitachi S3500N has the N-SEM mode which should give you a reasonable image in backscatter, with appropriate WD and pressure settings. Also the 3500 has an integrate capture mode to do multiple rapid scans which can help reduce charging.
David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/micros_e.html
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: October 9, 2001 1:30 PM To: Microscopy-at-sparc5.microscopy.com
Can anyone offer any ideas on the best way to proceed with removing the effects of charging on an image? I am referring to the streak across the image manifested as a change in density. As an example, have a look at this uncoated mite:
http://katie.ucdavis.edu/pics/sucker2.jpg
The nature of this projects requires that we place the live mite in the scope and get a picture in the first 5 minutes. This picture was taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor for publication. I am looking for some help in eliminating some of the charging artifacts without losing too much detail. Any help will be appreciated.
We are using PS 6 and I have the IPTK plugins. I made some progress using a Gaussian filter to produce a very fuzzy duplicate image, invert the image, add it to a layer and then adjust the opacity to around 50%. This was only partially successful. Any help will be appreciated.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
One of those answering Rick Harris' question on charging remarked about using helium in the variable pressure mode. What other gasses are used for the VPSEM and ESEM? What considerations are taken into acccount in choosing a vent gas? Is there a paper somewhere that answers these questions?
We use Congo Red and image it in the rhodamine channel...
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One of our pharmacology researchers is making nanoparticles ranging in sizes from 50nm up to 250nm. The material is polylactic-acid-coglycolic-acid (PLGA). He's looking for a lipophilic electron dense particle, dye or stain that could be incorporated into the particles during their formation and which could be seen in the TEM sections. The material has to be able to be suspended or dissolved in an organic solvent.
For our part here are some of the things we've thought of using: Ruthenium Red powder, colloidal gold in 5 to 10nm range, phosphotungstic acid in ethanol.
His material will also melt at 45 degrees centigrade. So I embedded in Unicryl, UV polymerization at 4 degrees centigrade. Does anyone know if there is still heat produced during the polymerization?
This seems to be in the realm of polymer chemistry and materials science. I'd appreciate any and all help, Thanks.
We run a Hitachi S2250N. Rick, what gas, pressure and WD were you using? Helium is the best, as Warren says, but if you dont have a tank handy, water vapour (for a minimal system - just connect the gas inlet to a half-full flask of water!) is almost as good for imaging. And I would also suggest a higher voltage - 5kV minumum, 10-15 for comfort, if you want to work quickly at low magnifications and can afford to miss a bit of surface detail. On something with as many projections as a mite, utilising the VP system to eliminate charging will let you work much more rapidly than relying on low kV alone. Its not very PC, but we would tend to start at 20kV and work down....
regards Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
} } } Warren E Straszheim {wesaia-at-iastate.edu} 10/10/01 03:30AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would be interested to hear of any IP solution you come up with.
However, I would think that running the scope in environmental mode at higher voltage ought to be able to give you a satisfactory image.
We have a Hitachi S2460N here. We routinely run 40 Pa of Helium to eliminate charging. At more than 10 kV, we can get a suitable image off of our Robinson BSE. Our Oxford Tetra(tm) allows us to drop the voltage further. I will forward you an image of a bug from our scope under separate cover.
Warren
At 09:29 AM 10/9/2001 -0700, you wrote:
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I'm not an expert at this, but I do play with images in photoshop for my own amusement.
I played with your image in Photoshop and the best I could do was reduce the difference in darkness between the top and the bottom of the picture by lasooing the top portion of the picture (at 200X mag.) with the lasoo tool and then using the brightness-contrast setting under Image, Adjust, to get the top and bottom portions of the photo to match in brightness. This still left the lines, which I tried the gausian blurr on, but it couldn't be blurred much before it looked out of place. (like a blurry band in the middle of an otherwise crisp the picture.)
This is just another trick you might try. Good luck.
Karen Pawlowski
Rick Harris wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu
} Dear Listers, } } Thank you to all who have responded to my request for high voltage cable repair. } } The results were: } } 1. Fix it yourself. } 2. Buy another JEOL 35C for the cable. } 3. Have a third party repair the cable. } } The consensus is that the best third party repair is: } } } Dielectric Sciences Inc. } } 88 Turnpike Road } } Chelmsford, MA 01824 } } Phone: 978 250-1507, FAX: 978 250-1699 } } } Best Regards, } } Earl Weltmer
We need some help for solving a problem related to the deformation of the image taken with our SEM . We have already asked a number of people about this but, it seems the solution is not so straightforward.
We have a JEOL SEM 35C, from the early 804s. Recently, we installed the SemAfore digital acquisition system which gets the electronic signal directly out of the column and sends it to the computer. The CRT4s show the same analogic image as always.
The distorsion of the image appears at the beginning of every scanline ramp (lefthand side on the image). Two micrographs digitally taken from latex microspheres give the idea what the problem looks like: the same feature (a group of microspheres) is taken positioning them at the very left-hand side of the image, then moved to the right-hand side of the same CRT screen. It is clear that the distortion on the shape of the spheres appears on the first image (on the second one, you see a very very little deformation to the opposite direction). Find both micrographs at:
http://www.ceride.gov.ar/servicios/sem/kuqui1.jpg
http://www.ceride.gov.ar/servicios/sem/kuqui2.jpg
The technicians at our institute tested with the oscilloscope the shape of the signal out of the scan generator unit. See, at:
http://www.ceride.gov.ar/servicios/sem/ the file: dibujo_mail.bmp
a drawing is seen: in blue the theoreticall scan generator ramp, while in magenta the real one as shown on the oscilloscope. The scanning ramp showes a distortion on the shape starting from the begining up to the 8% of the leght of the ramp, along a scanline (see the magenta dip inside the green square). From that point, the ramp followes the shape that should have up to the end of the scanline. Rounded corners at the beginning and the end of the ramp are commonly found, according to the technicians.
This problem was seen on the CRT screen since long ago, but since the image on the CRT is diminished in size, the defect is not so obvious nor so serious. The digital imaging system gets and showes on the PC screen the complete image, made up by the complete scan along a line and, along the frame. In these conditions, the distortion shown on the digital images is dramatical.
The technicians say that a sort of over scan made via software, once the image is collected, could help. But, that solution reduces the side of the viewing field and takes extra imaging processing work.
Any suggestion or help will be very much appreciated, thanks in advance.
Silvia Montoro- Nora Pratta Centro Regional de Investigaciones y Desarrollo de Santa Fe (CERIDE) G|emes 3450 3000 Santa Fe Argentina csedax-at-ceride.gov.ar ---------------------------------------------------------------------------- ----
I have unearthed 2 EM film casettes (the boxes that store film in a TEM). They have no identifying marks. They are black, about 5 1/2 x 4 3/8 x 2 1/4 inches. The black metal top that slides out is about 5 1/2 x } 6 7/8 inches. They do not belong to Philips or JEOL microscopes, but I cannot determine where they originated, perhaps Zeiss? or Hitachi? If anyone thinks (s)he can use them, please reply directly, and I will send them to you. Otherwise, they will have to take up some space in the landfill.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Your effect looks like there is not enough fly-back delay for your SEM under your present operating conditions. We can see the same foreshortening and shifting on the monitor of our JEOL 840A as we switch through the various scan speeds. The effect is particularly pronounced at TV-rate scans. It is present, but not so bad, at the SR (super-rapid?) scan rate, and it is not apparent at the Slow-1 or Slow-2 scan settings.
I am not familiar with the SemAfore system. Is is an active system (takes control of the SEM scan) or a passive system (just digitizes the regular SEM scan)? If it is an active system, there should be an adjustment to increase the fly-back delay before the next horizontal scan starts. If they don't provide such an adjustment, they should add one. If it is a passive system, then you should slow down your SEM's scan.
Warren
At 12:27 PM 10/10/2001 -0300, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Dear colleagues, I don't know if this thread has already come up, but here goes: Does anyone have tips on using EM to identify transplanted cultured cells labelled with latex beads in a host animal. Especially to see if synapses are being formed. Would traditional methods destroy the latex? if so, are there other labels for the cultured cells? Thanks in advance, Julie Gross Dept. of Neuroscience UCONN Health Center Farmington, CT 06030
1. Brad Johnson wrote: "Your best bet is to avoid/minimize saturation when you capture your images A "wet" ESEM is fantastic for this. As you probably know, try to minimize spot size, current, and voltage to minimize charging artifacts..."
Your S3500N is well equiped to do this. Consistent with the above suggestions, have you also been playing around with the mites in the variable pressure mode? In VP mode, use your backscattered electron image which inherently shows much less charging artefact thae SE imaging, and by adjusting the pressure you may be able to eliminate charging. In BSE mode, you may need higher kV, like 5-10 kV, to "see" through the gas in the VP mode, and in order for BSE's to be detected by your BSE detector.
If you have the ESED mode on your S3500N, that will enable you to to get a SE-like image in VP mode. Still, at the low mags you are working at, BSE should do fine for sufficent resolution, and may be easier to get rid of charging in that mode.
You may even be able to succeed using BSE image in high vacuum mode, if the mite doesn't dry out too fast..
2. Ken Converse wrote: "Also, the horizontal scan speed has a much greater effect on charging than the vertical speed. If you run a faster horizontal rate and more lines (slower vertical speed), this will help."
Again on your S3500N, you have that Image Integration mode that often can be used to eliminate charging in whatever scope mode you are operating in. Select the slowest line scan rate such that you observe no charging bands on your monitor. If you still see some bands, select next fastest rate. Then go to Image Integration set-up and select the number of frames to add tgether, each recorded at the scan rate you just set, usually something like 4-10 will work.
Hope this helps, and good luck. Let us all know what works for you.
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image?...snip!...
} The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated.
} } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu
} This problem was seen on the CRT screen since long ago, but since the image } on the CRT is diminished in size, the defect is not } so obvious nor so serious. The digital imaging system gets and showes on the } PC screen the complete image, made up by the complete scan along a line and, } along the frame. In these conditions, the distortion shown on the digital } images is dramatical.
I think your clue is in the statement above. If the defect can be seen on the SEM's CRT screen, no mater how small, then your SEM has a scan generator problem. SemaAfore (a JEOL product) is a passive scan digitizer. It can only do as good as the 35C's scan generator. Your 35C's scan generator is producing ramps that are non-linear at the start and end of the ramp. Like the tech said, thats not abnormal.
What's missing from your ramp diagram is where does the SEM starts using the ramp signal for image display. What happens is the ramp is non-linear at the start but the SEM has a little delay after the start to get into the linear range before using the ramp for image display. The horizontal blanking signal defines this region.
You need to look at the horizontal blanking signal to see the real start/end of ramp location. If you are still non-linear in that ramp region, then the tech needs to fix this first on the SEM.
SemAfore might have a software param to add a delay after the horizontal blanking signal. This might fix the acquired image but you are better off fixing the SEM first.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Dear list member, I am looking for an old paper, Th. Koller, et al., Micron 1:110, (1969). This journal is not available in Hopkins. Does anyone would lile to help me? thanks
Chen Chen
Department of Biological Chemistry The Johns Hopkins University School of Medicine 725 N. Wolfe Street Baltimore, Maryland 21218
Rick, I am assuming you want to fix this image rather than go collect another one. It depends on how much time and effort you wish to put into this project. One way, that I have used rather successfully has been a combination of the clone tool with a small brush size and just a little feathering of the edge, brightess/contrast adjustment, and a little spot blurring. Do each change as a separate layer leaving your original intact. It will take some time but I can show you before and after images (much worse than yours since molds tend to charge like heck) that you'd never know where charging.
Dr.Tina S. Schwach, President Microscopy Consulting Services Inc. 651-681-0112
Hello Silvia, We recently had an EDS mapping and imaging upgrade done to our SEM. When the technician installed the imaging hardware and software, there were several adjustments made (using software settings) to select the field of view that was digitally captured. As I watched the field engineer work, it was apparent that the area selected for image capture was smaller than the scanned area. Roughly speaking, it appeared that the scanned area was about 10-15% larger than what was captured. Possibly you could make some adjustments to the image capture area such that your digital image field of view would be the same as your analog, while retaining a larger scanned area. Since every manufacturer has their own way to accomplish these things, you would probably need to get the specifics directly from them. Good luck.
-Brad
---------- From: Nora Pratta- Silvia Montoro Sent: Wednesday, October 10, 2001 8:27 AM To: microscopy-at-sparc5.microscopy.com Cc: csedax-at-ceride.gov.ar Subject: SEM: distortion on digital images
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Dear members of the list,
We need some help for solving a problem related to the deformation of the image taken with our SEM . We have already asked a number of people about this but, it seems the solution is not so straightforward.
We have a JEOL SEM 35C, from the early 804s. Recently, we installed the SemAfore digital acquisition system which gets the electronic signal directly out of the column and sends it to the computer. The CRT4s show the same analogic image as always.
The distorsion of the image appears at the beginning of every scanline ramp (lefthand side on the image). Two micrographs digitally taken from latex microspheres give the idea what the problem looks like: the same feature (a group of microspheres) is taken positioning them at the very left-hand side of the image, then moved to the right-hand side of the same CRT screen. It is clear that the distortion on the shape of the spheres appears on the first image (on the second one, you see a very very little deformation to the opposite direction). Find both micrographs at:
http://www.ceride.gov.ar/servicios/sem/kuqui1.jpg
http://www.ceride.gov.ar/servicios/sem/kuqui2.jpg
The technicians at our institute tested with the oscilloscope the shape of the signal out of the scan generator unit. See, at:
http://www.ceride.gov.ar/servicios/sem/ the file: dibujo_mail.bmp
a drawing is seen: in blue the theoreticall scan generator ramp, while in magenta the real one as shown on the oscilloscope. The scanning ramp showes a distortion on the shape starting from the begining up to the 8% of the leght of the ramp, along a scanline (see the magenta dip inside the green square). From that point, the ramp followes the shape that should have up to the end of the scanline. Rounded corners at the beginning and the end of the ramp are commonly found, according to the technicians.
This problem was seen on the CRT screen since long ago, but since the image on the CRT is diminished in size, the defect is not so obvious nor so serious. The digital imaging system gets and showes on the PC screen the complete image, made up by the complete scan along a line and, along the frame. In these conditions, the distortion shown on the digital images is dramatical.
The technicians say that a sort of over scan made via software, once the image is collected, could help. But, that solution reduces the side of the viewing field and takes extra imaging processing work.
Any suggestion or help will be very much appreciated, thanks in advance.
Silvia Montoro- Nora Pratta Centro Regional de Investigaciones y Desarrollo de Santa Fe (CERIDE) G|emes 3450 3000 Santa Fe Argentina csedax-at-ceride.gov.ar
Dear List, It seems the meter, model 505, for my Edwards penning gauge has bit the dust. Edwards no longer sells nor works on the 505's so getting it repaired doesn't seem to be an option. Would anybody perhaps have one of these "obsolete" meters gathering dust on their shelf? I would appreciate hearing from you. Thanks. Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
I just heard about Two-Photon (or Multi-Photon) excitation microscopy. Does anybody know of a web site that has basic information about this technique, descriptions of applications, and the advantages. Something like "Two-Photon Microscopy for Dummies" would do.
This has always been such a terrific source of information.
Has anyone used the anti-SKI antibody from Santa Cruz? Or any other commercial antibodies to SKI (and/or SKIP)?
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
The distortion is caused by the scan coils "ringing".
When an SEM (or imaging system) acquires an image, it rasters from left to right then restarts at the left side of the screen again.
It is the "retrace", from right to left, at a relatively high speed that causes the scan coils to generate "back emf" or "ringing".
SEM manufacturers usually "blank" ,or turn off, the video during the retrace and part of the first few milliseconds of video. The effect is that you lose part of the image that is distorted.
What can be done to resolve your problem? Slower horizontal scan speed would help. Better yet, ask the vendor to blank the image just as the SEM manufacturers do.
Hope this helps,
Earl
----- Original Message ----- } From: "Johnson, Bradley R" {Bradley.Johnson-at-pnl.gov} To: {microscopy-at-sparc5.microscopy.com} ; "'Nora Pratta- Silvia Montoro'" {csedax-at-ceride.gov.ar} Sent: Wednesday, October 10, 2001 12:59 PM
I would appreciate any information about resins that have been found best for embedding of tissues for confocal microscopy. Of particular interest are lack of autofluorescence and quenching. Thank you Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
Brad, I'm not familiar with the S3500N, but most SEMs have a means of setting your record scan speed and this usually involves up to 3 different settings: 1.) Number of lines per frame (vertical rate), 2.) number of points per line (part of horizontal rate) and 3.) dwell time per point (the other part of horizontal rate) Sometimes the last two are combined in a single adjustment of time per line.
This adjustment is usually different from the viewing scan rate adjustment.
Perhaps someone who is familiar with this SEM can tell you how to make the adjustments
Ken Converse owner Quality Images third party SEM service Delta, PA
Johnson, Bradley R wrote:
} HI Ken, } I read your message, and the comment about adjusting scan speed } interests me. How would one do that? } } -Brad } } } ---------- } } From: Ken Converse } } Sent: Tuesday, October 9, 2001 3:54 PM } } To: Rick Harris; MSA, listserver } } Subject: Re: Photoshop and SEM charging } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Rick, } } My experience is that live insects don't generally charge because the } } evaporating water carries the charge away. From the looks of the image, } } I think you were using a lot of beam current. Cur that back a whole lot } } and the charging will probably go away. Also, the horizontal scan } } speed has a much greater effect on charging than the vertical speed. If } } you run a faster horizontal rate and more lines (slower vertical speed), } } this will help. } } } } Ken Converse } } owner } } Quality Images } } third party SEM service } } Delta, PA } } } } Rick Harris wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Can anyone offer any ideas on the best way to proceed with removing } } } the effects of charging on an image? I am referring to the streak } } } across the image manifested as a change in density. As an example, } } } have a look at this uncoated mite: } } } } } } http://katie.ucdavis.edu/pics/sucker2.jpg } } } } } } The nature of this projects requires that we place the live mite in } } } the scope and get a picture in the first 5 minutes. This picture was } } } taken on a Hitachi S3500N at 2.5 KV. The picture suits our needs but } } } is too poor for publication. I am looking for some help in } } } eliminating some of the charging artifacts without losing too much } } } detail. Any help will be appreciated. } } } } } } We are using PS 6 and I have the IPTK plugins. I made some progress } } } using a Gaussian filter to produce a very fuzzy duplicate image, } } } invert the image, add it to a layer and then adjust the opacity to } } } around 50%. This was only partially successful. Any help will be } } } appreciated. } } } } } } } } } } } } Rick A. Harris, Director } } } Microscopy and Imaging Facility } } } Section of Molecular and Cellular Biology } } } 1241 Life Sciences Addition } } } University of California } } } Davis, CA } } } 530 752 2914 } } } 530 754 7536 fax } } } http://katie.ucdavis.edu } } } raharris-at-ucdavis.edu } } } } } } } } } } } }
The distortion you have looks like a problem with the syncronicity of the x-y scan control and beam blanking. As the scan moves across the specimen, the beam physically moves. The electronics of the microscope "blank" the signal as the beam drops down one scan line and returns to the left hand side of the micrograph. If the blanking mechanism is slightly out of sync with the scan coils, the signal turns on while the beam is still moving from left to right. The distortion is alway at the left hand side of the micrograph and appears as a mirror image on the left and right sides of a blur. It usually is worse at faster scan rates. You may be able to get rid of the distortion by using a slower scan speed.
Cold Spring Harbor Laboratory on the north shore of Long Island, New York is seeking an experienced and responsible Biological Microscopy Facility Core Manager for the laboratory's state-of-the-art central microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, and digital imaging. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org -- Dr. David L. Spector Cold Spring Harbor Laboratory One Bungtown Road Cold Spring Harbor, New York 11724 Tel. (516) 367-8456 Fax (516) 367-8876 email: spector-at-cshl.org
Greetings Darrell, There are some good resources for learning more about multiphoton microscopy on the web. Gaining a solid conceptual understanding of the theory requires learning a little (and later, a lot) about the lasers used in this modality of optical sectioning microscopy. Here are some of the best resources I've come across:
http://swehsc.pharmacy.arizona.edu/exppath/micro/confocal.html (a great starting point for links to optical sectioning microscopy)
http://microscopy.fsu.edu/primer/techniques/fluorescence/multiphoton/multiphotonhome.html (has excellent animated renditions of the concepts)
http://www.coherentinc.com/cohrLasersAPPLICATIONS/assets/applets/CLGMPE.pdf (a PDF document from coherent, inc., a provider of lasers for multiphoton microscopy)
and you may find some helpful information and links on our site (I'm presently putting together some pertinent instructional material for our website):
http://www.itg.uiuc.edu/
If you have any specific questions, I'll be happy to try and answer them for you.
-Karl G.
_______________________________________________ Karl Garsha Specialist in Light Microscopy Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
This thread on Photoshop "enhancement" of electronic raw date brings to light an area of constant concern within my laboratory. At what point have we over "enhanced" the image, creating/altering features which potentially do not exist within the sample? It seems clear that any attempt to clone, paint, or copy/ paste fall into the category of digital artistry and should be avoid. In fact, most of the respondents have attempted to address the reduction/elimination of sample charging within the SEM, thus eliminating the need for image manipulation. Due to the highly regulated environment of the pharmaceutics industry we must adhere to strict FDA guidelines so our interpretations tend to be on the extreme side. That being said, my recommendation is to be very careful with the use of photoshop, it is a great product for "manipulating" images but our data should not be subject to artistic license. Switching directions a bit, we do employ filtering routines when analyzing data. We attempt to insert appropriate controls into the method to catch any unanticipated events. I would be very interested in the opinions of others in regards to where they draw the line on image manipulation and how they rationalize their position.
Thank you, jr
} -----Original Message----- } From: Rick Harris [SMTP:raharris-at-ucdavis.edu] } Sent: Tuesday, October 09, 2001 12:30 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Photoshop and SEM charging } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } } http://katie.ucdavis.edu/pics/sucker2.jpg } } The nature of this projects requires that we place the live mite in the } scope and get a picture in the first 5 minutes. This picture was taken on } } a Hitachi S3500N at 2.5 KV. The picture suits our needs but is too poor } for publication. I am looking for some help in eliminating some of the } charging artifacts without losing too much detail. Any help will be } appreciated. } } We are using PS 6 and I have the IPTK plugins. I made some progress using } } a Gaussian filter to produce a very fuzzy duplicate image, invert the } image, add it to a layer and then adjust the opacity to around 50%. This } was only partially successful. Any help will be appreciated. } } } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } 530 754 7536 fax } http://katie.ucdavis.edu } raharris-at-ucdavis.edu }
I have a Nikon Labophot II and was recently told that 2.5X was the lowest mag objective that was made. However, I don't know anything about the Microphot. You can post a "want to buy" add for free on labx.com. I had a very good experience doing that a short time ago. You can also do an internet search for used microscopes. You will get a lot of hits. I have found many of the used equipment dealers to be very nice and helpful.
Judy Bowen
P.S. I think you are located at SIU Springfield. I was there from 1987-89. I was Judy Rapp then and did a little EM work. I was a friend of Donna's. Send me an e-mail at jabowen-at-buckman.com if any of this is familiar.
} } Hi, } } Our facility is looking to purchase a dry 1x objective for a Nikon Microphot } light microscope. I have contacted a few vendors and have been told that } the microscope is out of date and it will be impossible to locate such an } objective. Any help in locating this item will be appreciated. } } Thank you in advance for your help } } aruna }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donsheri695-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, October 11, 2001 at 15:06:46 ---------------------------------------------------------------------------
I assume that a good soluble organometal is easier to incorporate than a suspension of powder etc, esp for such small particles (the ruthenium red powder may be larger than your nanoparticles?), but I could easily be wrong. Sounds like you don't want to just react the stain with the particle surface before doing the study, right?
What about tetraethyl lead? Check into the catalysts (e.g. stannous octoate) used to polymerize the plga or other polymers.
I'm interested, and I would appreciate it if you could let me know what is decided upon.
Richard
Richard Thrift Richard_Thrift-at-SkyePharma.com SkyePharma, Inc 10450 Science Center Drive San Diego, CA, 92121 USA
} } } {"tbargar-at-unmc.edu"-at-sparc5.microscopy.com} 10/10/01 6:19:05 AM } } } Hi everyone,
One of our pharmacology researchers is making nanoparticles ranging in sizes from 50nm up to 250nm. The material is polylactic-acid-coglycolic-acid (PLGA). He's looking for a lipophilic electron dense particle, dye or stain that could be incorporated into the particles during their formation and which could be seen in the TEM sections. The material has to be able to be suspended or dissolved in an organic solvent.
For our part here are some of the things we've thought of using: Ruthenium Red powder, colloidal gold in 5 to 10nm range, phosphotungstic acid in ethanol.
His material will also melt at 45 degrees centigrade. So I embedded in Unicryl, UV polymerization at 4 degrees centigrade. Does anyone know if there is still heat produced during the polymerization?
This seems to be in the realm of polymer chemistry and materials science. I'd appreciate any and all help, Thanks.
Try "M & R Optical" -- Sales, Service and Calibration of all Makes and Models. The persons name is Paul Musci. He services all of our optical microscopes (all of which are older pieces of equipment).
Here is the pertinent information: Paul Musci 64 Osgood Rd. P.O. Box 515 Charlton City ,MA 01508
Tel. (508)248-5684 Fax.(508)248-4658
OR PERHAPS you could try--} A place called PYE Metallurgical Consulting, INC. One of the things this business specializes in is used and refurbished equipment. Perhaps (maybe) they could get this objective for you, or at least point you in the right direction......
This sounds reminiscent of a beta backscatter detector we used to have. Except I think we only used it for coating thickness measurements (I never used it personally). You might want to check out ASTM B-567 Standard Test Method for Measurement of Coating Thickness by the Beta Backscatter Method.
Diane Ciaburri General Dynamics Pittsfield MA
} Tony Garratt-Reed {tonygr-at-mit.edu} } 10/09/01 10:59 AM } } } Hi, Listers! } } Many years ago, when (or even before) I was an undergraduate (that would } place it in the early '60s), my aunt managed the analytical lab at a } company that produced non-ferrous metals (principally, I think, bronzes } and brasses). As one does in such situations, she invited me to tour her labs } one day. } } One of the instruments I was shown was called a "Beta Probe". A piece of } the metal was ground and polished, and was then placed face down on an } aperture in the top plate, with what I now realize was an O-ring seal. A } few buttons were pressed, and after a little while some of the electronic } counters of the day (neon tubes with 10 electrodes) began spinning round. } My memory is that the operator had some range of ratios between the } recorded numbers within which he (it was, of course, a "he" in those days, } even though my aunt was the manager) could pass the sample as meeting } specifications. } } I surmise that this was an electron milli-probe (if you'll forgive my } coining of the term!) with preset spectrometers tuned to the elements of } interest in the specification. I would very much like to find out more } about this device. Happily, my aunt is still available to be asked, but } she knows nothing of the operation of the instrument. The people who ran } it have either died, or lost touch (the company failed twenty years ago). } } If anyone can give me more information about this instrument, I would be } very interested, and most grateful. } } Thanks, } } Tony } } } * * * * * * * * * * * * * * * * * * * * * * * * * * } * Anthony J. Garratt-Reed M.A., D.Phil. } * MIT, Room 13-1027 } * 77 Massachusetts Avenue } * Cambridge, MA 02139-4307 } * USA } * Phone: (617) 253-4622 } * Fax: (617) 258-6478 } *
{ { What about tetraethyl lead? Check into the catalysts (e.g. stannous octoate) used to polymerize the plga or other polymers. } }
I made a similar suggestion about 30 years ago, and was come down upon like a ton of bricks. Tetraethyl lead is not only extremely poisonous, it is rapidly absorbed by the skin. I understand that in companies where it is used, they have a paraffin shower, and the person is thrown under that clothes and all, no time to muck about.
Because of this, I looked up the work of J.Smid at Syracuse, NY, who uses bismuth compounds for the same purpose. From this work I learned about triphenyl bismuth, which appears to be a remarkably innocuous compound (remember the old days, when people used "bismuth" as an indigestion remedy?)
One of his published papers is:
Rawls HR, Granier RJ, Smid J, et al. Thermomechanical investigation of poly(methylmethacrylate) containing an organobismuth radiopacifying additive J BIOMED MATER RES 31 (3): 339-343 JUL 1996
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
} Can anyone offer any ideas on the best way to proceed with removing the } effects of charging on an image? I am referring to the streak across the } image manifested as a change in density. As an example, have a look at } this uncoated mite: } ...
Know that you can mask this image with a "gray" and set transparency such that the image will remain unchanged. I see several places in the original image where the horizontal banding can be sampled as a row of vertical pixels which range in brightness and is representative of the charging effect. Now apply this range of grays to the gray mask. If the sampling of banded grays includes 'white', then begin with a white mask and paint the banding across the mask and apply the mask with varying transparency. Not having tried this, it is possible the opposite effect might happen ... if so, then invert the mask.
Hi, Does anyone know if there was an accessory for the WILD M5 stereo microscope that provided a (small) range of continuously variable magnification. The feature is needed to get an exact overlay of video camera images from two different microscopes.
Thanks,
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
I'd love to pick up some used copies of SEM-related texts , such as "Electron Microscopy" by Bozzola and Russell and "Scanning Electron Microscopy and X-Ray Microanalysis" by Goldstein, Newberg, Joy), or other similar textbooks. If anyone has spare copies, please contact me in private e-mail.
Does any one use any other service companies for scope service of their Philips EM400T TEM?
Other than FEI.
Joanne Crudele
-----Original Message----- } From: McFaddin, Wade [SMTP:Wade.McFaddin-at-nextekinc.com] Sent: Tuesday, October 09, 2001 7:59 AM To: Microscopy-at-sparc5.microscopy.com
Nextek Inc. is seeking a qualified individual to join our analytical laboratory team as a Materials/Failure Analysis Engineer. The ideal candidate will have a B.S. in Materials Science, Chemistry, Engineering or equivalent, and a minimum 3 years experience in the micro-electronic field; or 5 or more years hands-on lab experience in the micro-electronic materials/failure analysis field. Knowledge in cross-sectional and destructive analysis of micro-electronic assemblies and devices, and experience with analytical instrumentation such as; SEM/EDS, Micro-FTIR, Thermal Analysis, Acoustic and Optical Microscopy is a plus.
To learn more about Nextek Inc. visit our web site at http://www.nextekinc.com/. For more information about this position, call or email the following:
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We are going to purchase a CCD camera for our CM20 STEM. I need comment on the AltraScan 1000 made by Gatan and KeenView DUAL DOCU IMAGING SYSTEM made by SIS. Any opinion is welcome.
We have a technician job, mostly related to repair and maintenance of our scopes and other equipment. Salary is $20k to $30k per year. It will be posted next week.
Applications should be done directly to the Univ. See their job website for the posting: http://www1.umn.edu/ohr/employ.html
The basic info is as follows:
Assist in an analytical laboratory consisting of electron microscopes, Ion Beam Accelerator/spectrometer, X-ray diffraction equipment, scanning probe microscopes, optical microscopes and other instrumentation.
0 50% Daily maintenance of above mentioned instruments and associated sample preparation equipment
0 35% Troubleshooting and repair of instruments, vacuum equipment, electronics and computers
0 15% Keeping labs clean, organized and adequately supplied
******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
Hello, I've been trying to carbon coat some samples for SEM in a denton vacuum evaporator. I used C-rods, one thinned from the original 4mm diameter to about 1mm for a short distance (1cm) touching a flat C-rod. I manage a few times to get 12-15 nm, but often the rod breaks, or shorts out before I can get even 5 nm deposited. I like using the high vacuum on the evaporator as you get a much better coating than from our sputter coater.
Can anyone send me some pointers as to how I can get a more controlled coating with one run of the evaporator? I have to almost go to maximum power to start getting deposition registered on the monitor. I tried changing the spring so it has a more even gentler push on the sharpened rod.
It's kind of strange that you have a problem. Your setup looks like OK. I would recommend the following:
-try 5 mm instead 10 mm for 1 mm dia part. -play with spring pressure and check that spring has enough "power" when the rods will short at the end of evaporation (very usual problem). -increase current slowly with a few stops. I would suggest 10A/10-20 sec is OK. -you may try graphite or carbon. Carbon is stronger mechanically and evaporated slower and with higher current than graphite. -both rods should be CO-AXIAL. -1 mm "tip" should be sharpened to the conical shape approx 90 deg.
Personally, I prefer 6 mm carbon rods. One of them - with 1 x5 mm tip on the end:
------------------I /-------------- I {== 6 mm dia ------------------I \-------------- -1x5mm
I wish you luck. Sergey
At 03:30 PM 10/12/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
A plastic part is steam sterilized and can have a residual water drop in a small depression in the part. All the water has to be removed before the mfg process can continue. The proposed solution is to apply pressurized air (or some other gas) to remove the water drop. However, when a jet of pressurized air is applied it will blow out most of the water but leave behind small 20-50 micrometer microdroplets. These are still a problem.
I thought that I could use a dye in the test droplet of water applied to the part before it is subjected to the proposed air blast in an experimental test setup. The idea would be that if any microdroplets remained I would be able to see them under a simple macroscope. However, for such things as food dye, when it dries, the color remains and I can't tell if it is wet or dry.
One of the acceptable situation may be that most of the droplet is blown off and the remaining microdroplets dry within seconds using heat lamp, dry air, low RH in the test area, etc. So what I'm looking for is a dye that is one color when wet and either another color when dry or no color at all (clear, or white powder). My first attempts have been with cobalt chloride but driving off all the mositure is difficult. Another possibility is using a fluorescent dye that fluoresces one color when wet and has a very much reduced fluorescence when dry or better yet a different color.
I certainly appreciate you help and all suggestions will be considered, even if you have a totally different idea on how to determine when all the water is gone.
A couple of additional points: Some evaporator give a choice of voltages and this is a important feature. For metal evaporation 10V gives better control then a higher voltage, but this is not suitable for carbon rods. Graphite and even more so carbon rods, are poorer conductors than metal wire or foils, consequently evaporation is much more likely to fail at the lower voltages. Check your evaporator and if you are using something like 10 volts, then see if you can tap the transformer at a higher voltage setting, I recommend 25 or 30V. Obviously the amps required for evaporation will change. If for daily operations changing the voltage is troublesome, then I would rather put up with a touchy control for metal evaporation then with carbon evaporation failures, i.e. rather use 25V for all evaporations then 10V. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, October 13, 2001 1:50 PM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Gordon, hello } } It's kind of strange that you have a problem. Your setup looks like OK. I } would recommend the following: } } -try 5 mm instead 10 mm for 1 mm dia part. } -play with spring pressure and check that spring has enough "power" when } the rods will short at the end of evaporation (very usual problem). } -increase current slowly with a few stops. I would suggest 10A/10-20 sec } is OK. } -you may try graphite or carbon. Carbon is stronger mechanically and } evaporated slower and with higher current than graphite. } -both rods should be CO-AXIAL. } -1 mm "tip" should be sharpened to the conical shape approx 90 deg. } } Personally, I prefer 6 mm carbon rods. One of them - with 1 x5 mm tip on } the end: } } ------------------I /-------------- } I {== 6 mm dia } ------------------I \-------------- } -1x5mm } } I wish you luck. Sergey } } } At 03:30 PM 10/12/01 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello, } } I've been trying to carbon coat some samples for SEM in a denton vacuum } } evaporator. I used C-rods, one thinned from the original 4mm diameter to } } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a } } few times to get 12-15 nm, but often the rod breaks, or shorts out before } } I can get even 5 nm deposited. I like using the high vacuum on the } } evaporator as you get a much better coating than from our sputter coater. } } } } Can anyone send me some pointers as to how I can get a more controlled } } coating with one run of the evaporator? I have to almost go to maximum } } power to start getting deposition registered on the monitor. I tried } } changing the spring so it has a more even gentler push on the sharpened } } rod. } } } } The rods I use after sharpening look like this: } } } } --------| |--------------- } } | | } } ---------| } } ---------| } } | | } } --------| |-------------- } } } } Hopefully the ascii art comes out in email. } } } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } } \ } } Gordon Ante Vrdoljak Electron Microscope } } Lab } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } phone (510) 642-2085 Berkeley CA 94720-3330 } } fax (510) 643-6207 cell (510) 290-6793 } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } }
I use the same method, and I have the same problem if I thin the rods down too much. Try turning it down to taper down to 2mm over 6mm of length, and make sure that the flat end of the other electrode doesn't have any debris left on it from previous use. For the flat-ended rod, I use 6.5mm diameter (I don't know whether it is graphite or carbon, I don't think it matters for this one).
In contrast to Sergey's experience, I have found that graphite needs to get much hotter to evaporate than does carbon, and the holder assembly all became red hot when I tried graphite.
Jim Darley at Proscitech sells both types.
My only relationship with him is as a satisfied customer.
good luck
rtch
} } Hello, } I've been trying to carbon coat some samples for SEM in a denton } vacuum evaporator. I used C-rods, one thinned from the original 4mm } diameter to about 1mm for a short distance (1cm) touching a flat } C-rod. I manage a few times to get 12-15 nm, but often the rod } breaks, or shorts out before I can get even 5 nm deposited. I like } using the high vacuum on the evaporator as you get a much better } coating than from our sputter coater. } } Can anyone send me some pointers as to how I can get a more } controlled coating with one run of the evaporator? I have to almost } go to maximum power to start getting deposition registered on the } monitor. I tried changing the spring so it has a more even gentler } push on the sharpened rod. }
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
We would like to upgrade our PC containing SemAfore 3.02 hooked with JEOL JSM5300. The old card is using the ISA however all the new boards do not have the ISA but PCI only. So we've asked JEOL people to get ISA extender. After getting the new 1 Ghz PC, hooking the ISA extender, hooking the card, got everything hooked up ... I think the engineer told me ... "very-very-very low signal". So we looked around and found one motherboard with one ISA and the rest PCI. Hooked everything up. Still the same thing happened. The engineer told us that we need to purchase the USB type which costs a hefty sum in order to use with the new PC. So now we are back to the old PC. My worry is what happen when the old PC konk.
Any suggestion or somebody experienced this? btw, the engineer also tried with SemAfore 4.0 and it did not work. } } Regards, } } Fauzi } Malaysian Rubber Board }
_________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.com address at http://mail.yahoo.com
Gordon, The most obvious thing, that no one has mentioned yet, is that the non-sharpened rod should be shaped at a 45 degree angle with the face towards your sample. The one mm rod should hit roughly in the center of the 45 degree face. This allows a better distribution of carbon towards your sample. 1 cm of 1mm rod seems awfully long to me. The trade-off is distance from sample vs heating of sample. Closer gives you more rapid coating due to the larger solid angle. Too close gives very intense heating. Too far away can also impart too much heat because it can take so long to get the thickness you're looking for. My understanding is that 9-10 cm should be a good distance. Someone with more math skill than I can probably tell you how many mm of 1mm diameter carbon you need at that distance to get 5, 10 or 15 nm coating.
Ken Converse owner Quality Images third party SEM service Delta, PA
Gordon Vrololjak wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I've been trying to carbon coat some samples for SEM in a denton vacuum } evaporator. I used C-rods, one thinned from the original 4mm diameter to } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a } few times to get 12-15 nm, but often the rod breaks, or shorts out before } I can get even 5 nm deposited. I like using the high vacuum on the } evaporator as you get a much better coating than from our sputter coater. } } Can anyone send me some pointers as to how I can get a more controlled } coating with one run of the evaporator? I have to almost go to maximum } power to start getting deposition registered on the monitor. I tried } changing the spring so it has a more even gentler push on the sharpened } rod. } } The rods I use after sharpening look like this: } } --------| |--------------- } | | } ---------| } ---------| } | | } --------| |-------------- } } Hopefully the ascii art comes out in email. } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } } }
Two postdoctoral positions (see below) are available immediately. If interested, send a CV including the names of referees to ldm3-at-apollo.numis.nwu.edu
1. Quantitative Electron Microscopy
A postdoctoral position is available in the area of quantitative TEM/TED of materials starting immediately. The research project focuses on developing quantitative methods of solving structures from diffraction patterns and HREM images. Analysis of diffraction patterns will be via Direct Methods to recover estimates of the phases; analysis of HREM images will be to develop methods to routinely perform translational/rotational averaging. The work will involve collaboration with scientists at LBL and elsewhere. A good background in computer programming in fortran or C is required, as well as an understanding of dynamical diffraction theory. Prior experience in HREM or Direct Methods would be an advantage.
2. Surface Microscopy
The position would involve work using the unique HREM/Surface Science facility at Northwestern University (see http://www.numis.nwu.edu). The primary research area will be metalic nanostructures on semiconductor surfaces, although elements of the work will also overlap with other ongoing projects involving magnetron deposition of quasicrystalline thin films, hard coatings and oxide surface structures. A strong background in basic TEM techniques as well as some familiarity with thin film growth, surface science and other characterization techniques such as XPS are important.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
I have used approx. 20 different Denton 502's for evaporation (and FE) over the last 30 years. I also recondition and sell them. Since you are maybe using one of them "carbon fiber Flash Evaporators"? sputter coaters, the speed to evaporate in HV has to be SLOW. Have sufficient spring pressure, s l o w l y bring up the current, watch the rod (with welder's glasses or cover the bright zone by viewing over the holder bar, DONT LOOK INTO THE BRIGHT SPOT!! Of course you know that!) until some fine sparks appear, immediately reduce the current below sparking and watch your indicator for appropriate coating. It works 99.9% of the times. Regards, Markus F. Meyenhofer Microscopy Labs
Colleagues, can somebody recommend (by own positive experience!) an antibody against collagen, e.g. Type IV-Collagen ? Antibody should work in Immuno-EM, preferably tolerating glutaraldehyde fixed material.
Thanks for direct short, informal e-mail to me.
Peter
************************************************** please reply to: peter.heimann-at-biologie.uni-bielefeld.de ("E-MAIL")
Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : xx49(0)521-106-5654 WEB-Site: http://www.uni-bielefeld.de/biologie/Entwicklungsbiologie WEB-Site: http://www.uni-bielefeld.de/SFB549 ******************************************************
Dear Gordon, I have a very old JEOL evaporator and what I found was: 1. I sharpen my 5 mm dia. rods to 1 mm for no more than 4 to 5 mm. Flatten the rod it rests on with the edge of a glass slide. 2. Use pressed carbon, not graphite. It heats more readily by resistance. 3. Carefully align and tighten the rods before you let the spring-loaded one rest against the center of the fixed one. This avoids putting a twist in your thinned, weak rod. 4. Don't knock the rods as you are putting the lid on. I find you have to turn up the current slowly, watching through welding goggles, just until the tip starts to spit sparks. Leave it there and it will climb up by itself after that. At 03:30 PM 10/12/01 -0700, you wrote: } Hello, } I've been trying to carbon coat some samples for SEM in a denton vacuum } evaporator. I used C-rods, one thinned from the original 4mm diameter to } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a } few times to get 12-15 nm, but often the rod breaks, or shorts out before } I can get even 5 nm deposited. I like using the high vacuum on the } evaporator as you get a much better coating than from our sputter coater. } } Can anyone send me some pointers as to how I can get a more controlled } coating with one run of the evaporator? I have to almost go to maximum } power to start getting deposition registered on the monitor. I tried } changing the spring so it has a more even gentler push on the sharpened } rod. } } The rods I use after sharpening look like this: } } --------| |--------------- } | | } ---------| } ---------| } | | } --------| |-------------- } } Hopefully the ascii art comes out in email. } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab I hope this helps. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
The DTSA manual describes the method of "smoothing" a spectrum as a 5, 7, or 9 point Savitsky-Golay polynomial. Does anyone know how this is applied. Is it similar to passing a filter through the data with weighted coefficients? I would appreciate if anyone can give me the algorithm. Thanks in advance.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
In a message dated 10/15/01 2:27:04 PM, walck-at-ppg.com writes:
} The DTSA manual describes the method of "smoothing" a spectrum as a 5, } 7, or 9 point Savitsky-Golay polynomial. Does anyone know how this is } applied. Is it similar to passing a filter through the data with weighted } coefficients? I would appreciate if anyone can give me the algorithm. } Thanks in advance
That is exactly what it is. The S&G coefficients allow fitting polynomials of various degrees (e.g. a quadratic polynomial) to data in order to smooth it. The 7 point coefficients for a quadratic, for example, are -.0952 .1429 .2857 .3333 .2857 .1429 -.0952 to be multiplied by each group of the histogram values in order to produce one new point at the center of the 7. This is then repeated at every point to generate a new, smoother curve.
I would like to announce the debut of the Food Structure and Functionality Discussion group at the AOCS (Americal Oil Chemists Society) website, webaddress:
http://www.aocs.org/ubbcgi/ultimatebb.cgi
This discussion group is open to all agri-food researchers from Government, Industry and universities to discuss topics of interest, problems, sample preparation, etc. much like the function of this Microscopy listserver, as well as being a source of information for the activities of the Food Structure and Functionality Forum - a division of AOCS, and its yearly Symposium.
We want to make it an interesting and valuable resource for anyone who visits it. Please visit the site today and start or join a discussion! The site is up and ready for you to use.
Kind regards,
Paula Allan-Wojtas, Chair Food Structure and Functionality Forum - a division of AOCS
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
I was going to take a look at your book and see what they were across the center row for an image Gaussian smoothing filter if nobody replied. Somebody is currently borrowing my copy however. (Fred, if you are reading this, it is time to return it.)
Let me bother you with another question and I made it public on the Listserver. How do you handle the ends of the data array, i.e. in a 1024 array, the indices 1-3 and 1022-1024. When you apply the coefficients, they sum to one, so you can't just use some of them.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Monday, October 15, 2001 2:40 PM To: walck-at-ppg.com; microscopy-at-sparc5.microscopy.com
In a message dated 10/15/01 3:49:53 PM, walck-at-ppg.com writes:
} I was going to take a look at your book and see what they were across the center row for an image Gaussian } smoothing filter if nobody replied. Somebody is currently borrowing my copy however. (Fred, if you are } reading this, it is time to return it.)
Yes, they are in the book. Tell him to buy his own copy!
} Let me bother you with another question and I made it public on the Listserver. } How do you handle the ends of the data array, i.e. in a 1024 array, the } indices 1-3 and 1022-1024. When you apply the coefficients, they sum to } one, so you can't just use some of them.
Edges are always a problem. The simplest solution is to consider them a mirror, so that the -1, -2, -3 positions have the same values as +1, +2, +3 (and the same thing at the other end). The other common choices are to perform some kind of extrapolation on the data at each end (the simplest of which is linear) or to make your addressing wrap around (i.e., -1 = 1023, -2 = 1022, etc.) which is not appropriate for most cases.
In any case, you can expect to lose some data at the ends (or for images, the edges).
Fellow microscopists I have a two dye sublimation printers that are gathering dust in my lab. If anyone is interested, you can make me an offer. The the first printer circa 1994 Seiko ColorPoint professional 300 dpi dye sublimation unit with an interface box to allow screen captures from a computer monitor. The unit includes a spare ribbon and 50 sheets of paper. The second is a Fargo primera pro elite 300 dpi resolution. This unit is a win printer, and will only work with Windows 95 or 98 (which I no longer use). The reason I'm not using them is that they simply cost too much to run. I replaced both of them with a networked inkjet printer that is much cheaper to run and suits my requirements just as well.
Both include manuals and are for sale as is. If anyone is interested they can drop me a note. -- Glenn
=============================================================================== Glenn Poirier 3450 University St, rm. 238 MicroAnalytical Laboratory Montreal, Qc Earth and Planetary Sciences tel (514) 398 6774 McGill University fax (514) 398 4680 email: glennp-at-eps.mcgill.ca http://castaing.eps.mcgill.ca
++ Millenium hand and shrimp ++ ===============================================================================
A physicist colleague asked if I knew how to make microscope slides repellent so that dust and the paper fibres he is studying would not adhere to them. Apart from applying a thin coat of some non-polar material, I didn't have any useful suggestions - I'm usually trying to make things stick, not vice versa.
Anyone have a more useful and/or more specific suggestion or protocol?
Thanks, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
It depends what you want: if you need to prevent ionic interactions in the solution (like ion-pairs) make glass hydrophobic by siliconization. If you want to remove static electricity (which is usually attracts the dust) you may have to make glass electro-conductive (make it wet in simply case, poly-lysine may work as well). Both recommendations are quite controversial because siliconization make glass hydrophobic and it increase the chance of electization when dry. I would suggest that there is no solution for your case: dust is heterogeneous and some part of this nasty thing will find the way to contaminate your sample. In this battle the dust is always a winner.
Paper fibers, cellulose, has a lot of -OHs, so the nature of adhesion is polar/ion (in the water solution). Siliconization would help.
Sergey
At 09:26 AM 10/16/01 +1000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
on 10/16/01 7:26 AM, Rosemary White at Rosemary.White-at-pi.csiro.au wrote:
} } A physicist colleague asked if I knew how to make microscope slides } repellent so that dust and the paper fibres he is studying would not adhere } to them. Apart from applying a thin coat of some non-polar material, I } didn't have any useful suggestions - I'm usually trying to make things } stick, not vice versa. } } Anyone have a more useful and/or more specific suggestion or protocol? } Dear Rosemary, I don't know whether this will work for the particular application, but many scientific supply houses make a silicone solution designed to make glass hydrophobic. I think Sigma's is called Siliclad. Good luck to a fellow physicist. Yours, Bill Tivol
Same situation here - we usually want things to stick - but poly-lysine is out. What charge do the particles have - maybe there is something with the same charge that would repel?
My other thought is PTFE/Teflon in it various presentations ("thin coat of some non-polar material"). I use it on the moving parts of my bicycle as a solution in spray form and also applied as a liquid solution. I can't remember what the solvent/solvent mixture is but I would guess that it is alcohol based. I have never tried it on microscope slides but is might work.
Would it do the job?
Med vänliga hälsningar/With best regards
Gareth
Då du älskar, älskar med glödande hetta. Då du hatar, hata i flammande blixtar. Då du festar, festa som om vardagen inte fanns. Då du går, gå fort, men lämna alltid dörren på glänt......
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
Hi I would like to thank everybody very much for your advice on the subject of { fluorescence microscopy and TEM on same sample} . I am working on the procedure and may come back with further questions.
Glass microscope slides can be made hydrophobic using hexamethyldisilAzane (HMDS) (Aldrich Chemical 37,921-2). Simply place the separated slides in a container like a wide mouth glass jar with an aluminum or Teflon lined cap. Add a few drops of HMDS and it will vaporize overnight in the sealed jar and react with the silanols on the surface. I prefer to heat the jar in an oven at 70° overnight to drive off residual ammonia. Leave the jar slightly cracked open. The next day, in a hood, open and remove the slides. They will now bead water. This will be a molecular level coating and invisible.
Things like Rainex® and Aquapels® will work but can't be applied in the vapor state. You do not want to create artifacts (streaks) on the slides, so use the vapor state application of hexamethyldiSILAZANE (HMDS). HexamethylsilOXane is NOT the same material. HMDS is simple, easy to apply, and safely handled.
Use gloves, a hood, and respirator when handling these materials. Let the dropper and the opened jar with HMDS evaporate overnight in the hood and then place them in the trash the next day. Do not breath HMDS vapors at all! Read the MSDS information on handling.
I had a 'never opened bottle and wax sealed bottle' of chlorosilane spontaneously explode in a closed cabinet. It formed lots of white smoke and HCl. Nobody was around at the time it happened. This will make you a believer in using HMDS when possible.
These are my opinions only but based on my experiences.
Paul Beauregard Senior Research Associate Monroeville, PA 15601
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-pi.csiro.au] Sent: Monday, October 15, 2001 7:26 PM To: Microscopy-at-sparc5.microscopy.com
Dear all,
A physicist colleague asked if I knew how to make microscope slides repellent so that dust and the paper fibres he is studying would not adhere to them. Apart from applying a thin coat of some non-polar material, I didn't have any useful suggestions - I'm usually trying to make things stick, not vice versa.
Anyone have a more useful and/or more specific suggestion or protocol?
Thanks, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Wow! is this ancient history! My first paper ever was about the determination of the thickness of carbon films correlated with optical density. Which meant that I did a _lot_ of films, with different length carbon rods (using an old Kinney evaporator actually). The primary result was that OD and thickness have a linear relationship from about 5nm to 100nm. (And right now I am having a senior moment and not remembering the exact correlation--and where is that paper now that I need it???) On the practical side, 1mm carbon rod would give a 10nm film, and as I remember the correlation was fairly linear between 0.5mm (5nm) up to 5mm (50nm). I simply use the old "folded filter paper" method for evaluation, knowing that an OD of about 1 gives a very heavy film--probably about 100nm. As Ken says, geometry has a lot to do with the final thickness deposited, but once you work it out for your system it should be fairly consistent. If my brain functions and I can get some of these tired neurons to fire (for more precise information), I'll forward it along. Roger
On Mon, 15 Oct 2001 08:03:44 -0400, Ken Converse wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Gordon, | The most obvious thing, that no one has mentioned yet, is that the | non-sharpened rod should be shaped at a 45 degree angle with the face | towards your sample. The one mm rod should hit roughly in the center | of the 45 degree face. This allows a better distribution of carbon | towards your sample. 1 cm of 1mm rod seems awfully long to me. The | trade-off is distance from sample vs heating of sample. Closer gives | you more rapid coating due to the larger solid angle. Too close gives | very intense heating. Too far away can also impart too much heat | because it can take so long to get the thickness you're looking for. My | understanding is that 9-10 cm should be a good distance. Someone with | more math skill than I can probably tell you how many mm of 1mm diameter | carbon you need at that distance to get 5, 10 or 15 nm coating. | | Ken Converse | owner | Quality Images | third party SEM service | Delta, PA | | Gordon Vrololjak wrote: | | } ------------------------------------------------------------------------ | } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
| } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | } -----------------------------------------------------------------------. | } | } | } Hello, | } I've been trying to carbon coat some samples for SEM in a denton vacuum | } evaporator. I used C-rods, one thinned from the original 4mm diameter to | } about 1mm for a short distance (1cm) touching a flat C-rod. I manage a | } few times to get 12-15 nm, but often the rod breaks, or shorts out before | } I can get even 5 nm deposited. I like using the high vacuum on the | } evaporator as you get a much better coating than from our sputter coater. | } | } Can anyone send me some pointers as to how I can get a more controlled | } coating with one run of the evaporator? I have to almost go to maximum | } power to start getting deposition registered on the monitor. I tried | } changing the spring so it has a more even gentler push on the sharpened | } rod. | } | } The rods I use after sharpening look like this: | } | } --------| |--------------- | } | | | } ---------| | } ---------| | } | | | } --------| |-------------- | } | } Hopefully the ascii art comes out in email. | } | } | } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ | } Gordon Ante Vrdoljak Electron Microscope Lab | } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall | } gvrdolja-at-nature.berkeley.edu UC Berkeley | } phone (510) 642-2085 Berkeley CA 94720-3330 | } fax (510) 643-6207 cell (510) 290-6793 | } | } | } | } | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
A local research physician's widow has contacted me to find out whether her husband's microscope has any value or is still useful. It's a Nikon model L-Ke (possibly with a camera attachment). Can anyone help with information?
Many thanks, Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Just wanted to confirm our schedule and request driving instructions/hotel info for the Hitachi site. Also wondering which airport is best to fly into (?).
I need to produce clear cut high resolution pictures of the peptidoglycan layer of E. coli. I am looking for a staining method that shows some specificity for peptidoglycan (may be some method for polysaccharides, I guess) while allowing high resolution immages.
Preferably the method should work with Epon-Araldite embedded material fixed in glutaraldehyde and osmium.
Does anyone have suggestions for such a reliable stainning method?.
Dr. A.P. Alves de Matos Curry Cabral Hospital Lisbon apmatos-at-ip.pt
We were wondering if anyone else has run into this problem: we bought a Hitachi VP S-3000N SEM. They offer a 125 mm multiple specimen holder - essentially an aluminum disc on a small column that has 33 holes in it to hold 33 stubs. We bought it and have now discovered that none of the commercial vendors sell pin-type stubs that will actually fit into these holes! The majority of stubs have pins that are 3mm in diameter. The holes are only 2 mm in diameter. EMS can make the stubs at 13 cents a piece but the minimum order is 100,000. Has anyone out there found a source of stubs at a reasonable price (Hitachi offered to sell them at approximately $25.00 a piece!) or are people just drilling the holes in the holder to make them bigger? I'd really like to hear how other people are handling this!
Lesley
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
I made my own multi stub holder. But if I had what you have I'd head down to the shop and drill/ream out those holes or have a competent shop guy do it. Maybe 20-30 minutes of work, tops.
} We were wondering if anyone else has run into this problem: we bought a } Hitachi VP S-3000N SEM. They offer a 125 mm multiple specimen holder - } essentially an aluminum disc on a small column that has 33 holes in it to } hold 33 stubs. We bought it and have now discovered that none of the } commercial vendors sell pin-type stubs that will actually fit into these } holes! The majority of stubs have pins that are 3mm in diameter. The } holes are only 2 mm in diameter. EMS can make the stubs at 13 cents a } piece but the minimum order is 100,000. Has anyone out there found a } source of stubs at a reasonable price (Hitachi offered to sell them at } approximately $25.00 a piece!) or are people just drilling the holes in } the holder to make them bigger? I'd really like to hear how other people } are handling this! } }
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Applications for an NEI-supported post-doctoral position to study M cells in the mammalian conjunctiva using LM & EM techniques are being accepted.
M cells are classically found overlying the mucosal lymphoid follicles (Peyer's Patches) of the intestinal tract. The importance of M cells in the mucosal immune response has only recently been appreciated. M cells have a distinctive morphology that includes an invagination of their basolateral membrane to form a "pocket" filled with macrophages, lymphocytes and other antigen presenting cells. The apical portion of the M cell forms thin bridges that separate the extracellular world from the underlying lymphoid cells. This narrow bridge can result in the distance from the apical membrane to the underlying pocket of less than 2 microns. The M cell acts as an antigen sampling cell by capturing souble and particulate lumenal antigens, transcytosing them across the thin cytoplasmic bridge) and releasing them into the subcellular pocket filled with macrophages and other antigen presenting cells. Endocytosis of antigens by M cells is thought to be the first step in generating a mucosal immune response. There is considerable interest in the development of immunogens targeted to M cells to develop immunity against diseases such as cholera. It has also been realized that opportunistic pathogens (e.g., HIV, Salmonella, Shigella) selectively bind to intestinal M cells and exploit the transcytosis process to allow them to penetrate the epithelial barrier.
M cells have also been found to occur in tonsils, bronchi, and the nasal cavities. The presence of M cells in the ocular conjunctiva is much more controversial. In collaboration with Dr. Cecil Moore, DVM (a veterinary ophthalmologist and chair of Veterinary Medicine & Surgery, MU Veterinary School), my lab has recently found LM, TEM, and SEM evidence of cells with the highly distinctive morphology of M cells in the conjunctiva of dogs (paper under review). Another group found similar views in the Guinea pig. The bulk of the literature claims M cells don't exist in the eye. This is mostly because no one has ever tested the ability of follicular associated epithelium in the eye to selectively transport antigens. Our new grant is designed to test the ability of these putative M cells to selectively bind and transport antigens of pathological importance. We intend to screen the conjunctiva of multiple mammalian species to prove the widespread existence of conjunctival M cells. We will also be testing whether ocular immunization is superior to immunization at other sites in regards to generating an ocular immune response. Candidates should have experience in light and/or electron microscopy.
The project is mostly LM, TEM and SEM with some immunology later on. Funding for the postdoc is $28,260 plus full benefits. There is funding for 3 years but departmental policy requires offers to be made for 1 year with re-appointment for subsequent years dependent on satisfactory progress. The postdoc is available immediately but I would be willing to wait a reasonable amount of time for the right candidate. Send CV and names of three references by e-mail to PhillipsT-at-missouri.edu. -- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I am seeking feedback--good and bad--about a planned x-ray detector system purchase.
I'm doing semiconductor work and need light element detection (down to B). I do not have LN2. As a result, I've narrowed the candidates to Noran's cryo cooled 10mm detector and their Quest software system. The detector would be mounted on an Amray 1910 FESEM.
The Noran system comes with its own PC. I'd rather have it integrated into my SEM control PC. But I suppose this would work out OK in the long run. Noran uses a Compaq PC with dual Ethernet ports. One port talks to the pulse counter and detector interface. I may try to use the other Enet port to connect the SEM PC.
I would appreciate any feedback from current users of this product. I'm wondering about reliability and up-time. Also, how are these systems warranted and maintained? Off-list responses are welcomed.
Dear Listers I am presently undetaking an Honours degree at La Trobe University, Victoria, Australia.I have been researching flower pollen using a Scanning Electron and Confocal microscopes. I am particularly interested to hear your views on the following question: Do you see a link between science and art? If so, how and why? Thank you for considering my question. Yours sincerely Judi Bowden La Trobe University Victoria Australia
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (j.miyan-at-umist.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, October 16, 2001 at 12:07:24 ---------------------------------------------------------------------------
Email: j.miyan-at-umist.ac.uk Name: Jaleel Miyan
Organization: UMIST
Education: Graduate College
Location: Manchester, UK
Question: We are looking for a method to stain for proteoglycans in sections of rat brain. I had heard that you can use a critical eletrolyte method using alcian blue and varying magnesium concentrations.
Can you direct me to a protocol and/or comment on the accuracy of this and any other possible method.
This is a reply to all the people who have been asking me about what kind of success I have had with the cell wall dyes.
First of all, I am trying to stain 1 micron thick sections of LR White-embedded cotyledons of Vicia faba. I got really nice staining with a 1% solution (I know - it's pretty strong!) for 2 min (no heating). I washed the sections after staining in running water for 15 min to remove any unbound stain. The stain instantly got weaker when excited (green filter) if I mounted the sections in water or 50% glycerol. So I mounted them in Zeiss immersion oil, which gave me a really, really nice clear image. The only problem was that, as was pointed out by Susan Joa, the dye binds to 1-4 Beta Glucans, so the starch grains were fluorescing pretty strongly. However, in my opinion the image I got was much better than any calcofluor image I have ever taken. In addition, the nuceli were lightly stained (does anyone know why this is?).
Hi: We are looking (again) for a used Zeiss 10 TEM for the Los Angeles area. Please let me know if you have one for sale or if you know of somebody selling. Thank you, Peter Jordan 909 302-9130
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mryder-at-brookes.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, October 17, 2001 at 05:31:30 ---------------------------------------------------------------------------
Question: Why do my wholemount lowish power root photographs sometimes come out yellow, on the same film, with the same lighting. Sometimes the same root taken a few minutes later will be yellow when it was the usual white on a previous frame.
After my company bought a 3000N, I recognized that the Hitachi sample holder system was relatively expensive. So I designed a 50 mm holder with space for 8 12-13 mm dia. relatively inexpensive Zeiss stubs and had a local machine shop make 7 of them. That included set screws for the sample stubs and an appropriate center hole to screw in a Hitachi base. The 3000N memory system was used to remember the centerpoint of each of the 8 sample sites so moving from sample to sample is easy. The cost was easily recovered considering the high cost of the Hitachi mount system. All of the users were assigned a holder and it's worked without a hitch ever since.
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We were wondering if anyone else has run into this problem: we bought a Hitachi VP S-3000N SEM. They offer a 125 mm multiple specimen holder - essentially an aluminum disc on a small column that has 33 holes in it to hold 33 stubs. We bought it and have now discovered that none of the commercial vendors sell pin-type stubs that will actually fit into these holes! The majority of stubs have pins that are 3mm in diameter. The holes are only 2 mm in diameter. EMS can make the stubs at 13 cents a piece but the minimum order is 100,000. Has anyone out there found a source of stubs at a reasonable price (Hitachi offered to sell them at approximately $25.00 a piece!) or are people just drilling the holes in the holder to make them bigger? I'd really like to hear how other people are handling this!
Lesley
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
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Dear Judy,
While I'm not a microscopist, I think there is a strong link in that some of the work one does for science can certainly become art.
If you are interested in further exploring the connection, I suggest you contact David Scharf of Los Angeles CA. His email address is dscharf-at-pacbell.net. David is an Artist who produces color SEM photomicrographs and his work is outstanding. In fact, he recently won an EMMY award for his work on a National Geographic documentary film called "Body Snatchers".
Sincerely,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: "01151938-at-mrc.vic.edu.au"-at-sparc5.microscopy.com [mailto:"01151938-at-mrc.vic.edu.au"-at-sparc5.microscopy.com] Sent: Tuesday, October 16, 2001 7:10 PM To: Microscopy-at-sparc5.microscopy.com
Dear Listers I am presently undetaking an Honours degree at La Trobe University, Victoria, Australia.I have been researching flower pollen using a Scanning Electron and Confocal microscopes. I am particularly interested to hear your views on the following question: Do you see a link between science and art? If so, how and why? Thank you for considering my question. Yours sincerely Judi Bowden La Trobe University Victoria Australia
Is there a procedure for keeping a pellet of cells together (as an intact pellet) while processing it for thin sectioning? I am working with virus infected cells removed from the substrate via trypsin and desire to keep the pellet densely packed and intact for subsequent processing steps. The embedded pellet will then be thin sectioned. Thanks in advance.
Tom Doman Animal Diagnostic Laboratory Penn State University
I know they sell epi-fluorescence attachments for stereoscopes but I am wondering how good they are in a practical sense. I want to detect clusters of FITC-tagged bacteria clustered on large pieces of mucosa measuring approximately 2 x 4 cm. Does anyone out there have any experience in this type of work. TIA, tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} } } Email: j.miyan-at-umist.ac.uk } Name: Jaleel Miyan } } Organization: UMIST } } Education: Graduate College } } Location: Manchester, UK } } Question: We are looking for a method to stain for proteoglycans in } sections of rat brain. I had heard that you can use a critical } eletrolyte method using alcian blue and varying magnesium } concentrations. } } Can you direct me to a protocol and/or comment on the accuracy of } this and any other possible method. } } MAny thanks } } Jaleel } } --------------------------------------------------------------------------- Jaleel There are a variety of cationic dyes, which because of their charge, will bind to proteoglycans, and that are electron dense. Many moons ago, I used alcian blue, ruthenium red, safranin O, and cetyl-pyridinium chloride (?) to contrast the ECM in heart muscle (see Robinson, et al, lab. Invest. 49(4):482-97 1983.) Also, in the 1970's and 1980 the Simionescu's had a lot of publications using various cationic dyes.
good luck, Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
You can resuspend your cell pellet in a few drops of 22 percent bovine albumin prior to glutaraldehyde fixation. Then, when you add the glut., the BSA proteins will crosslink and become a solid chunk which can then be processed as a piece of tissue.
An alternate method that I use is to resuspend the pellet and then centrifuge at each step of the prep. I routinely process virally infected cell monolayers this way. I like to embed in Spurrs rather than Epon because of its lower viscosity, allowing for easier pelleting. I do not use trypsin to lift the cells. Rather, I fix and post-fix the cells right in the culture dishes. Then, after the osmium rinses, I scrape the monolayers off of the dishes and pellet them in Eppendorf tubes for the remainder of the process.
I hope this helps!
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 FAX (734)763-1166 dsoren-at-umich.edu
On Wed, 17 Oct 2001, Tom Doman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleges, } } Is there a procedure for keeping a pellet of cells together (as } an intact pellet) while processing it for thin sectioning? I am working } with virus infected cells removed from the substrate via trypsin and } desire to keep the pellet densely packed and intact for subsequent } processing steps. The embedded pellet will then be thin sectioned. } Thanks in advance. } } Tom Doman } Animal Diagnostic Laboratory } Penn State University }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleges, } } Is there a procedure for keeping a pellet of cells together (as } an intact pellet) while processing it for thin sectioning? I am working } with virus infected cells removed from the substrate via trypsin and } desire to keep the pellet densely packed and intact for subsequent } processing steps. The embedded pellet will then be thin sectioned. } Thanks in advance. } } Tom Doman } Animal Diagnostic Laboratory } Penn State University
After fixing, pellet it down in the buffer in an eppendorf tube remove the buffer and add a drop of low melting point agarose made up in buffer (add no more than the same size of pellet) use a warmed pin (37 degrees) or long needle to swirl the pellet into the agarose before it sets. When set use a razor blade to cut off the end of the eppendorf to get at the pellet poke out the pellet onto a drop of buffer on dental wax or parafilm chop the agarose pellet into 1 mm squares put into a scintillation vial and process as normal for tissue pieces.
If the layer of cells is almost invisible, cover with a tiny amount of low melting point agarose set by cooling finish the processing in the eppendorf as long as the low melting point agarose does not lift off the bottom of the tube but traps the cells in place. In this case the thickness of the LMP agarose should allow penetration of the chemicals so should be very thin.
Elaine
-- Dr. Elaine Humphrey Director, Biosciences Electron Microscopy Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca website: www.emlab.ubc.ca
Remove the cells from the substrate (if you're dealing with an enveloped virus, you might want to use means other than trypsin).
Pellet cells gently (2000 x g for 10 min)
Resuspend in glut in microfuge tube.
Swirl for 30 sec, then pellet again.
Let pellet sit without resuspending to fix. Time will depend on pellet size.
After 1 to several hours, remove fix with pipet; cut end off plastic tube with razor blade. Holding tip with forceps under dissecting, scope scoop out pellet with flattened and pointed applicator stick.
Dry further with pie-shapped wedge of filter paper by touching only the tip to the cell mound. The consistency should be that of cooked oatmeal. If the pellet is not dry enough, cells will disperse in the agar and be hard to find in the EM. If it is too dry, ultrastructure will suffer.
Divide the pellet into clumps about 1 mm cubed with a razor blade or spatula and surround them with molten agar*.
Let harden, and wash in buffer. Proceed with osmium, etc., as though you had tissue blocks.
*Mix 1 g purified agar with 100 ml water (or 0.1 g in 10 ml); melt by boiling--watch out, it easily boils over. Put into small tubes; cap or cover with Parafilm. Store at 4 degrees C. When needed, melt in beaker of boiling water. Let cool till barely warm to touch--don't cook the cells. (Agar solidifies at about 36 degrees and melts at about 95 degrees.)
Do not put agar-encased cells into glut. It will cross-link the agar so that subsequent solutions don't penetrate. However the remaining glut in the (unwashed) cells prior to encasing in agar won't hurt.
Good luck.
Sara Miller
On Wed, 17 Oct 2001, Tom Doman wrote:
} Date: Wed, 17 Oct 2001 10:14:22 -0500 } From: Tom Doman {jtd1-at-psu.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Cell pellet processing for TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleges, } } Is there a procedure for keeping a pellet of cells together (as } an intact pellet) while processing it for thin sectioning? I am working } with virus infected cells removed from the substrate via trypsin and } desire to keep the pellet densely packed and intact for subsequent } processing steps. The embedded pellet will then be thin sectioned. } Thanks in advance. } } Tom Doman } Animal Diagnostic Laboratory } Penn State University } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts dropping. The problem developed over time. In the beginning, it would drop by very little and go back up again. Later on, it started dropping by a lot more and it took longer to get back up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing this problem? Any tips or suggestions? We appreciate the help and thank you all in advance.
I am a little rusty on this, but when I did my masters in interdisciplinary arts, I used the SEM. The connection I used between science and art was creativity. There are many fields that we can be creative in...science and art are two of them. We consider art the mother of creativity. Yet, we live in a scientific/technological paradigm these days more than a religious paradigm like the Middle Ages. Those main religious and scientific paradigms overlap(think of interlapping rings much like the Olympic rings) into other disciplines like medicine, art, history, and law. The thread that keeps it all woven together is creativity. There are many definitions to clarify, but I believe looking at science and art as part of a bigger construct fueled by creative energy will help you with the larger picture. The more immediate picture like what is science and what is art and what is creative remains for us to define and redefine. How we define it is by the tools we use, the message we seek, the role of transformation and discovery in what we produce. Good Luck!
We do this all the time with cell suspensions. Pellet the live cells (~1,000 rpm for 10 minutes), carefully decant the supernatant and add cold 3% glutaraldehyde, being careful not to disturb the pellet. After about an hour of fixation, you should be able to dislodge the pellet and process it as if it were a piece of tissue.
Hope this helps.
Jerry Gagne Senior Biologist Department of Microscopy and Microanalysis Abbott Laboratories (847) 938-5023
We have a student starting to work with human cerebral blood vessels obtained from cadavers.
A literature search has revealed little information on working with cadaver material at the TEM level.
The questions from this student are;
1. Does anyone know of any publications resulting from TEM investigations of samples from human cadavers, in particular cerebral blood vessels and brain / nervous tissue ?
2. The standard EM fixation texts talk about not using formalin to fix for TEM and emphasize speed of primary fixation. My initial results indicate that the ultrastructure of the cadaver cerebral blood vessels has remained unexpectedly intact.
Has anyone else experienced good ultrastructural results in the TEM with cadaver material, embalmed in 10% Neutral Buffered Formalin and after an initial post mortem delay of up to 24 hours ?
- What ultrastructural changes (at the organelle level) did you expect and then what actual ultrastructural changes did you see ?
3. What ultrastructural changes would you expect from such material if it has been in 10% Neutral Buffered Formalin for up to 6 months prior to processing and examination ?
On behalf of our student, thanks for your help.
Allan
-- ------------------------------------------------- Allan Mitchell Technical Manager Otago Electron Microscope Centre C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
----- Original Message ----- } From: {mryder-at-brookes.ac.uk} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, October 17, 2001 8:00 AM
Dear Listers,
We have recently got a free set of cryo-attachment (FC 4) for our old Reichert-Jung ultramicrotome. However, there are two parts missing. One is the alloy steel bridge, which connects the specimen to the ultramicrotome's specimen arm. The other is the steel guide rails, which are used for the mounting of the cryo-chamber. Anyone has these parts or whole cryo-attachment to give away or sell? Please let us know.
Thanks Cheng
----------------------------------------------------- Cheng Huang Australian National University EM Unit, RSBS Box 475, ACT 2601 Canberra, Australia Phone: 61-2- 6125-6553 Fax: 61-2- 6125-3218 http://www.anu.edu.au/EMU/
Is your cooling water cool on return? How long since your dif pump was cleaned and recharged? Vacuum fluid can diminish over time and when its down to half of the initial charge you may have that type of problem. Fluid will work well in some TEMs for years, but suspect the pump if its more than two years since the last fluid change - and less if the pump lived through a couple of mishaps. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, October 18, 2001 5:30 AM, Lilly M. [SMTP:lilly-at-cycy.net] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone, } } We are having a problem with the high vacuum on our HItachi H600 TEM. When } the microscope is } first turned on, the high vacuum is achieved but about 5 to 10 minutes later } it starts } dropping. The problem developed over time. In the beginning, it would drop } by very little and } go back up again. Later on, it started dropping by a lot more and it took } longer to get back } up again. Now, the vacuum is almost competely lost. Does anybody know what } might be causing } this problem? Any tips or suggestions? We appreciate the help and thank you } all in advance.
My colleague Dr. I. Pencea has a proposal for an aplication, if someone is interested in this please contact us.
Best regards, Prof. D. Bojin
" Dear Sir/Madam,
My name is Ion Pencea and I am Assoc. Professor with "Politehnica " of Bucharest, Romania, Materials Science & Engineering Faculty. My research field is materials characterization by WAXD, SAXD and XPS. I have gained a scholarship for training in microscopy investigations (SEM, TEM, AFM, etc.). The training period is of least 6 month and maximum 12 month depending on host agreement. Thus, I am looking for a laboratory which could accept me as a visiting scholar or other position (post doc., etc.). I have some experience in SEM and TEM with a TESLA microscopes (BS 350, BS 540) but I am interested in a modern SEM with EDS / WXD attachments, and of course in TEM and AFM. During this training period I can help with investigation on materials of host interest and other specific work (documentation, papers, reports, etc.). My scholarship will cover my accommodation but if it is possible to get work permission it will be perfect. If you are interested in my application, please contact us.
Most Hitachi microscopes have a seleniod valve that "vents" air into the vacuum line when the SEM/TEM is turned off. This prevents any residual vacuum from sucking oil back into the forlines and causing startup problems.
This seleniod valves get really hot during operation and, over the years, causes the hose to crack. This small crack allows air into the foreline thereby raising the foreline pressure & ultimately the SEM/TEM pressure. As the crack gets larger, the foreline pressure is worse, etc.
The first thing to do is to identify the Selenoid valve(s) in question. Trace the vacuum lines from the rotary pumps to a "T" in the line. The "T" should have a 3/8 in (6mm) OD rubber hose coming from the "T" to the top of the solenoid valve. The solenoid valve will be very hot. Either replace the hose or cut it shorter depending upon it's condition.
Hope this Helps.
Earl Weltmer Scanservice Corp.
----- Original Message ----- } From: "Lilly M." {lilly-at-cycy.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, October 17, 2001 12:30 PM
Judi Bowden writes ...
} ... } Do you see a link between science and art? If so, how and why? } ...
I think you are mixing apples and oranges. "Science" is a process by which we learn ... "art" is something we create and/or observe. However, that is not to say we cannot use a scientific process to learn how to create, or create differently.
We have examined numersous brain samples, both biopsy and autopsy material. The damage you will see due to autolysis will totally obliterate any difference you might have seen due to the use of formalin versus or prior to glutaraledehyde. Length of time storaged in fix will also affect the tissue less that length of time the person was dead before the tissue went into fixative.
The best ultrastructure would be obtained on biopsy material (hard to get), and the next best structure would be from rapid autopsies, like those done in 30 min after Alzheimer's Disease patients have had died for the special studies some folks are doing on rapidly decaying processes.
If one is comparing ultrastructure of one case to another, (s)he needs to note carefully the length of time the person was dead before the tissue went into fix (or the person was embalmed). This time can vary widely.
Sara Miller
On Thu, 18 Oct 2001, Allan Mitchell wrote:
} Date: Thu, 18 Oct 2001 12:07:18 +1300 } From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} } To: microscopy-at-sparc5.microscopy.com } Subject: TEM of cadaver tissue } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Kia Ora all } } We have a student starting to work with human cerebral blood vessels } obtained from cadavers. } } A literature search has revealed little information on working with } cadaver material at the TEM level. } } The questions from this student are; } } 1. Does anyone know of any publications resulting from TEM } investigations of samples from human cadavers, in particular cerebral } blood vessels and brain / nervous tissue ? } } } 2. The standard EM fixation texts talk about not using formalin to } fix for TEM and emphasize speed of primary fixation. My initial } results indicate that the ultrastructure of the cadaver cerebral } blood vessels has remained unexpectedly intact. } } Has anyone else experienced good ultrastructural results in the TEM } with cadaver material, embalmed in 10% Neutral Buffered Formalin and } after an initial post mortem delay of up to 24 hours ? } } } - What ultrastructural changes (at the organelle level) did you } expect and then what actual ultrastructural changes did you see ? } } } 3. What ultrastructural changes would you expect from such material } if it has been in 10% Neutral Buffered Formalin for up to 6 months } prior to processing and examination ? } } On behalf of our student, thanks for your help. } } Allan } } } -- } ------------------------------------------------- } Allan Mitchell } Technical Manager } Otago Electron Microscope Centre } C/-Department of Anatomy and Structural Biology } School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand } } Phone (03) 479 5642 or 479 7301 } Fax (03) 479 7254 } } Unit: http://www.otago.ac.nz/anatomy/emunit/ } Department: http://anatomy.otago.ac.nz/ } } } } "Life is a gift, don't waste it" } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Judy, For most of us, it isn't good enough just to take a picture that shows what we need to see, it also has to look good. I think that is where the art comes in. I've had 3 cover micrographs for the Scanning Journal (www.scanning-fams.org).Most of the images were from my Ph. D. research, but add some color...and you have ART!
Dr.Tina S. Schwach, President Microscopy Consulting Services Inc. 651-681-0112
----- Original Message ----- } From: {"01151938-at-mrc.vic.edu.au"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 16, 2001 6:09 PM
I heard a definition of art when I was in photography school many years ago. I wish I knew the source...
Art is anything, that when abstracted from its surroundings, embibes its viewer with a hieghtened or altered sense of thier own existence.
Soon after is when I went from fine art to scientific imaging because it does this so magically.
If you perform clinical TEM or SEM, I need your input on a College of American Pathology (CAP) reevaluation of the labor time they allot for these procedures. If you're willing to help, please contact me offline, and I will send you a draft of the operating procedures for your comments.
The CAP has CPT codes that determine how much hospital labs can be reimbursed. The descriptions for TEM and SEM procedures are out of date and appear to have been written by a lab director who was not actually an electron microscopist as some of the steps don't make sense. I have been asked by CAP to submit updated lists for consideration for adoption.
We prepare several hundred EM samples per year for Duke hospital, and I have, with the help of my 5 superb techs, listed each step and the time it takes to perform them. If you work in a hospital lab or are contracted to perform EM on human tissue, I would very much appreciate your input. Please contact me.
Thanks in advance, Sara
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Dear Lilly, The two things that I know that can cause that are: 1. the heater on the diffusion pump is not quite heating hot enough to boil the oil, 2. the diffusion pump oil is almost gone. The fact that you achieve high vacuum means that your system is almost working and has no leaks. At 12:30 PM 10/17/01 -0700, you wrote: } } Hi everyone, } } We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is } first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts } dropping. The problem developed over time. In the beginning, it would drop by very little and } go back up again. Later on, it started dropping by a lot more and it took longer to get back } up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing } this problem? Any tips or suggestions? We appreciate the help and thank you all in advance. } } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I have a few opinions about this topic since I am a research microscopist by day and an artist by night. One of my interests involves utilizing elements from science in the production of art. I have an exhibition/performance November 8th--24th at SomArts Gallery in San Francisco that will include some exemplary pieces. You are invited! You can learn more from the web site at http://SaintRubidium.com I will be adding additional descriptions of the art work to the web site before November 1st. Some examples are: a screen made of 6000 Petri dishes; micro art viewed with a light microscope and made with drosophila wings, human blood and copper sulphate; a video of a 30 foot by 30 foot array of data tapes from electrophysiology potassium channel recordings, an eight foot cylinder "totem" made from science journal cover photos; another totem made from radiographic films and transilluminated; a computer controlled 3-slide-projector with stereo sound presentation of botanical scanning electron micrographs made by Douglas Frank in Portland, Oregon.
I find the world of science rich with imagery and fascinating materials that are exciting to use in a manner that brings a different perspective on science to the public while investingating personal issues as well as some issues common to all of humankind and the universe. There are numerous people interested in the intersection of art and science. YLEM just celebrated their 20th anniversary with an excellent exhibit in San Francisco. More information can be found at http://www.ylem.org See their links page for connections to other people and organizations such as the San Francisco Exploratorium. Another resource is the journal Leonardo / the International Society for the Arts,Sciences and Technology. See: http://mitpress2.mit.edu/e-journals/Leonardo/index.html
I hope that these few words explain a little bit of the how and why of the connection between art and science. I must add, however, that I learned early on in my career that there are scientists that find this whole art interest in science repugnant. I'm still not clear why but can only speculate. I also find that the influence of my art on my science is much less clear to me except that it provides some motivation to survive the trudgery that experiments often require. Larry
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The American Chemical Society celebrates National Chemistry Week during November 4-10, 2001. This year's theme is "Celebrating Chemistry and Art." For more information, refer to http://www.acs.org and scroll down to National Chemistry Week.
Kristine Danowski Analytical Sciences Dow Chemical Company Midland, MI 48667 kldanowski-at-dow.com
} Hello: } Could somebody please define for me "home brew and ASR rules"? Does this } phrase refer to how reagents or antibodies are made, in-house vs. } commercially? } Thanks, } Steve Saul } } e-mail: ssaul-at-optronics.com
Dear Tom, I have prepared cell pellets for virus-infected cells many times. If the cells are in 6-well plates, one can process them in the plates thru the dehydration and release the cells (quickly!) with washes of propylene oxide. The cells can then be infiltrated in a P.O. resistent polyproylene test tube and spin down after each change of resin. When embedding, then pipet off the resin to the pellet and tranfer the pellet to beem capsules. Spin them in a piggy-back arrangement with the beem capsule tucked in a microcentrifuge tube. (Cut the microcentrifuge tube so the beem capsule fits) Close the cap, spin and top off with resin. If the cells are in suspension, one must spin after each step of fixation, dehydration, and infiltration before transfering them to beem capsules to embedd. But, you may process suspension cells in the polypropylene tubes which are easy to spin.
Barbara Plowman Univ. of the Pacific School of Dentistry 2155 Webster St. Rm 632 San Francisco, CA 94115 email: Bplowman-at-sf.uop.edu phone: 415-929-6692
I operate an H600 and have seen a similar problem. First, by turned "on" do you mean the high tension voltage (e.g.. 100kv) or do you mean physically starting the system from no pumps running to switching on the pumps and waiting for the DP's to heat up and pump the system down?
If you mean applying the 100kv, I've found that filling the large LN2 trap before switching on the 100kv virtually eliminates any vacuum swings.
The other problem may be that you need to clean the PENNING gauge vacuum sensor. if its heavily coated with junk it will short-out and you see it as vacuum drops and poor levels on the readout. There is a section in the H600 instrument manual that tells you how to clean the gauge. Its very easy to do and takes about 2-3 hours of your time. If needed please contact me off list and I can fax you the pages in the manual.
dz
-----Original Message----- } From: Lilly M. [mailto:lilly-at-cycy.net] Sent: Wednesday, October 17, 2001 3:30 PM To: Microscopy-at-sparc5.microscopy.com
Hi everyone,
We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts dropping. The problem developed over time. In the beginning, it would drop by very little and go back up again. Later on, it started dropping by a lot more and it took longer to get back up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing this problem? Any tips or suggestions? We appreciate the help and thank you all in advance.
} If you are interested in further exploring the connection, I suggest you } contact David Scharf of Los Angeles CA... David is an Artist who produces } color SEM photomicrographs and his work is outstanding.
The reason that I have an SEM is because of David Scharf. In 1976 I bought a copy of his book _Magnifications_ and was entranced by the images. Scharf clearly not only has mastery of the SEM, but also an artist's eye for composition (and a sense of humor, as well). As I looked through the images over and over, I came to believe that the SEM has to be one of the best toys ever invented. It took 25 years, but I now have my own toy.
I believe that science and art support each other very well. One of my other interests is pottery. My scientific background helps me understand which forms will be mechanically strong and which will be fragile, what chemicals in glazes do, how heat in the kiln works, how to conduct meaningful experiments... I feel sorry for those who attempt to create pottery _without_ the benefit of a scientific background. They are at a distinct disadvantage.
Scientific endeavors can also benefit from art. My favorite example is (was) O'Shaughnessy dam on the Scioto river near Columbus Ohio. I say "was" because the original dam no longer exists. The dam was beautiful. It was not just a rectangular block of concrete poured into the path of a river. It had graceful curves and portions of it were terraced into the surrounding rock. Rather than simply plummeting down a abrupt spillway, the water broke over a curved edge, cascaded down the main portion of the breast, and was reintroduced to the river with a reverse curve at the bottom. As a result, the water fell almost silently. More sound came from the small terraced portions, where the water might fall 5 or 10 feet at a time, than from the main spillway. O'Shaughnessy served its purpose well as a dam and was beautiful at the same time.
Hi Judy, There was a great talk this year at the M&M Meeting in Long Beach, CA related to that subject. The MSA Presidential Happenings lecture was presented by Stuart S. Sumida from California State University San Bernardino: From Bones and Stones to Balloons and 'Toons: Bringing Biology and Art Together in Realistically Animated Films. He's a paleontolgist and he has advised Disney animators on animal movement for such films as The Lion King, Mulan, etc. He would be a great person to contact. Perhaps someone has his email address?
On a personal note, I enjoy doing 35mm B&W macro photography. Last year I entered photographs in a juried show. Up to 3 photos could be entered, so I submitted 2 of my favorite 35mm photos and for a laugh I threw in a SEM of pollen grains on a Bumblebee's back. Needless to say I was truly miffed when the SEM was accepted and my others were rejected - ha! Anyway, it made me think about my work and how I approach it. I do try to frame things up in the most esthetically pleasing way and still have it contain all the scientific information that's needed. Even though I've separated my work from my art for many years, I think all microscopists are artists. Is anyone else observing our world so closely? ...well, besides the media;-) good luck with your project, Beth
} Dear Listers } I am presently undetaking an Honours degree at La Trobe University, } Victoria, Australia.I have been researching flower pollen using a Scanning } Electron and Confocal microscopes. } I am particularly interested to hear your views on the following question: } Do you see a link between science and art? If so, how and why? } Thank you for considering my question. } Yours sincerely } Judi Bowden } La Trobe University } Victoria } Australia
****************************************************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) ******************************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull. Aqualung
"Do you see a link between science and art? If so, how and why?"
Judi,
Linking creativity in art and science:
Creativity in art is not substantially different from creativity in science. Creativity in art/science is not a person suddenly struck by a blinding flash of inspiration/insight and then producing a masterpiece. Even the most radical artists/scientists start from classical knowledge and then spend years polishing their technique. [Picasso, Mozart, Einstein] (source: Julio Ottino, lecturer on "Creativity ? an Art View", an accomplished artist and Professor, and Chair, Department of Chemical Engineering, Northwestern University, Evanston, Illinois, USA).
"Even the most talented persons do not reach world class performance in their fields until they have devoted about 10 years or more of rather single minded attention to becoming an expert". What chiefly distinguishes creative thinking from more mundane forms [in art/science] are Ambiguity, Persistence and Knowledge - (i) willingness to accept vaguely defined problem statements and gradually structuring them, (ii) continuing preoccupation with problems over considerable periods of time, and (iii) extensive background knowledge in relevant and potentially relevant areas. (source: H. Simon, Nat'l Acad Sci Proc, 80, 4569-4571, 1983).
Art is the selection from a great number of variables (source: book, Zen and the Art of Motorcycle Maintenance). There are also a great number of variables to select from at the start of a new science project.
Regards,
Ev Osten
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000 U.S.A.
just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
From my background in physics I find this question very intriguing.
Art stands out by its resonance with our experience or by harmony with something within us. We marvel at the expression. Sometimes it expreses a play on patterns with nothing to do with function like designs on ancient potery while other times it's the expression of functional form, the bowmans weapon or the flying gull. We identify, at once the ideal human form in the Greek sculptures and dynamic figures from the caves in France while we see the vision and mastery of the artist.
With science it may not be so different. We paint with a bigger brush with a handle of technology, bristles of training, and paint of intellegent abstractions on a canvas of hypothetical vision. Does not this or that stroke of the brush resemble the observation? If not quite, then improve the stroke, refine the correspondance. Generalize the circle to an ellipse. Now the planetary cycles correspond... The Painting is never done. Maybe it is more like an evolving building with not so many master archetects and many builder craftsmen all who live in the structure they build daily. Their dedicated methodology is required to keep the structure sound with each new change.
But science is an art. It is an art of knowing, a craft of many participants. Mere scientific method cannot make breakthrough discoveries. It is an art of creative nurturing of insight correctly applied to each step.
The same honesty requred in the scientific method may also be required by any artist to acheive her mastery of technique.. The same aesthetic experience of the artist and viewer may also be part of the experience of scientist and student.
My $00.04 worth,
Dave -- David Barnard Wadsworth Ctr NYS Dept Health Albany NY 12201-0509 barnard-at-wadsworth.org 518 473-5299 voice 518 474-7992 fax
I am trying to do some SEM on nematodes with elaborate structures on their front ends. Unfortunately, these critters secrete some sort of sticky residue around these structures that makes it difficult to get good, clean specimens. The "stuff"is likely some sort of glycoprotein goo and I should (hopefully) be able to rinse it off with something, without damaging the integrity of the collagenous cuticle and without giving me other sorts of artifacts. Any ideas? -- ______________________________________________ Dan Bumbarger Graduate Student Depts. of Biology and Nematology University of California, Riverside Riverside, CA 92521 ______________________________________________
"With a little more deliberation in choice of pursuits, all men would perhaps become essentially students and observers, for certainly their nature and destiny are interesting to all alike. In accumulating property for ourselves or our posterity, in founding a family or a state, or acquiring fame even, we are mortal; but in dealing with truth we are immortal, and need fear no change nor accident."
Dear Michelle, Had the same problem when I was starting out. It turned out that I was alternating with someone who was using tungsten film and removed the daylight filter for his photography. On another occasion when I was the lone user, I found that there were lots of voltage changes during the day and photos taken at one time were different from those taken at another. The immediate way to handle the voltage problem is to check the power supply voltage level for every exposure. When I had a class taking photos, I used a tabular sign-in sheet with columns for: Power supply setting; Iris diaphragm open; field diaphragm adjusted; objective; time of photo; trinocular prism setting (1,2, or 3); etc. I think you get the point. If your line voltage is at fault, the only way you can prove it is to have someone perform a time study to record those changes over a 24hr period at your outlet. The next problem that I have seen is caused by picking a roll of film on Monday that is designated for use with a tungsten source and another roll on Thursday that is designated for use with daylight. Oh, yes! AND FINALLY, if your photo detector is set for spot metering rather than averaging, a small change in location can/will have a great effect on the result, which will appear to be unpredictable. From all of the above, your best bet is the proper compensating filter when using artificial light (LBD (Olympus) for daylight-type(5500K) and LBT for tungsten-type(3400K) films) For years I have treasured a book I acquired from Olympus entitled, "How To Improve Photography Through the Microscope" (Part # M132E-0886T). There is another that I acquired from Leitz before it became Leica.
Hope that one small part of this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: mryder-at-brookes.ac.uk } Sent: Wednesday, October 17, 2001 8:00 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist:LM color changes in photographs } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (mryder-at-brookes.ac.uk) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } October 17, 2001 at 05:31:30 } -------------------------------------------------------------------------- } - } } Email: mryder-at-brookes.ac.uk } Name: Michelle Ryder } } Organization: Oxford Brookes University } } Education: Graduate College } } Location: Headington Campus, Gipsy Lane, Oxford, OX3 0BP } } Question: Why do my wholemount lowish power root } photographs sometimes come out yellow, on the } same film, with the same lighting. Sometimes } the same root taken a few minutes later will } be yellow when it was the usual white on a previous frame. } } } } -------------------------------------------------------------------------- } - } }
"This method did not work in our hands." "I've been looking at histologic sections for 20 years, and I never made that connection." "How did you get these preparations to have such low background?" If there is no art in the application of the principles of science to the business of discovery, then there is no discovery.
Here's to the art(full) scientist who discovers what others have missed, because she sees the truth in corn while everyone else is treating it like food.
Regards to all,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
Lilly, I've seen this kind of behavior if the oil level in the diffusion pump gets low (I'm assuming that it is diffusion pumped). What happens is that the oil starts to boil and the DP pumps. When the oil is all boiled off, it stops pumping until some condensed oil runs down the side and back into the boiler section where it can boil and pump a little more.
The solution is to remove the DP, clean it and recharge it with the proper amount of the proper oil. Generally this problem occurs when you either have a large air leak or the DP is frequently vented when still quite hot, a definite no-no.
Ken Converse owner Quality Images third party SEM service Delta, PA
Lilly M. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi everyone, } } We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is } first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts } dropping. The problem developed over time. In the beginning, it would drop by very little and } go back up again. Later on, it started dropping by a lot more and it took longer to get back } up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing } this problem? Any tips or suggestions? We appreciate the help and thank you all in advance. } } }
Shaf, My wife will tell you art can be taught just like science. There are basic principles involved that anyone can learn. Just like science, though, some have more "innate", "natural",( you choose the term) ability. This is why there are some outstanding artists and some outstanding scientists. The creativity is the difference between the norm and the great in both cases. The same holds in most every field of endeavor that I can think of.
Ken Converse
michael shaffer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Judi Bowden writes ... } } } ... } } Do you see a link between science and art? If so, how and why? } } ... } } } I think you are mixing apples and oranges. "Science" is a process by } which we learn ... "art" is something we create and/or observe. However, } that is not to say we cannot use a scientific process to learn how to } create, or create differently. } } my 0.02 ... shAf :o) } } } }
I've looked at numerous post mortem retina specimens. I agree with Sara, though do find that glut fixation is better than form, even though there are P/M effects to consider. Unfortunately it's not always possible to say that a tissue from, say, 8 hours P/M will be better than one from, say 12 hours. Other variables, such as temperature of the body during that time come into play. Sometimes 8 (or 12 or even 15) hours P/M is fine, sometimes useless. Obviously however, less is generally better. Some cells seem a lot more "fragile" than others and deteriorate very rapidly - it's useful to find, for your tissue, which these are and use them as a rough measure of P/M effects. Blood vessels do seem pretty tough and I've seen quite good vessels when surrounding tissue is a mess. I haven't had to embed much for EM after long term storage so can't answer question 3, but have done immuno work on form fixed material up to years old and it was fine for LM. As for what U/S changes to expect in P/M material, that's a question that could take up a whole book! Just look carefully and compare your own specimens with each other and with what you know to be good U/S. It's all fairly obvious with experience.
Diana
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Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Yes it is possible to check electronic devices. From our experience it may give better results if you are applying a load (mechanical and/or thermal) to the device or much better to the entire pcb. Sometimes these tiny cracks are occurring only if the load is applied.
email: mklein-at-visitec-em.de WWW: http://www.visitec-em.de * Home of world's largest SEM *
-----Ursprüngliche Nachricht----- Von: Mark Riggs [mailto:Mark.Riggs-at-asml.com] Gesendet: Donnerstag, 18. Oktober 2001 22:11 An: microscopy-at-sparc5.microscopy.com Betreff: semming a capacitor
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just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
} } } } "Do you see a link between science and art? If so, how and why?" } } Judi, } } Linking creativity in art and science: } } Creativity in art is not substantially different from creativity in } science. Creativity in art/science is not a person suddenly struck by a } blinding flash of inspiration/insight and then producing a masterpiece. } Even the most radical artists/scientists start from classical knowledge and } then spend years polishing their technique. [Picasso, Mozart, Einstein] } (source: Julio Ottino, lecturer on "Creativity ? an Art View", an } accomplished artist and Professor, and Chair, Department of Chemical } Engineering, Northwestern University, Evanston, Illinois, USA). } } "Even the most talented persons do not reach world class performance in } their fields until they have devoted about 10 years or more of rather } single minded attention to becoming an expert". What chiefly distinguishes } creative thinking from more mundane forms [in art/science] are Ambiguity, } Persistence and Knowledge - (i) willingness to accept vaguely defined } problem statements and gradually structuring them, (ii) continuing } preoccupation with problems over considerable periods of time, and (iii) } extensive background knowledge in relevant and potentially relevant areas. } (source: H. Simon, Nat'l Acad Sci Proc, 80, 4569-4571, 1983). } } Art is the selection from a great number of variables (source: book, Zen } and the Art of Motorcycle Maintenance). There are also a great number of } variables to select from at the start of a new science project. } } Creativity is not bounded by art or science. The closer you get to the leading edge the more science becomes an art. The only difference is in art you have critics and in science you face peer review.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
Hi Dan, We used to remove the stylet secretions from rot-knot and soybean cyst juveniles (very sticky stuff) by adding 1 part of a high pH buffer (0.1 M Tris-NaOH, somewhere around pH 11-12) for 2 minutes or so. Of course, we were attempting to collect the secretions and not preserve the nematode. Although they seemed to be fine, as we could usually do this for several days after washing the 'tode. John Shields EM Lab University of Georgia
On 18 Oct 2001, at 14:54, Dan Bumbarger wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I am trying to do some SEM on nematodes with elaborate structures on } their front ends. Unfortunately, these critters secrete some sort of } sticky residue around these structures that makes it difficult to get } good, clean specimens. The "stuff"is likely some sort of glycoprotein } goo and I should (hopefully) be able to rinse it off with something, } without damaging the integrity of the collagenous cuticle and without } giving me other sorts of artifacts. Any ideas? -- } ______________________________________________ Dan Bumbarger Graduate } Student Depts. of Biology and Nematology University of California, } Riverside Riverside, CA 92521 } ______________________________________________ } } } "With a little more deliberation in choice of pursuits, all men would } perhaps become essentially students and observers, for certainly their } nature and destiny are interesting to all alike. In accumulating } property for ourselves or our posterity, in } founding a family or a state, or acquiring fame even, we are } mortal; but in dealing with truth we are immortal, and need fear no } change nor accident." } } } -H.D. Thoreau, "Walden" }
Well, yes and no. I agree that creativity is basal to both art and science, but surely the two are fundamentally distinct at least as far as their aims are concerned. The aim of science is to discover the truth (whatever that may be! - see e.g. Popper) about the natural world and its properties. Scientists use their creative imagination to devise ways of discovering facts about the world. Artists use their creative imagination to create objects -artefacts- which never have been of this world before. Scientists also create artefacts, but usually blush when one is pointed out to them. I find it fascinating that the most imaginative scientists are also profoundly aware of the arts, and are frequently very accomplished artists. Equally many artists are intrigued by the findings of science, if not actually by the methods, so the possibility exists of considerable common ground between the two activities as far as the mental / intellectual processes are concerned. But the key features that distinguish science from art are the accumulation and evaluation of information, the use quantitative analysis and of logic, and the application of a rigorous scientific methodology to distinguish fact from non-fact. None of these things are fundamental to art, indeed with a few exceptions their use seems to be at best optional, if not scorned altogether. In other words it is the prerogative of the artist to let the mind be totally free of the contraints of reality. Scientists do not have that luxury.
Chris
} Creativity is not bounded by art or science. The closer you get to the } leading edge the more science becomes an art. The only difference is in art } you have critics and in science you face peer review. } } Gordon } } Gordon Couger } Stillwater, OK } www.couger.com/gcouger } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
when was the last time your DP oil was replaced? It could be 'cracked' or, as was explained to me by a JEOL engineer, saturated by trying to remove more air from the column than it was capable due to a variety of reasons like too short a prepump(roughing) time, and/or improper valving sequences from rough pumping to high vac or a small column leak.
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Hi everyone,
We are having a problem with the high vacuum on our HItachi H600 TEM. When the microscope is first turned on, the high vacuum is achieved but about 5 to 10 minutes later it starts dropping. The problem developed over time. In the beginning, it would drop by very little and go back up again. Later on, it started dropping by a lot more and it took longer to get back up again. Now, the vacuum is almost competely lost. Does anybody know what might be causing this problem? Any tips or suggestions? We appreciate the help and thank you all in advance.
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Hello semmers, Since I have had such great help in the past from all of you, I have decided to once again seek information from this great source. I am wondering if any of you use reactive ion etching, or even more helpful would ICP-RIE. If so, it would be great if you could answer some questions for me. Please contact me off line, as it isn't really a microscopy question. Thanks again Nick
Nicol Aitken Sample Preparation and Imaging Specialist Research and Development Semiconductor Insights Inc. email:nicol-at-semiconductor.com (613)599-6500 ext.4300
a week ago I wrote asking about a problem related to image distortion on our SEM. Thanks to all of you who wrote suggestions to us. the technicians are studying the solution of the problem.
My question now is related to sample preparation of paper for studying the thickness of the fibers. The idea is to obtain cross sections of paper sheets showing the sections of the fibers. How would you prepare it in order to avoid swelling or deformation of the sections? Embedding would not be the ideal method since the surface of the same paper has to be observed.
Any suggestions will be very much appreciated.
Thanks a lot
Silvia Montoro CERIDE G|emes 3450 3000 Santa Fe - Argentina csedax-at-ceride.gov.ar
Mark: a lot depends on what type of capacitor it is. More detail about the cap would be helpful. In my experience, small ceramic surface-mount caps are more prone to problems in the area than large metal-can electrolytics. I agree with Martin Klein that examining the cap in its native habitat would be the best, but it all depends on how big your SEM chamber is and where you suspect the cracks might be occurring. If they are between the cap and the board; they'll be really hard to see in the SEM. A word of advice: this is best done in a field-emission SEM or variable-pressure SEM because charging is going to be a big problem and I'm assuming you don't want to coat the board with gold or carbon.
Mark Riggs wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) SC Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
We have a Polaron Turbo Coater carbon evaporator, model E6700, probably 10 - 15 years old, which is experiencing some problems. In spite of acceptable vacuum and functioning pirani gauges, the penning gauge is not turning on, and one of the fuses is blowing every time we turn it on.
We don't have schematics for any of the boards in the instrument, just a schematic which gives an overview of the entire instrument, but no specifics of individual boards.
If anyone out there has schematics for any boards in this instrument, and would be willing to share this info, could you please contact me off line?
Thanks, Wharton Sinkler UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017
Hello, I'm trying to find a couple of recommended references for phytoplankton and other small items to be fixed/dried for SEM. The samples have the problem of getting lost during the CPD process. I have the following, but I cannot find the book Scanning Electron Microscopy II. Has anyone seen these references and are they correct as I have them:?
Preparation of Microbiological Specimens for Scanning Electron Microscopy, L.P. Watson, A.E. Mckee, and B.R. Merrell. Scanning Electron Microscopy/1980/II, pages 45-56.
Specimen Preparation Techniques for Aquatic Organisms, T.K. Maugel, D.B. Bonar, W.J. Creegan and E.B. Small. Scanning Electron Microscopy/1980/II, pages 57-77.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Dear Judy, To your question: Do you see a link between science and art? If so, how and why? The mediating event between the interiority and exteriority of our experience of the world is in the aesthetic moment according to Jamesian philosophy. It forges the union between past and future, between certitude and expectation. As many other have said previously in this discussion -especially David Barnard's excellent observation - their work often hinges on the awareness of beauty within the work. It ties together the craft of rigorous method with optimism of our theory.
I have spent thirty years working in photography in all its aspects, from street documentary to fashion studio work, from mug shots to gallery work. One of the most exciting bodies of work that I ever produced was as student of EM at the VA Center in Albany, NY, now sadly closed. I shot something like 4 times the amount of film than most students did. Why? Because the electron microscope, especially the TEM, is nothing more than a camera, but what a wonderful camera it is! It investigates a landscape, searches for a recognizable form, establishes proportions and makes measurements, always looking for 'significance' however that may be defined . In short, exactly the same thing I try to do in conventional landscape photography. It' s all a part of the attempt to make sense of the perceptible world, or to make the imperceptible visible and then intelligible.
I recommend to you the following: Art Forms in Nature, Ernst Haeckel and Karl Blossfeldt's Art Forms in the Plant World
Fred and Michelle, I'd like to add a few additional and perhaps unrelated points to getting consistent results. Having done a a lot! of photomicrography especially for publication in text books, another method for getting consistent results is to buy a block of film that is all from the same lot (check the labels). I also learned that film is made to have a known storage or shelf time until it reaches the optimum color balance, so you have to find out what that is as well. Finally, I had to find a processor that ran test strips at the beginning of the day to assure that the process was correct, then I made sure that they did my film first thing in the morning. Regarding filters, I had to buy a set of light balancing and color compensating (CC) filters to get "perfect" color balance. Naturally, I always bracketed by about 1 stop in 1/3 stop increments as my time was more expensive than the film. I learned that the voltage fluctuated slightly (dropped) and had to turn the lamp on and wait about 30 min to get it stable. Obviously I was doing very critical color work and this all may be overkill for you. Regards from an oldtimer whose using digital now :-) Damian } } Dear Michelle, } Had the same problem when I was starting out. It turned out that I } was alternating with someone who was using tungsten film and removed the } daylight filter for his photography. On another occasion when I was the } lone user, I found that there were lots of voltage changes during the day } and photos taken at one time were different from those taken at another. } The immediate way to handle the voltage problem is to check the power supply } voltage level for every exposure. } When I had a class taking photos, I used a tabular sign-in sheet } with columns for: Power supply setting; Iris diaphragm open; field } diaphragm adjusted; objective; time of photo; trinocular prism setting (1,2, } or 3); etc. I think you get the point. If your line voltage is at fault, } the only way you can prove it is to have someone perform a time study to } record those changes over a 24hr period at your outlet. } The next problem that I have seen is caused by picking a roll of } film on Monday that is designated for use with a tungsten source and another } roll on Thursday that is designated for use with daylight. } Oh, yes! AND FINALLY, if your photo detector is set for spot } metering rather than averaging, a small change in location can/will have a } great effect on the result, which will appear to be unpredictable. } From all of the above, your best bet is the proper compensating } filter when using artificial light (LBD (Olympus) for daylight-type(5500K) } and LBT for tungsten-type(3400K) films) } For years I have treasured a book I acquired from Olympus entitled, } "How To Improve Photography Through the Microscope" (Part # M132E-0886T). } There is another that I acquired from Leitz before it became Leica. } } Hope that one small part of this helps, } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } } } } ---------- } } From: mryder-at-brookes.ac.uk } } Sent: Wednesday, October 17, 2001 8:00 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Ask-A-Microscopist:LM color changes in photographs } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (mryder-at-brookes.ac.uk) from } } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } } October 17, 2001 at 05:31:30 } } -------------------------------------------------------------------------- } } - } } } } Email: mryder-at-brookes.ac.uk } } Name: Michelle Ryder } } } } Organization: Oxford Brookes University } } } } Education: Graduate College } } } } Location: Headington Campus, Gipsy Lane, Oxford, OX3 0BP } } } } Question: Why do my wholemount lowish power root } } photographs sometimes come out yellow, on the } } same film, with the same lighting. Sometimes } } the same root taken a few minutes later will } } be yellow when it was the usual white on a previous frame. } } } } } } } } -------------------------------------------------------------------------- } } - } } } } } }
Silvia Montoro wrote: ========================================================= ............... My question now is related to sample preparation of paper for studying the thickness of the fibers. The idea is to obtain cross sections of paper sheets showing the sections of the fibers. How would you prepare it in order to avoid swelling or deformation of the sections? Embedding would not be the ideal method since the surface of the same paper has to be observed.
Any suggestions will be very much appreciated. ========================================================== You are quite correct to be concerned about possible swelling and/or deformation of the paper structure if embedded. However, another concern is the possible swelling of the fibers themselves with the embedding resin, thereby giving you a diameter figure that is much larger than reality.
Our approach has been to gold coat both sides in a sputter coater before embedding. It is always a trial and error exercise to determine the "right" thickness of the gold layer, the purpose for which is to a) passivate individual fibers to reduce the tendency of the embedding resin to penetrate and swell individual fibers and b) reduce the "elasticity", or the ability of the larger structure to be changed by the embedding media. We find that Pt is somewhat better than gold because of its smaller grain size and the greater hardness results in a lowered tendency for the metal to "smear".
We have always had our best results with our own SPI-Pon™ 812 Epoxy Embedding Resin Kit but we believe some of the other Epon® "substitutes" would work just as well.
Using room temperature sectioning, you can then thin section the block and by TEM, the gold layer acts as a nice decoration line outlining the fibers. If there has been excessive fiber swelling, usually there are signs that this is happening, at least once one develops experience in looking at such micrographs. If you don't have a TEM, you can look by SEM at the "faced-off -piece", that is, the block from which the last section was taken. So long as you are using a thin layer of conductive metal, usually there is enough contrast from the gold decoration line to resolve well the individual fibers . Obviously this technique will work better with thinner than it will with thicker papers. It might not be the perfect solution but it does permit one to make measurements on the fiber diameters. Also, at times a short plasma etching with oxygen on the faced-off-surface will enhance further the contrast between the gold layer and the fibers and embedding resin.
If we would be doing this kind of work today, and wanting to look at the faced-off-piece by SEM, instead of coating with gold we would coat with 1 nm of osmium metal in the OPC Osmium Coater. Although a heavy metal, that thickness is thin enough that it does not interfere with being able to see contrast from the underlying gold (decoration) layer.
Note: You should be using a "materials science" rather than a life science (LS) diamond knife since they are cheaper than an LS knife and also, the inorganics in most papers would "kill" a LS knife for future use on LS samples.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
on 10/18/01 11:32 PM, Robert Underwood at underwoo-at-u.washington.edu wrote:
} } I heard a definition of art when I was in photography school many years } ago. I wish I knew the source... } } Art is anything, that when abstracted from its surroundings, embibes its } viewer with a hieghtened or altered sense of thier own existence. } Dear Robert and other listers, When I was an undergraduate, I was arrogant enough to propose a definition of art--one which would include such pieces as a number of bricks arranged in a neat pile (on display at MOMA). I defined art as the projection of the universe onto some chosen set of limitations. Good art enlightens the artist and audience about the nature of the universe by reducing it to an understandable piece; whereas bad art is unsuccessful in this enlightenment (so the value judgement rests with the audience). Not only do I still think that this is a valid definition, but I can also see how this applies very obviously to science. IMHO, therefore, science and art are not separate, but science is art. Yours, Bill Tivol
I'm not sure that this is off topic. Of course, Nestor can correct me if I am wrong.
I do RIE and ICP-RIE etching of IC passivation followed by SEM imaging on a somewhat routine basis. ICP is a new method and is quite improved over traditional RIE methods. Passivation removal is quite difficult based on power, pressure and gasses. For oxide, I use CF4 and O2 at a pressure of about 10mT. I'm not certain of the power.
My particular work in this area is to SEM image ICs which have been de-passivated. RIE and ICP-RIE accomplish this in different final outcomes. I find dramatic differences between packaged die and bare die.
gary g.
At 09:39 AM 10/19/2001, you wrote:
} Hello semmers, } Since I have had such great help in the past from all of you, I have } decided to once again seek information from this great source. I am } wondering if any of you use reactive ion etching, or even more helpful would } ICP-RIE. If so, it would be great if you could answer some questions for me. } Please contact me off line, as it isn't really a microscopy question. } Thanks again } Nick } } } } Nicol Aitken } Sample Preparation and Imaging Specialist } Research and Development } Semiconductor Insights Inc. } email:nicol-at-semiconductor.com } (613)599-6500 ext.4300
Dear All: Thank you for those who replied to my posting regarding anti-amylase antibody search. I eventually located one at the Chemicon. Now, I am looking for a polyclonal anti-synaptophysin antibody. I know that there are many companies carry this product. But I would like to know users' experiences before I decide on which one to purchase from. If you have used this antibody from a particular company, would you share the following information with me. Thank you a million in advance.
Supplier name: Samples labeled: (cell culture or tissue type, species...) Labeling type: (immuno fluorescence, immunoperoxidase, or immunogold?) Labeling procedure: (pre-embedding or post-embedding?) Fixative used: Embedding method used:
By the way, for those who do not have Linscott's directory, Try www.sciquest.com. I do my antibody search on this site. It is very easy to use.
The type of capacitor determines what you have to do. Titanate type chip capacitors consist of alternating metal electrode sheets sandwiched between the dielectric layers, both of which are laid down by printing methods and then fired into a monolithic structure. End metallization is applied so that the alternate protruding plate layers are electrically joined to a metal coating on each end. It is a robust structure but because it is made of very hard, rigid material, locating an internal fracture can be difficult. A fracture that extends to the outside can usually be seen in the SEM without coating the specimen. The tendency for dielectrics like titanates to charge is highly dependent on the orientation of the subject to the beam. One can usually tilt the faces of the capacitor to find an angle where charging is not a problem and then simply search for the crack. Using the backscatter detector instead of the secondary electron detector will also help to minimize charging interference.
Apart from the SEM, a fluorescent dye can also be used for an exterior crack. Magniflux makes such compositions but you can find others by doing a search with the terms "dye penetrant" and "non-destructive testing". The usual procedure is to place the cap in the dye inside a small vessel in which the pressure can be reduced. The vessel is partially evacuated so that upon return to room pressure the dye is drawn into the crack. The cap is then quickly rinsed and observed under a ultraviolet light with a low power microscope. Dye tends to creep out of the crack revealing its position.
Internal cracks can take different forms. One problem type is where the end metallization separates from the edge of one or more of the plates. This is a common source of temperature dependent, intermittent faults. In this case some destructive sample prep is required and there is a chance that the defect will be lost or "healed" in the course of the prep. One can determine the edge direction of the plates and then, using metallographic methods, grind and polish off the exterior ceramic coating to expose the edges of the plates. Don't go beyond the minimum necessary to expose the edges. Use a nap-less cloth for polishing, very light pressure and diamond abrasive to minimize smearing of the metallization. The point where each plate intersects it's respective end cap is a critical area to check for discontinuities. In the SEM you can now use the secondary detector and some charging to your advantage by grounding one endcap and leaving the other electrically floating. A plate that is charging up while its companions are not reveals where a discontinuity exists. Conversely, a plate that fails to charge when its companions on the same endcap are doing so reveals a short to the other endcap. Simple cracks in the internal dielectric can also be revealed by orienting the subject to where charge accumulates and edge effects accentuate the charging. If your SEM is set up for specimen current imaging, compare what you see in this mode with the results of SEI and BEI.
Good luck.
John Twilley Conservation Scientist
Mark Riggs wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } just heard a phone message inquiry about the ability of a sem to evaluate "micro-cracking" of a capacitor's outer shell. the requestor believes the "micro-cracking" might be indicative/source of performance problems. is this possible, and how do i sem it? mr
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Morning Dan, One might suggest warm physiological saline, but some will recommend you try what one of my children called 'goozyme', three others called 'spit', or I call, saliva, but most will recommend amylase, or some other -ase, which will cost unless you find some in a nearby refrigerator. While there are some who will interrupt this conversation by mentioning that the former -ase (that is, amyl-), only hydrolyzes glocogen, I have seen it do nicely on what used to be called mucous secretions, if your source of amylase is saliva (p.c.?). NOTE1: All protein-goo is preserved (cross-linked) by fixation longer than necessary to terminate (p.c.?) the subject (p.c.?). If it must be removed, then one should try to do it prior to fixation - not necessarily prior to termination (p.c.?). NOTE2: The above methods can be tried in any sequence for good or ill, but be forewarned that your actions will be misunderstood if you are seen by colleagues to be expectorating (p.c.?) at a worm. NOTE3: Of course, if you continue to work with worms with rhinitis, you must suffer the consequences. In all seriousness, however, if one's worm has a cold, could one preempt the goo with oxymetazoline hydrochloride. This problem, of course, will remind us all that one cannot grasp a live hagfish [old or young!], because it ties itself in a knot, secretes a bucket of goo, and then extricates itself by moving the knot (a posteriori), all of which hasn't much to do with your goo Dan, but does present a much more messy problem that others [I know not who!] have had, which should make you feel better about your, only slightly, goo'd animal. It is, after all, Monday morning, and having seen our grandson off on Saturday, Grandma and Grumpy spent the rest of the weekend, unsuccessfully trying to catch up on sleep. As always, we hope that if the above doesn't bring immediate ease of the problem, it will, at least engender a smile.
Regards and commiseration to all those who have wormy problems,
Fred Monson
(p.c.) = (polite conversation?), of course!
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) So, I will answer the phone with my name, not the name of my workplace. West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Dan Bumbarger } Sent: Thursday, October 18, 2001 5:54 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Dirty Nematodes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying to do some SEM on nematodes with elaborate structures on } their front ends. Unfortunately, these critters secrete some sort of } sticky residue around these structures that makes it difficult to get } good, clean specimens. The "stuff"is likely some sort of glycoprotein } goo and I should (hopefully) be able to rinse it off with something, } without damaging the integrity of the collagenous cuticle and without } giving me other sorts of artifacts. Any ideas? } -- } ______________________________________________ } Dan Bumbarger } Graduate Student } Depts. of Biology and Nematology } University of California, Riverside } Riverside, CA 92521 } ______________________________________________ } } } "With a little more deliberation in choice of pursuits, all men would } perhaps become essentially students and observers, for certainly } their nature and destiny are interesting to all alike. In } accumulating property for ourselves or our posterity, in } founding a family or a state, or acquiring fame even, we are } mortal; but in dealing with truth we are immortal, and need fear no } change nor accident." } } } -H.D. Thoreau, "Walden" } } }
I realize this topic has been hammered out in this forum a few times (for everything except the Nikon Optiphot superwide trinocular) ......can someone direct me to any information that would be helpful? ("I can do this .......yeah!!!") Thanks,
Hello. I am a marine biology graduate student at Memorial University of Newfoundland, Canada. One of my research topic is to study the microstructure of the statolith in the pelagic marine tunicate (Larvacea), Oikopleura vanhoeffeni. The statolith is a small cylindrical calcium build up (10 micrometer diameter, 20 micrometer height), attached to the brain of the tunicate. This bony structure is contained in the membraneous statocyst and probably works as a gravity detector. I would like to isolate this structure for SEM and TEM analyses. After this, Xray microcroprobe and other procedures will follow.
Dissecting the brain is easy but taking out the statolith from the statocyst is the difficult part. Visualizing the statolith can be done by calcium specific stain, alizarin red. I have tried to dissect the statolith out by using fine insect pins ( { 20 micron tip size), but the pins are too large. Doing this under dissecting scope is impossible since 4x magnification does not allow me to do such microdissection.
I tried to dissolve the statocyst with bleach and hydrogen peroxide to isolate inorganic statolith. However, after this procedure, I can no longer visualize the statolith since alizarin red stain dissolves instantaneously in these chemicals.
Sonication was also done to rupture the statocyst and pop the statolith out but it did not work.
Do you have any suggestion on how I can separate this structure? Sincerely, Nami Choe ____________________________________________________________________________
Nami Choe Ocean Science Centre Memorial University of Newfoundland St. John's, NF A1C 5S7 Canada phone:(709)737-3247 fax:(709)737-3220
I don't know anything about the structure you describe, but check out these references:
Howell DN, Miller SE. Identification of viral infection by confocal microscopy. 1999. Methods in Enzymology. Academic Press, 307:573-91.
Miller SE, Levenson RM, Aldridge C, Hester S, Kenan DJ, Howell DN. 1997. Identification of focal viral infections by confocal microscopy for subsequent ultrastructural analysis. Ultrastruc Pathol 21:183-193.
Briefly, we make "vibratome" sections, stain the slices in propidium iodide, and examine the tissue by confocal microscopy for areas of interest. We also sometimes use antibodies with a fluorescent tag. We then cut out a smaller area including the structure of interest (1 x 1 mm) in a shape of the state of Nevada so that you will know if the section gets flipped over.
If your structure is something that you can recognize morphologically, and you can deal with a 100 to 400 um thick section, this might work.
Good luck,
Sara Miller
On Mon, 22 Oct 2001, Nami Choe wrote:
} Date: Mon, 22 Oct 2001 15:37:43 -0230 (NDT) } From: Nami Choe {c56nc-at-morgan.ucs.mun.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: microdissection before SEM/TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopy members; } } Hello. I am a marine biology graduate student at Memorial University of } Newfoundland, Canada. One of my research topic is to study the } microstructure of the statolith in the pelagic marine tunicate (Larvacea), } Oikopleura vanhoeffeni. The statolith is a small cylindrical calcium } build up (10 micrometer diameter, 20 micrometer height), attached to the } brain of the tunicate. This bony structure is contained in the } membraneous statocyst and probably works as a gravity detector. } I would like to isolate this structure for SEM and TEM analyses. After } this, Xray microcroprobe and other procedures will follow. } } Dissecting the brain is easy but taking out the statolith from the } statocyst is the difficult part. Visualizing the statolith can be done by } calcium specific stain, alizarin red. I have tried to dissect the } statolith out by using fine insect pins ( { 20 micron tip size), but the } pins are too large. Doing this under dissecting scope is impossible since } 4x magnification does not allow me to do such microdissection. } } I tried to dissolve the statocyst with bleach and hydrogen peroxide to } isolate inorganic statolith. However, after this procedure, I can no } longer visualize the statolith since alizarin red stain dissolves } instantaneously in these chemicals. } } Sonication was also done to rupture the statocyst and pop the statolith } out but it did not work. } } Do you have any suggestion on how I can separate this structure? } Sincerely, } Nami Choe } ____________________________________________________________________________ } } Nami Choe } Ocean Science Centre } Memorial University of Newfoundland } St. John's, NF } A1C 5S7 Canada } phone:(709)737-3247 } fax:(709)737-3220 } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
From root Mon Oct 22 17:12:25 2001 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA28070 for dist-Microscopy; Mon, 22 Oct 2001 16:34:28 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA28067 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 22 Oct 2001 16:33:57 -0500 (CDT) Received: from renko.ucs.ed.ac.uk (renko.ucs.ed.ac.uk [129.215.13.3]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA28060 for {microscopy-at-sparc5.microscopy.com} ; Mon, 22 Oct 2001 16:33:46 -0500 (CDT) Received: from MESH900 (dialup-169.publab.ed.ac.uk [129.215.38.169]) by renko.ucs.ed.ac.uk (8.8.7/8.8.7) with SMTP id WAA02642; Mon, 22 Oct 2001 22:30:18 +0100 (BST) Message-ID: {000d01c15b41$4cf1c740$a926d781-at-ed.ac.uk} Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk}
Nami My guess is you are probably already on the right tack with your attempts to isolate the inorganic statoliths by treatment with hypochlorite and peroxide. If you next wash these reagents out of your preparation, Alizarin red will again stain the statoliths, so you can see them. The main problem may be to handle the statoliths during these treatments. One effective way of containing such small objects through a series of reactions or fixation/dehydration steps is to contain them in a Millepore filter unit, exchanging the solutions using a disposable syringe. There is a wide range of filter media with different chemical compositions and pore sizes to choose from. The majority will retain object in the size range of your statoliths.
hope this helps Chris Jeffree
----- Original Message ----- } From: "Nami Choe" {c56nc-at-morgan.ucs.mun.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, October 22, 2001 7:07 PM
to all, My question concerns connecting a light source to a metallograph. I have inherited a Zeiss metallograph with a mismatched light source, Optiquip. The unit arrived without a collar connecting the lamp to the metallograph. The equipment is consists of Zeiss metallograph (gray), 47 17 65 Optiquip light source (creme), model 770. I would appreciate comments from anyone having experience connecting these 2 components or knows of a supplier that can assist. Apparently, the length of the collar is critical and perhaps the diameter.
We have mounting solutions for the Coolpix cameras to fit most microscopes through a C-mount, phototube or eyepiece tube. We have the cameras and accessories as well. A number of configurations are available depending on the microscope. I would be happy to assist in finding the proper mounting, for most instruments we need brand and model number to determine requirements.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
To: Great Minds in the Halls of Science Wisdom!
I realize this topic has been hammered out in this forum a few times (for everything except the Nikon Optiphot superwide trinocular) ......can someone direct me to any information that would be helpful? ("I can do this .......yeah!!!") Thanks,
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (charlted-at-mcmaster.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, October 22, 2001 at 12:59:02 ---------------------------------------------------------------------------
Question: I am wanting to find out better embedding techniques for light microscopy which will allow me to remove a lot of the chlorophyll present in the plant tissue but retain as much anthocyanin as possible. I am currently trying to determine anthocyanin location in a particular plant leaf. My first embedding process involved using a 5% gluteraldehyde and phosphate buffer, followed by a dehydration in increasing concentrations of ethanol and then embedding using JB-4 epoxy resin kit. However, no anthocyanin is present after the above process. Do you have any ideas of how I might retain the anthocyanin, reduce the chlorophyll and observe the pigment location using a light microscope. EM will be used at a later date, but for now I am curious on how do do the above process. Thanks for your help, Emily Charlton, McMaster University, CANADA
The Department of Biological Sciences at SUNY-Brockport is searching for a Chairman. The vacancy announcement follows. We have an Hitachi 7000, several ulytramicrotomes including an Ultracut S, digital darkroom and wet darkroom facilities, along with several excellent LM with digital imaging systems.
Chair and Professor SUNY – BROCKPORT Department of Biological Sciences
The Department of Biological Sciences invites applications for a chair and professor. Foremost, the Department is seeking an energetic, visionary chair who will develop and enhance a balanced undergraduate major and Master’s degree program. Required: Ph.D. in biology or related field; strong teaching, scholarship, grant activity and administrative skills; ability to work collegially in culturally diverse environment. The disciplinary expertise of the candidates is a secondary consideration. In August 2001 the Department moved into a newly renovated building ($12 million) with state-of-the-art teaching and research facilities. Currently, the Department has strengths in environmental sciences, cell/molecular biology, and vertebrate biology.
Submit letter of application, resume, transcript showing highest earned degree, three letters of reference, and brief statements of leadership philosophy, teaching philosophy, research plans and equipment requirements.
Salary is competitive. Starting date is August 2002. Beginning review date for applications is December 15, 2001. Applications will be accepted until January 15, 2002 or until the position is filled.
Send application materials to: Mr. Terry Hooper, Faculty/Staff Recruitment Office, SUNY College at Brockport, 409 Allen Administration Building, 350 New Campus Drive, Brockport, NY 14420-2929 AA/EOE
Dr. Thomas P. Bonner Department of Biological Sciences SUNY at Brockport Brockport, NY 14420
We have been contacted by Agfa Corporation that they have a small inventory ( approx 30 boxes) of Agfa Scientia 23D56 Electron Microscope film. This film is 3.25" x 4" 100 sheet boxes, with an expiration date of 12/02. Agfa has said they will offer the film at a discount, but prefers to sell it in one bundle. If anyone is interested please contact me directly.
PS- If this is an improper use of this forum, I apologize. It seemed like the quickest method to distribute this information to those who may be interested.
Sincerely,
George
George Laing National Graphic Supply v:(518) 438-8411 X3109 v:(800) 223-7130 x3109 USA f:(800) 832-2205 email: scisales-at-ngscorp.com
We are about to retire a full size refrigerator which had been used to store osmium in both crystalline and liquid form for years. Our Safety people are asking us to clean/decontaminate it before they remove it from the lab. Is there any way to clean this ?
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
If anyone is interested, I have a Narishige Joystick Micromanipulator, model number MN151, for sale. I am asking $1300 or best offer. It has been used very little and it is still in the box.
If you are interested please e-mail me at j-kleier-at-northwestern.edu, off the list serve.
Morning Nami, While the dimensions are somewhat less than optimal, the following might offer two fruitful paths to try, and I emphasize might. I am reminded of an old, tried-and-true, AND direct approach which might help you quickly. Freeze drying of the unfixed organisms. Once the organism is dried, retrieval of the statolith should be facilitated via microdissection as long as it is not fractured by the freezing and drying procedure. While the above will probably enable isolation of the statolith, if I were anxious to study a statolith (with those dimensions) and in ascidian larvae, I would NOT try to isolate it, I would try to expose it in situ. The easiest method by which one might accomplish that is to deep freeze the entire structure, then cleave it with a single-edged razor blade, then look at it directly. If some of my preparations exposed an intact statolith, I would seek to isolate it after I had viewed it with the SEM and EDS/WDS. For TEM, I would process the structure in situ and thin section it as I would a piece of bone. Getting back to freezing and fracturing, with the proper equipment (VPSEM w/cleaving and cryotransfer attachment) you might be rewarded with much valuable data. In the absence of such equipment, I would concentrate first on the statolith which doesn't have to be preserved in the same manner as surrounding tissue. Now, how to accomplish this without expensive equipment. Since best fractures occur at lower temperatures, you might place a piece (brick) of steel (1" thick) in a small SS pan (NOT shallow) which is surrounded by the foam filler one can acquire at Home Depot as an insulator. Pre-cool the steel by pouring LN2 first over, then maintain it around the steel brick. Deep freeze the larva or the statolith, place it on the brick in a drop of an appropriate organic with a low freezing point to bind it to the brick, and cleave the frozen statolith with a pre-cooled single-edge razor blade held in a cooled pair of pliers held by your safely gloved hand. Tap the back of the razor lightly, though simply putting it down might be sufficient, to create a fracture, and hope for the best. NOTE: A member of my undergraduate class suffocated (and died) in an evaporated LN2 atmosphere. Please be careful how you construct an open LN2 system with which you must become somewhat intimate. Also, there are technical concerns beyond safety with which you should be familiar, so I recommend you 'study up' on the subject of freezing biological material for microscopic study before you proceed (if you decide to try this). You have asked for an approach, and I have suggested two. While the confocal might very well provide you with some interesting information about the hemisphere of surrounding tissue on the objective side of the organism, there is far less chance of acquiring any internal information since the statolith is not transparent and will thus be reflective no matter how you embed it. Good luck,
Fred Monson
} From:
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street
West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
To:
} Nami Choe } Ocean Science Centre } Memorial University of Newfoundland } St. John's, NF } A1C 5S7 Canada } phone:(709)737-3247 } fax:(709)737-3220 } } } } }
We sometime document our immunofluorescent images using regular SLR camera and Fujichrome 400 slide film. The problem is that the red fluorescent signal (568nm) did not show up bright on final developed film, However, the green (488) and blue (350nm) ones are fine. We use Olympus BX51 microscopy with a filter set of wide band (U-MWIY2)for red fluorescence.
Since the red signal is perfectly fine through binocular tube, I suspect that either the problem is caused by development (chemical) or film itself.
Any help will be highly appreciated.
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center 6000 Harry Hines Blvd., NA4.214 Dallas, TX 75390-9111
If there is black discolouration inside, it is due to insoluble osmium dioxide. You can possibly remove it by sponging with diluted hydrogen peroxide. This will reoxidise the dioxide to the tetroxide which is more soluble and can be washed off. Of course, in doing this, you will be exposed to osmium tetroxide solution and vapour, so you need protective clothing.
Otherwise you have to scrub off the dioxide with some kind of abrasive cream cleaner (do you have JIF?). The dioxide is inert and relatively harmless.
BTW adding hydrogen peroxide dropwise to osmium fixative which has gone dark will reoxidise it to the tetroxide (and water) and restore it to useful condition. Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
I feel somewhat out of parameters on this list; I have had some thoughts, however, about this topic.
I consider the work of the artist to open our eyes to new realities, to new ways of seeing ourselves, our experiences, the world around us. The artist is concerned to enhance our understanding of the world The physical product of the artist's work are not art, but artifacts. I think this definition of art is problematical, since there obviously is some sense of art work as an aethetic object as well.
Is the work of the scientist not also to shed new light that will enhance our understanding of the natural world?
Both are also concerned with sharing their work. Even the scientist who is cloistered in his corner of the laboratory must eventually be concerned to share new knowledge with others.
I wonder whether graphic data displays (as analytical tools) are objects of art? They can certainly be admirable.
The scientist is concerned with knowledge, with factual truth. This is a difference.
Another thought. I once read, though I don't remember where, a "Balinese saying." I cannot verify its authenticity:
We have no art. We do everything the best we can.
Is scientific writing a form of literary art?
Thank you for allowing me to express these disconnected thoughts.
Alan Davis Marianas High School Saipan, N. Mariana Islands adavis-at-saipan.com
I have just seen the new Nikon Super Coolscan 8000ED demonstrated. It was certainly very impressive. The 4000dpi and the density range of 4.2 make it a great scanner for EM negatives. I'm at present using a LeafScan 45 and this new Nikon was equal to or better than it in all areas and at a fraction of the cost. I have only a couple of reservations about it. The only negative holders that we saw demonstrated would not hold a standard (3 1/4 x 4 in , 8.8 x 10.2cm) TEM negative. We were forced to cut down the negatives. Has anyone been able to construct an adapter to take a full negative? I'm not too worried about the fact that it won't scan the entire area of the negative but don't like the idea of having to cut up negatives. It also seemed that the construction of the negative carriers did not appear to be very solid. I worry that in our multi user centre they may not last very long.
I'd be keen to hear from anyone who has experience with this scanner
Thanks
Rick --
=================================================== Rick Webb Senior Research Officer Department of Microbiology and Parasitology and Centre for Microscopy and Microanalysis University of Queensland 4072 Australia
Mel I purchased some ruthenium tetroxide about a year ago, but the stock went off quite quickly, and the ampoules are now full of a black precipitate. Can I titrate this against H2O2 in the same way ? Chris
} } BTW adding hydrogen peroxide dropwise to osmium fixative which has gone } dark will reoxidise it to the tetroxide (and water) and restore it to } useful condition. } Dr. Mel Dickson, } Deputy Director, The Electron Microscope Unit, } Adjunct Associate Professor, School of Microbiology & Immunology } The University of New South Wales } UNSW SYDNEY 2052 } Australia. } Phone +612 9385 6383 Fax +612 9385 6400 } http://srv.emunit.unsw.edu.au/ }
Most of us work with liquid nitrogen at some stage. Could you tell us more about the circumstances in which someone suffocated. I have up to now assumed small (1L) liquid volumes in a lab were safe. Last year a scientist died in Scotland. As far as I can tell a large volume of liquid was involved.
Dave
On Tue, 23 Oct 2001 12:18:35 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Morning Nami, } While the dimensions are somewhat less than optimal, the following } might offer two fruitful paths to try, and I emphasize might. } I am reminded of an old, tried-and-true, AND direct approach which } might help you quickly. Freeze drying of the unfixed organisms. Once the } organism is dried, retrieval of the statolith should be facilitated via } microdissection as long as it is not fractured by the freezing and drying } procedure. } While the above will probably enable isolation of the statolith, if } I were anxious to study a statolith (with those dimensions) and in ascidian } larvae, I would NOT try to isolate it, I would try to expose it in situ. } The easiest method by which one might accomplish that is to deep freeze the } entire structure, then cleave it with a single-edged razor blade, then look } at it directly. If some of my preparations exposed an intact statolith, I } would seek to isolate it after I had viewed it with the SEM and EDS/WDS. } For TEM, I would process the structure in situ and thin section it as I } would a piece of bone. } Getting back to freezing and fracturing, with the proper } equipment (VPSEM w/cleaving and cryotransfer attachment) you might be } rewarded with much valuable data. In the absence of such equipment, I would } concentrate first on the statolith which doesn't have to be preserved in the } same manner as surrounding tissue. } Now, how to accomplish this without expensive equipment. Since best } fractures occur at lower temperatures, you might place a piece (brick) of } steel (1" thick) in a small SS pan (NOT shallow) which is surrounded by the } foam filler one can acquire at Home Depot as an insulator. Pre-cool the } steel by pouring LN2 first over, then maintain it around the steel brick. } Deep freeze the larva or the statolith, place it on the brick in a drop of } an appropriate organic with a low freezing point to bind it to the brick, } and cleave the frozen statolith with a pre-cooled single-edge razor blade } held in a cooled pair of pliers held by your safely gloved hand. Tap the } back of the razor lightly, though simply putting it down might be } sufficient, to create a fracture, and hope for the best. NOTE: A member } of my undergraduate class suffocated (and died) in an evaporated LN2 } atmosphere. Please be careful how you construct an open LN2 system with } which you must become somewhat intimate. Also, there are technical concerns } beyond safety with which you should be familiar, so I recommend you 'study } up' on the subject of freezing biological material for microscopic study } before you proceed (if you decide to try this). } You have asked for an approach, and I have suggested two. While the } confocal might very well provide you with some interesting information about } the hemisphere of surrounding tissue on the objective side of the organism, } there is far less chance of acquiring any internal information since the } statolith is not transparent and will thus be reflective no matter how you } embed it. } Good luck, } } Fred Monson } } } From: } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } } To: } } } Nami Choe } } Ocean Science Centre } } Memorial University of Newfoundland } } St. John's, NF } } A1C 5S7 Canada } } phone:(709)737-3247 } } fax:(709)737-3220 } } } } } } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
LARGE VOLUME, SMALL SPACE - Truck cab in winter. Truck had a leak. Classmate closed windows of cab and simply went to sleep. Very sad and avoidable. I have found graduate students (still alive) in small closed office labs with open pans of LN2. They were unaware of the potential when air circulation was not assured.
Sorry for the unspecified reference.
Fred Monson
} ---------- } From: Patton, David } Sent: Wednesday, October 24, 2001 4:34 AM } To: Monson, Frederick C. } Cc: 'Microscopy Listserver'; 'Nami Choe' } Subject: Re: RE: microdissection before SEM/TEM } } Most of us work with liquid nitrogen at some stage. Could } you tell us more about the circumstances in which someone } suffocated. I have up to now assumed small (1L) liquid } volumes in a lab were safe. Last year a scientist died in } Scotland. As far as I can tell a large volume of liquid } was involved. } } Dave } } } On Tue, 23 Oct 2001 12:18:35 -0400 "Monson, Frederick C." } {fmonson-at-wcupa.edu} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Morning Nami, } } While the dimensions are somewhat less than optimal, the following } } might offer two fruitful paths to try, and I emphasize might. } } I am reminded of an old, tried-and-true, AND direct approach which } } might help you quickly. Freeze drying of the unfixed organisms. Once } the } } organism is dried, retrieval of the statolith should be facilitated via } } microdissection as long as it is not fractured by the freezing and } drying } } procedure. } } While the above will probably enable isolation of the statolith, if } } I were anxious to study a statolith (with those dimensions) and in } ascidian } } larvae, I would NOT try to isolate it, I would try to expose it in situ. } } The easiest method by which one might accomplish that is to deep freeze } the } } entire structure, then cleave it with a single-edged razor blade, then } look } } at it directly. If some of my preparations exposed an intact statolith, } I } } would seek to isolate it after I had viewed it with the SEM and EDS/WDS. } } For TEM, I would process the structure in situ and thin section it as I } } would a piece of bone. } } Getting back to freezing and fracturing, with the proper } } equipment (VPSEM w/cleaving and cryotransfer attachment) you might be } } rewarded with much valuable data. In the absence of such equipment, I } would } } concentrate first on the statolith which doesn't have to be preserved in } the } } same manner as surrounding tissue. } } Now, how to accomplish this without expensive equipment. Since best } } fractures occur at lower temperatures, you might place a piece (brick) } of } } steel (1" thick) in a small SS pan (NOT shallow) which is surrounded by } the } } foam filler one can acquire at Home Depot as an insulator. Pre-cool the } } steel by pouring LN2 first over, then maintain it around the steel } brick. } } Deep freeze the larva or the statolith, place it on the brick in a drop } of } } an appropriate organic with a low freezing point to bind it to the } brick, } } and cleave the frozen statolith with a pre-cooled single-edge razor } blade } } held in a cooled pair of pliers held by your safely gloved hand. Tap } the } } back of the razor lightly, though simply putting it down might be } } sufficient, to create a fracture, and hope for the best. NOTE: A } member } } of my undergraduate class suffocated (and died) in an evaporated LN2 } } atmosphere. Please be careful how you construct an open LN2 system with } } which you must become somewhat intimate. Also, there are technical } concerns } } beyond safety with which you should be familiar, so I recommend you } 'study } } up' on the subject of freezing biological material for microscopic study } } before you proceed (if you decide to try this). } } You have asked for an approach, and I have suggested two. While the } } confocal might very well provide you with some interesting information } about } } the hemisphere of surrounding tissue on the objective side of the } organism, } } there is far less chance of acquiring any internal information since the } } statolith is not transparent and will thus be reflective no matter how } you } } embed it. } } Good luck, } } } } Fred Monson } } } } } From: } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging(CASI) } } West Chester University of Pennsylvania } } Schmucker Science Center II } } South Church Street } } } } West Chester, PA, 19383 } } eMail: fmonson-at-wcupa.edu } } http://darwin.wcupa.edu/casi/ } } } } To: } } } } } Nami Choe } } } Ocean Science Centre } } } Memorial University of Newfoundland } } } St. John's, NF } } } A1C 5S7 Canada } } } phone:(709)737-3247 } } } fax:(709)737-3220 } } } } } } } } } } } } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
Listers, If anyone has available for sale a manual single or double-tilt specimen holder fitting the Philips CM 10/12 series, please contact me with a price quote and condition of holder. Thanks, Winston ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor 10/24/2001 10:29 AM Cannon Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 (Ship to: 1000 Blythe Blvd ) Fax: 704-355-0589 Charlotte, NC 28232-2861 (Ship to: 28203 ) Winston.Wiggins-at-CarolinasHealthCare.org {mailto:WWiggins-at-Carolinas.org} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ "Vocatus atque non vocatus, Deus aderit." - C.G.Jung
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I'm not sure exactly what info you're looking for, but we have adapters for the Coolpix to mount to just about any microscope. On the Nikon microscopes we have 3 options: a 23 mm phototube, a 30 mm phototube and a 38 mm ISO photoport. Give me a call and we can go over the specifics of your scope.
Tom Tottleben wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } To: Great Minds in the Halls of Science Wisdom! } } I realize this topic has been hammered out in this forum a few times } (for everything except the Nikon Optiphot superwide } trinocular) ......can someone direct me to any information that would be } helpful? ("I can do this .......yeah!!!") Thanks, } } Tom Tottleben } Tottleben Scientific Co } PO Box 24 } 104 Burns Farm-W.Ct. } Edwardsville, IL. 62025 } 618 656 9008 Office } 618 656 9599 Fax } 618 558 9008 Cell } 800 283 9997 Orders } tomtot-at-charter.net } www.tscmicroscopes.com
hello all! for those who work (or starting to work, that`s my case) on Bain tissue research: I`m looking for an rat/mouse antibody anti-NCAM and anti-PSA-NCAM. I recently purchased two antibodies from the Developmental Studies Hybridoma Bank of the U of Iowa, but those abs are anti-embrionic form of rat. I am looking for an `adult`rat/mouse form.
Please, I would appreciate your help
:)
Valeria L.Burgos Instituto de Ciencias Bàsicas y Medicina Experimental Hospital Italiano de Buenos Aires - ARGENTINA Tel: 54-11-9590200 extension 8919
_________________________________________________________ ¿Lo probaste? Correo gratis y para toda la vida en http://correo.yahoo.com.ar
I will appreciate to get any information on alignment of ion sources in Gatan DuoMill
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
I am wondering if there are folks who have used the Panasonic GP-US502 1/2" 3CCD camera and its successor the GP-US522 ?
We are thinking of integrating them on some microscopes.
Have you found GP-US502 sensitive enough to capture fluorescence images ?
Criticisms ? Prices Paid ? Thoughts ?
Thanks,
Sincerely,
Ed Monberg, General Manager LMDC 3101 Whipple Road, Union City, CA 94587 510-429-1060, Fax 429-1065, Cell 510-427-0115 WEB CATALOGUE: http://www.lasermotion.com ---------------------------------------------
To All who are concerned about my earlier LN2 reference.
In an earlier recommendation to the originator of the question about statoliths, I cautioned in my suggestions that a friend had died in an LN2 accident. Apparently there is some concern that I was referring to a death by a small amount of LN2. No one knows what volumes were involved, but the next paragraph should clarify the matter.
I didn't say that my friend died from a liter or, I think, suggest that the young lady could. My fraternity brother did die from asphyxiation from a nitrogen leak while he was trying to fix it in the closed cab of a tanker truck - according to the company AND the coroner at the time. We never could figure out why he was working in the cab or how there could have been a leak to fix in such a location. I even asked if it was the cab or the tank, and it WAS the cab. [A pressure line to the cab to monitor pressure in the tank? In their O2 carriers as well?????] All other info was lawyer'd quiet!
I never had a problem with a liter either, but I have recommended that students not work alone with any amount of LN2 in closed rooms with poor or no ventilation even if I couldn't specify the specific danger. I've never felt even drowsy either, but I haven't dealt with large-volume evolutions.
I did have an assistant once who loaded a 20L dewar in a small unventilated closet in which the 160L dewer was kept. When she came out, she claimed to feel faint. When I checked the dewar, it had lost half its contents in two days, and there appeared to be a leak in the siphon valve. I had it moved to a corner in a larger, well-ventilated room once the valve was replaced. Never had a problem thereafter, even when there were leaks.
In the final analysis, however, when I suggest the use of LN2 to people I don't know who work in environments with which I'm not familiar, I want a little "CYA" on my side.
Sorry I raised a red flag by an obscure reference to death. Perhaps it was ill-advised or at least poorly represented. I will not forget the concerns.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
Dear Listers I wish to sincerely thank everyone who responded to my question.This has opened more doors for me to explore.I am grateful for your generosity, in particular providing me with references and email contacts. Kind regards Judi Bowden
Roy, three easy ways to tell the difference between Si and GaAs:
1) GaAs is not as strong as Si. So GaAs wafers are generally thicker than Si wafers of the same diameter and are easier to break. 2) GaAs is a lot denser than Si. So if you have two wafers the same diameter but one is noticeably heavier (also because of (1)), it's GaAs. 3) GaAs cleaves on (110) and Si cleaves on (111). If your substrate has been cleaved up and has a flat edge perpendicular to the top surface, it's GaAs. If the edge orientation varies and is rarely perpendicular, it's Si.
Hope this helps!
Richard
} From: Beavers, Roy } Sent: Tuesday, October 23, 2001 4:44 PM } To: Microprobe List (E-mail) } Subject: Ni Films } } List, } } You have help me with issues before, which is greatly appreciated, so I } would like to ask help on the latest problem I have been working on. I } have } been looking at some 500 angstrom Ni films on Si substrates trying to } determine if there is any W contamination in the film. } } At first I thought I would use an beam of ~7KeV to keep my analysis within } the film as much as possible but I realized I would probably not see the } Ni } or W X-ray lines with this set up so I decided to go with a beam of 15KeV } which is where I usually run just to see what came up. } } Next I checked a Ni and W standard to see what lines I would get. This } looked o.k. so I proceeded to check a sample of the source material and as } expected I got a match for Ni and only Ni. } } Going to the film samples I expected to see Ni and I did, also I see no W, } but I did expect to see Si because of the higher beam energy but I did } not. } This is my first question why no Si lines? } } What I did see on the film samples that was unexpected was what I believed } to be lines for Ga and As. This may answer my question as to why no Si } signal but I want to check my thoughts on this before I tell my customer } they have deposited films on the wrong substrates. } } Any thoughts again will be greatly appreciated. } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu
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I'm looking for advice or anecdotes about moving heavy SEM equipment. I'm hoping to take possession of an old SEM, whose column base is 33"x33" and may weigh as much as 1,200 pounds. The console is wider and may weigh only 300 pounds.
If I could avoid $1,000 on professional movers and wrestle it with a few strong guys, that would be great, but there are other questions in my mind.
Regardless of who moves it, what considerations must they take regarding the sensitive nature of the equipment? Certainly sensitive bits like filaments should be removed and handled separately, right?
And what about the requirements for trucking? If it's going to spend two or three hours on the interstate, will any sort of truck do the job?
Hi All, To add to Fred's line about potential hazards involved with large volumes of LN2... Last spring, during the final stages of installing a new MRI facility here at the hospital a young man from the company doing the installation was killed by a large volume LN2 leak. I guess the coils of those beasts are cooled and there was a leak in one of the cooling lines and the room was filled with N2 gas...he was asphyxiated immediately. As long as our tanks & valves don't leak, the small volumes we actually deal with on a daily basis are probably not a problem, but I instruct everyone to work in a large room with good ventilation, or to leave the door open. I've had the ventilation in my EM room checked, since that is where my tank is stored.
lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
We are thinking to buy softwares which are good in both high resolution TEM image simulation and dynamical SAD and CBED diffraction simulation.
Can anyone recommend us some good ones?
Best Regards,
Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
I have a transformer for a Bausch and Lomb ring illuminator that I wish to donate to any public institution willing to pay the shipping charge ($ 5-10). I believe this works with the fluorescent ring illuminator for the stereozoom series. The electrical specifications are "PRIMARY: 115 V AC-60 cycles SECONDARY: 750 V 45 milliamps". It appears to be in good working condition (i.e. my voltmeter reads a 750 V output). I can send a digital image on request. Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Rick, Many of our customers are very happy with the 8000ED. I have looked into putting TEM negs into it and it could be done by modifying one of the 120/220 film holders. The holders have a raised lip to keep the film in the channel. I believe these could be removed and the film could then just extend out past the scan opening. I have not been able to try this as demand for the scanner has been very high. Another excellent scanner for TEM negs is the Agfa T2500 Duoscan. While lower in resolution(2500dpi optical) it has a glassless carrier design that will enable scanning of an entire TEM negative. It also will scan reflective originals.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
-----Original Message----- } From: Rick Webb [mailto:r.webb-at-mailbox.uq.edu.au] Sent: Wednesday, October 24, 2001 6:01 PM To: Microscopy Listserver
Hi
I have just seen the new Nikon Super Coolscan 8000ED demonstrated. It was certainly very impressive. The 4000dpi and the density range of 4.2 make it a great scanner for EM negatives. I'm at present using a LeafScan 45 and this new Nikon was equal to or better than it in all areas and at a fraction of the cost. I have only a couple of reservations about it. The only negative holders that we saw demonstrated would not hold a standard (3 1/4 x 4 in , 8.8 x 10.2cm) TEM negative. We were forced to cut down the negatives. Has anyone been able to construct an adapter to take a full negative? I'm not too worried about the fact that it won't scan the entire area of the negative but don't like the idea of having to cut up negatives. It also seemed that the construction of the negative carriers did not appear to be very solid. I worry that in our multi user centre they may not last very long.
I'd be keen to hear from anyone who has experience with this scanner
Thanks
Rick --
=================================================== Rick Webb Senior Research Officer Department of Microbiology and Parasitology and Centre for Microscopy and Microanalysis University of Queensland 4072 Australia
To All: I've been wondering whether to respond or not but here is a situation that happened in Vermont about 10 years ago. An artificial breeder was asphyxiated and died when he fell asleep in the pickup bed of his truck. He had his semen in a 5 - 10 gal LN2 Cryogenic Dewar that was in the bed of his pickup truck. I don't think he had a cab over the bed. The conditions were; warm/hot afternoon in Vermont (~80 degrees) cold for Texas. I believe the Breeder wanted to take a nap in the back truck bed. The Cold Nitrogen filled the truck bed and deplaced the Oxygen. The winds were minimal. He was overcome by a relative small amount of LN2 but there was a time factor. I believe they found him a few hours later. Beer may have also played a role.
Bottom Line: Don't sleep on the floor of the Lab if your using LN2
Dave Campbell IBM Micro-Electronics Essex Junction, Vt.
P.S. I was introduced to the Darwin Awards on this forum, but haven't seen any in a few years. Are there any updates? Please Respond off line.
I would not try to guess on this one. The only real solution is to have an oxygen monitor. They can be a pain because they are now required to be set at our facility to 19.5% for the alarm, so small spills can trip them. They also require weekly maintenance. I'd rather have the small annoyance of false alarms than the one time chance of being dead.
At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } To add to Fred's line about potential hazards involved with large } volumes of LN2... } Last spring, during the final stages of installing a new MRI } facility here at the hospital a young man from the company doing the } installation was killed by a large volume LN2 leak. I guess the } coils of those beasts are cooled and there was a leak in one of the } cooling lines and the room was filled with N2 gas...he was } asphyxiated immediately. } As long as our tanks & valves don't leak, the small volumes we } actually deal with on a daily basis are probably not a problem, but } I instruct everyone to work in a large room with good ventilation, } or to leave the door open. I've had the ventilation in my EM room } checked, since that is where my tank is stored. } } lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
Dear John, About two years ago I purchased a used SEM across the country and the movers only charged me about $500 to move it all that way. I chose a firm that delivers new SEMs, so they know what to do. You might ask your nearest SEM service provider for references. The microscope must be disassembled for shipping and the shipping bolts put in to stop things from shaking free. There may be some instructions with the documentation but at the least: 1. Remove the gun, cable and the connection into the high voltage tank and wrap up separately. Remove and box rotary pumps, hoses, power supplies and other connected pieces. 2. Label and disconnect all wires that stretch between the two sections. 3. Bolt down the column section. My moving company did not box the SEM, just put it on two pallets and wrapped it in plastic and moved everything, as much as possible, with a fork lift. Two strong men had no problem maneuvering the column section off the pallet and into the right place. If you think the column section is too heavy, you can probably take the lenses off, as well. This is a professional job to do right and IMHO the risk is worth getting an SEM service professional to pack it up and a scientific instrument moving company to move it is well worth what they charge. Make sure they have an "air-bed" truck for moving. At 07:23 AM 10/25/01 -0500, you wrote: } } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
I don't know anything about the stuctures you mention; however if you could work from a slice of tissue (e.g., for Xray), check out these publications:
Howell DN, Miller SE. Identification of viral infection by confocal microscopy. 1999. Methods in Enzymology. Academic Press, 307:573-91.
Miller SE, Levenson RM, Aldridge C, Hester S, Kenan DJ, Howell DN. 1997. Identification of focal viral infections by confocal microscopy for subsequent ultrastructural analysis. Ultrastruc Pathol 21:183-193.
Briefly, we make "vibratome" slices; stain them with propidium iodide, examine them by confocal microscopy for unusual areas, and then cut out the area of interest. Alternatively, if you have an antibody against the structure of interest, you can use a fluorochrome to label them. In t his manner, we've been able to locate a single virus-infected cell by confocal and then find that same cell by EM.
Good luck, Sara
On Mon, 22 Oct 2001, Nami Choe wrote:
} Date: Mon, 22 Oct 2001 15:37:43 -0230 (NDT) } From: Nami Choe {c56nc-at-morgan.ucs.mun.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: microdissection before SEM/TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear microscopy members; } } Hello. I am a marine biology graduate student at Memorial University of } Newfoundland, Canada. One of my research topic is to study the } microstructure of the statolith in the pelagic marine tunicate (Larvacea), } Oikopleura vanhoeffeni. The statolith is a small cylindrical calcium } build up (10 micrometer diameter, 20 micrometer height), attached to the } brain of the tunicate. This bony structure is contained in the } membraneous statocyst and probably works as a gravity detector. } I would like to isolate this structure for SEM and TEM analyses. After } this, Xray microcroprobe and other procedures will follow. } } Dissecting the brain is easy but taking out the statolith from the } statocyst is the difficult part. Visualizing the statolith can be done by } calcium specific stain, alizarin red. I have tried to dissect the } statolith out by using fine insect pins ( { 20 micron tip size), but the } pins are too large. Doing this under dissecting scope is impossible since } 4x magnification does not allow me to do such microdissection. } } I tried to dissolve the statocyst with bleach and hydrogen peroxide to } isolate inorganic statolith. However, after this procedure, I can no } longer visualize the statolith since alizarin red stain dissolves } instantaneously in these chemicals. } } Sonication was also done to rupture the statocyst and pop the statolith } out but it did not work. } } Do you have any suggestion on how I can separate this structure? } Sincerely, } Nami Choe } ____________________________________________________________________________ } } Nami Choe } Ocean Science Centre } Memorial University of Newfoundland } St. John's, NF } A1C 5S7 Canada } phone:(709)737-3247 } fax:(709)737-3220 } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Would the users of the SymParter attachment for the LKB knifebreaker care to comment on how useful it is? If you want to do it anonymously, e-mail me directly and I will post the replies without attribution. Thanks, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
on 10/25/01 5:23 AM, John Foust at jfoust-at-threedee.com wrote:
} I'm looking for advice or anecdotes about moving } heavy SEM equipment.
I've moved three SEMs, one of them locally, and two of them across-country towed behind (uncovered, but contents well-tarped) rental trailers . My experience has been that hiring SEM engineers is totally unnecessary.
our group works on Transmission Electron Microscopy of semiconducting materials, especially low-dimensional materials for optoelectronic applications. We work on InGaAs/GaAs single and stacked quantum dots and we recently started a new research activity on GaN on sapphire and/or Alumina substrates.
We have a good experience with TEM sample preparation, but we recently decided to buy the Multiprep System, to exploit new methods for TEM sample preparation and to minimize the ion milling time.
We just assembled and calibrated the machine, and we are now in the progress of preparing the first plan-view and cross-sectional samples.
As our machine is one of the few presently available in Italy, we are in contact with Allied to have suggestions and directions.
I should appreciate it very much if those of you who have operated the machine can give us suggestions, hints and procedures to get good quality samples with this nice machine.
Thanks in advance for your help
Massimo
Dr. Massimo Catalano IME-CNR Via Arnesano 73100 Lecce - ITALY phone: +39 0832 391199 fax: +39 0832 325299 email: massimo.catalano-at-ime.le.cnr.it
David this is how it should be done, but there is no point in using a monitor if it is disabled. A feature of the 1999 Scottish fatality was that the deceased habitually disabled the oxygen monitor. Having said that, we have operated without an oxygen monitor for years. Now may be a good time to buy one .... The circumstances of the Scottish fatality were somewhat unrepresentative of lab conditions in that having become unconscious, the victim was exposed to an effectively endless stream of nitrogen delivered from a large-volume dewar. My department responded to that incident by making it a rule that the person delivering nitrogen from a storage dewar should always be accompanied by a colleague. That way, there is a chance that someone can raise an alarm if things go pear-shaped.
Chris
} I would not try to guess on this one. The only real solution is to } have an oxygen monitor. They can be a pain because they are now } required to be set at our facility to 19.5% for the alarm, so small } spills can trip them. They also require weekly maintenance. I'd } rather have the small annoyance of false alarms than the one time } chance of being dead. } } } } At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould wrote: } } --------------------------------------------------------------------- --- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- --. } } } } } } Hi All, } } To add to Fred's line about potential hazards involved with large } } volumes of LN2... } } Last spring, during the final stages of installing a new MRI } } facility here at the hospital a young man from the company doing the } } installation was killed by a large volume LN2 leak. I guess the } } coils of those beasts are cooled and there was a leak in one of the } } cooling lines and the room was filled with N2 gas...he was } } asphyxiated immediately. } } As long as our tanks & valves don't leak, the small volumes we } } actually deal with on a daily basis are probably not a problem, but } } I instruct everyone to work in a large room with good ventilation, } } or to leave the door open. I've had the ventilation in my EM room } } checked, since that is where my tank is stored. } } } } lee } } -- } } Leona Cohen-Gould, M.S. } } Sr. Staff Associate } } Director, Electron Microscopy Core Facility } } Manager, Optical Microscopy Core Facility } } Joan & Sanford I. Weill Medical College } } of Cornell University } } voice (212)746-6146 } } fax (212)746-8175 } } } -- } David R. Hull } NASA Glenn Research Center at Lewis Field } Advanced Metallics Branch } Mail Stop 49-1 } 21000 Brookpark Road } Cleveland, OH 44135 } } (216) 433-3281 } fax (216)977- 7132 } david.r.hull-at-grc.nasa.gov } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html }
Even better, forced ventilation so that accumulation is impossible, and an alarm for when the impossible happens.
Ray } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } Organization: Inveresk Cottage } Reply-To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } Date: Fri, 26 Oct 2001 08:17:18 +0100 } To: "David R Hull" {David.R.Hull-at-grc.nasa.gov} } Cc: {microscopy-at-sparc5.microscopy.com} } Subject: Re: Potential LN2 Hazard? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } David } this is how it should be done, but there is no point in using a } monitor if it is disabled. A feature of the 1999 Scottish fatality was } that the deceased habitually disabled the oxygen monitor. Having said } that, we have operated without an oxygen monitor for years. Now may be } a good time to buy one .... } The circumstances of the Scottish fatality were somewhat } unrepresentative of lab conditions in that having become unconscious, } the victim was exposed to an effectively endless stream of nitrogen } delivered from a large-volume dewar. My department responded to that } incident by making it a rule that the person delivering nitrogen from } a storage dewar should always be accompanied by a colleague. That way, } there is a chance that someone can raise an alarm if things go } pear-shaped. } } Chris } } } I would not try to guess on this one. The only real solution is to } } have an oxygen monitor. They can be a pain because they are now } } required to be set at our facility to 19.5% for the alarm, so small } } spills can trip them. They also require weekly maintenance. I'd } } rather have the small annoyance of false alarms than the one time } } chance of being dead. } } } } } } } } At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould wrote: } } } } --------------------------------------------------------------------- } --- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } --------------------------------------------------------------------- } --. } } } } } } } } } Hi All, } } } To add to Fred's line about potential hazards involved with large } } } volumes of LN2... } } } Last spring, during the final stages of installing a new MRI } } } facility here at the hospital a young man from the company doing } the } } } installation was killed by a large volume LN2 leak. I guess the } } } coils of those beasts are cooled and there was a leak in one of the } } } cooling lines and the room was filled with N2 gas...he was } } } asphyxiated immediately. } } } As long as our tanks & valves don't leak, the small volumes we } } } actually deal with on a daily basis are probably not a problem, but } } } I instruct everyone to work in a large room with good ventilation, } } } or to leave the door open. I've had the ventilation in my EM room } } } checked, since that is where my tank is stored. } } } } } } lee } } } -- } } } Leona Cohen-Gould, M.S. } } } Sr. Staff Associate } } } Director, Electron Microscopy Core Facility } } } Manager, Optical Microscopy Core Facility } } } Joan & Sanford I. Weill Medical College } } } of Cornell University } } } voice (212)746-6146 } } } fax (212)746-8175 } } } } } } -- } } David R. Hull } } NASA Glenn Research Center at Lewis Field } } Advanced Metallics Branch } } Mail Stop 49-1 } } 21000 Brookpark Road } } Cleveland, OH 44135 } } } } (216) 433-3281 } } fax (216)977- 7132 } } david.r.hull-at-grc.nasa.gov } } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html } } } } }
I believe that in the case in Scotland the oxygen monitors were either switched off or ignored.
Dave
On Thu, 25 Oct 2001 17:03:58 -0500 David R Hull {David.R.Hull-at-grc.nasa.gov} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would not try to guess on this one. The only real solution is to } have an oxygen monitor. They can be a pain because they are now } required to be set at our facility to 19.5% for the alarm, so small } spills can trip them. They also require weekly maintenance. I'd } rather have the small annoyance of false alarms than the one time } chance of being dead. } } } } At 9:27 AM -0400 10/25/2001, Leona Cohen-Gould wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi All, } } To add to Fred's line about potential hazards involved with large } } volumes of LN2... } } Last spring, during the final stages of installing a new MRI } } facility here at the hospital a young man from the company doing the } } installation was killed by a large volume LN2 leak. I guess the } } coils of those beasts are cooled and there was a leak in one of the } } cooling lines and the room was filled with N2 gas...he was } } asphyxiated immediately. } } As long as our tanks & valves don't leak, the small volumes we } } actually deal with on a daily basis are probably not a problem, but } } I instruct everyone to work in a large room with good ventilation, } } or to leave the door open. I've had the ventilation in my EM room } } checked, since that is where my tank is stored. } } } } lee } } -- } } Leona Cohen-Gould, M.S. } } Sr. Staff Associate } } Director, Electron Microscopy Core Facility } } Manager, Optical Microscopy Core Facility } } Joan & Sanford I. Weill Medical College } } of Cornell University } } voice (212)746-6146 } } fax (212)746-8175 } } } -- } David R. Hull } NASA Glenn Research Center at Lewis Field } Advanced Metallics Branch } Mail Stop 49-1 } 21000 Brookpark Road } Cleveland, OH 44135 } } (216) 433-3281 } fax (216)977- 7132 } david.r.hull-at-grc.nasa.gov } http://www.lerc.nasa.gov/WWW/AdvMet/ASGWEB2000/asghome.html }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
John, You didn't state what kind of SEM. I've moved MANY ETECs with a 5x8 covered U-Haul trailer. I built a 10' folding ramp specifically for these low bed trailers and support the center with a couple of scissors jacks. I prefer to transport them under vacuum because the column is small enough to fit fully assembled and doesn't have a whole lot of mass. The optics table needs to be bolted down, as Mary said, and the high voltage supply handled separately, etc. The chief advantage on these is that they're on wheels and one person can do a move.
For systems that aren't on wheels (almost all others) you can rent a Johnson Bar (pry bar with wheels) and a safe jack (pair of 2 wheeled fork lift units that are strapped to the thing you want to move) and do it yourself (with a friend). You might want to consider a truck with a lift-gate and air suspension. Make sure things are well secured before you drive anywhere.
Good luck!
Ken Converse owner Quality Images third party SEM service Delta, PA
John Foust wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John } } } }
My experience is only anecdotal, but highly successful.
About a month ago I moved a JEOL-840 about 80 miles. The scope was donated to the University by a large corporation. It was decommissioned by a JEOL engineer, but it did not appear to have any special preparation other than to have the spring loaded column table bolted down. It was put on a pallet and moved to the shipping department by professional movers. From there on it was strictly an amateur job. I rented a Ryder truck, which I drove myself. Be sure the truck is high enough to do the job. I had to remove an ion pump at the top of the column in order to get the scope through the door of the truck. I unloaded it on my end with the help of one other person and a hydraulic pallet mover. There was a power lift available on both ends to match the height of the truck bed to the height of the loading dock. Two people (middle aged and not particularly hefty) were able to get the scope off the pallet and to its final position, using pry bars and 1/2 in aluminum rods as rollers. I did the installation myself, and it is now working much better than the similar but older scope it replaced.
If you have a few strong, but also smart helpers and some modest equipment, there is no need to fear the job. Work slowly and carefully. Feel free to call me if you want further details.
Dave Wilbur
-- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University voice: 617-627-2163 Fax: 617-627-3443 email: dwilbu01-at-tufts.edu __________________________________
----- Original Message ----- } From: "Paul B. Grover" {pbgrover-at-home.com} To: "John Foust" {jfoust-at-threedee.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 25, 2001 7:30 PM
Rick:
I second the notion that the AGFA T2500 Duoscan is and excellent scanner. My understanding is that the significant internals are made by Microtek which makes the AGFA T2500 nearly identical to the Microtek ArtixScan 2500 (another excellent scanner.) We also have customers quite pleased with these "twin" units. It's also worth noting, from what I've been told, that AGFA is discontinuing its T2500. I don't know if it's out of production yet, but I do know that because of the discontinuation, we've been able to offer good discounting to the customers we've proposed it to. If you choose to pursue the AGFA, (some inventory is still available) your dealer should be able to offer the same.
Good Luck.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045 *Microscopy and Digital Image Analysis for Science and Industry
-----Original Message----- } From: George Laing [mailto:scisales-at-ngscorp.com] Sent: Thursday, October 25, 2001 3:49 PM To: r.webb-at-mailbox.uq.edu.au; Microscopy-at-sparc5.microscopy.com
Rick, Many of our customers are very happy with the 8000ED. I have looked into putting TEM negs into it and it could be done by modifying one of the 120/220 film holders. The holders have a raised lip to keep the film in the channel. I believe these could be removed and the film could then just extend out past the scan opening. I have not been able to try this as demand for the scanner has been very high. Another excellent scanner for TEM negs is the Agfa T2500 Duoscan. While lower in resolution(2500dpi optical) it has a glassless carrier design that will enable scanning of an entire TEM negative. It also will scan reflective originals.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
-----Original Message----- } From: Rick Webb [mailto:r.webb-at-mailbox.uq.edu.au] Sent: Wednesday, October 24, 2001 6:01 PM To: Microscopy Listserver
Hi
I have just seen the new Nikon Super Coolscan 8000ED demonstrated. It was certainly very impressive. The 4000dpi and the density range of 4.2 make it a great scanner for EM negatives. I'm at present using a LeafScan 45 and this new Nikon was equal to or better than it in all areas and at a fraction of the cost. I have only a couple of reservations about it. The only negative holders that we saw demonstrated would not hold a standard (3 1/4 x 4 in , 8.8 x 10.2cm) TEM negative. We were forced to cut down the negatives. Has anyone been able to construct an adapter to take a full negative? I'm not too worried about the fact that it won't scan the entire area of the negative but don't like the idea of having to cut up negatives. It also seemed that the construction of the negative carriers did not appear to be very solid. I worry that in our multi user centre they may not last very long.
I'd be keen to hear from anyone who has experience with this scanner
Thanks
Rick --
=================================================== Rick Webb Senior Research Officer Department of Microbiology and Parasitology and Centre for Microscopy and Microanalysis University of Queensland 4072 Australia
We would appreciate any information/suggestions concerning service providers for a recently acquired Cambridge Stereoscan 240 SEM. To start we would be interested in a routine inspection and maintenance visit. We are located in Southern Connecticut. Thanks very much!
************************************* Christine Caragianis Broadbridge, Ph. D. Associate Professor of Physics Southern Connecticut State University 501 Crescent Street; Jennings 115 New Haven, CT 06515-1355
Hello list, It's been awhile since I needed work done on our Balzers 301 freeze fracture unit, so I'm asking who provides service on these instruments now? Randy Nessler University of Iowa Phone 319-335-8142
Can anyone give me their thoughts on the design of a request form for users of a microscopy core facility. I have done clinical TEM for twenty years but now I've been asked to manage a confocal/TEM core and will be getting a more diverse mix of specimens. What info should I request from the users? The obvious:1)project design- what questions do you wish to answer 2)sample type- cells, tissue etc. 3) fixed or live 4) fixation method 5) probes? 6) embedding method 7) etc. ?????
should there be two separate forms- one for TEM and one for Confocal? or one general form to include both microscopies?
What do you do? Maybe someone can fax me a copy of their labs request form that I can use as a template. Thank you all. Sincerely, Bill Oxberry Health Science Center Brooklyn fax # 718-270-3313
John: It can be done...I managed with the able assistance of a former student to shift a Hitachi 570 LB via a rental truck from the Smithsonian Institute in Washington, D.C. to Binghamton, NY. ` Success is dependent largely on the care taken with dismantling and packing. If possible, consult a service engineer or a firm that services SEMs of that manufacturer. They should be able to give advice on precautions to take prior to dismantling and shipment. Check out the SEM manual. It should give you exact dimensions and weights for each main part. The height can turn out to be all important...but more of that later. Plan on using lots of labeling tape to label connections. Stretch wrap which comes in 5" and 20" width rolls comes in useful. Boxed components should be plastic or stretch wrapped and well padded. There will be water hoses that will have to be disconnected and drained...and perhaps a water chiller to drain. Disconnect the vacuum hose and pack the rotary pump and make sure its kept upright. Disconnect connections between the display and column units. The cable connecting to the gun and high voltage tank should be removed and the ends stretch wrapped. Pack the cables with care to avoid bending too sharply. As with car spark plug leads the insulation could be damaged. Usually the column is suspended on anti-vibration pads. There should be bolts that can be applied to make the column firm for shipment. Ideally the column should shipped under vacuum. If you remove the top gun section, use a blanking plate to seal the rest of the column and rough pump before shipping. For strapping to pallets and to secure to truck interior you will find belt straps and plastic poly strapping come in handy. Strong backs help, but I would advise having the use of a "J" bar and a heavy duty pallet truck. And speaking of pallets... Just as we were on the loading dock and moving the column unit into the truck we discovered to our consternation we were just over one inch too tall to fit through the truck's door. So don't forget to figure in the height of the column PLUS the pallet when selecting a truck with a tall enough loading door. For us, this meant having to break the SEM column vacuum and remove the gun segment which housed the LB6 filament and ion pump, which fortunately was sealed off under it own vacuum. We covered the lower column with a metal plate and had to ship it without being under vacuum. Upon arrival back in Binghamton, NY the SEM had to be temporally stored in a hallway without access to a 208 V,30 AMP power supply. I had to rig up a system to keep the lower column under rough vacuum until the SEM could be properly installed in its permanent position. I had hole drilled into a half-inch thick aluminum plate cover and threaded to fit a pipe with a gas valve shut-off and a barbed end to fit a vacuum hose. This allowed me to rough-pump the column from the top. It was an interesting experience not to mention the coming and going to the Smithsonian Institute during a rather lively time when streets were blocked off during the World Trade Organization protests...but that's another story. Good luck with your move, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
John Foust wrote: } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John
-- ************************************************ Philosophy is the microscope of thought.
Victor Hugo, Book 3 Chapter 3 Les Miserables ************************************************ Laura Rhoads, Assistant Professor Biology Department SUNY Potsdam 44 Pierrepont Avenue Potsdam, NY 13676 315-267-2260 315-267-3170 fax
A postdoctoral position for electron holography studies of GaN structures is available for 14 months from 1 December 2001.
The project makes use of a Hitachi HF2000 FEGTEM equipped with an electron biprism and Gatan Imaging Filter and aims to develop methods of profiling electric fields which exist at interfaces and around defects. A good background in TEM is essential. Experience in electron holography is not essential.
For further details, contact Prof D. Cherns by e-mail, fax or phone as below
David Cherns H.H. Wills Physics Laboratory University of Bristol Tyndall Avenue Bristol BS8 1TL
Tel +44 (0)117 9288702 Fax +44 (0)117 9255624 E-mail D.Cherns-at-bris.ac.uk
If you want better semiconductor and GaN on sapphire samples than you can ever get by polishing and ion milling, may I suggest that you look into using the MicroCleave technique (also known as the small angle cleavage technique). I would recommend that you visit the South Bay Technology web site and check out the PDF files that they have on the kit. You will see an example of GaN on sapphire in one of their PDF files. There is no ion milling damage present at all. That was one of about five samples that was prepared in about two hours with the kit. Their web site is www.southbaytech.com.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it] Sent: Friday, October 26, 2001 2:53 AM To: Microscopy-at-sparc5.microscopy.com
Dear colleagues,
our group works on Transmission Electron Microscopy of semiconducting materials, especially low-dimensional materials for optoelectronic applications. We work on InGaAs/GaAs single and stacked quantum dots and we recently started a new research activity on GaN on sapphire and/or Alumina substrates.
We have a good experience with TEM sample preparation, but we recently decided to buy the Multiprep System, to exploit new methods for TEM sample preparation and to minimize the ion milling time.
We just assembled and calibrated the machine, and we are now in the progress of preparing the first plan-view and cross-sectional samples.
As our machine is one of the few presently available in Italy, we are in contact with Allied to have suggestions and directions.
I should appreciate it very much if those of you who have operated the machine can give us suggestions, hints and procedures to get good quality samples with this nice machine.
Thanks in advance for your help
Massimo
Dr. Massimo Catalano IME-CNR Via Arnesano 73100 Lecce - ITALY phone: +39 0832 391199 fax: +39 0832 325299 email: massimo.catalano-at-ime.le.cnr.it
} The Surface Science Laboratory at AECL Chalk River Labs has an opening for } a professional surface scientist. Anyone interested in applying is } encouraged to contact either Bill Hocking or Ian Muir. This position will } soon appear on the AECL website (www.aecl.ca) under Careers. Please use } this opportunity to apply on-line. } } Ian Bill } ---------------------------------------------- } ----------------------------------------------- } Ian J. Muir William H. Hocking, Ph.D. } Corrosion and Surface Science Branch Section Head, Surface Science } AECL, Chalk River Laboratories Corrosion and Surface Science Branch } Chalk River, Ontario AECL, Chalk River Laboratories } Canada K0J 1J0 Chalk River, Ontario, Canada } ph: (613) 584-8811 ext. 6960 K0J 1J0 } fax: (613) 584-3250 ph: (613) 584-8811 ext. 4651 } email: muiri-at-aecl.ca {mailto:muiri-at-aecl.ca} fax: (613) 584-3250 } email: hockingw-at-aecl.ca {mailto:hockingw-at-aecl.ca} } } } } Surface Science Laboratory } } The Surface Science Laboratory is part of the Corrosion and Surface } Science Branch within the Fuel Channels Division at AECL. The Laboratory } is equipped with a VG Scientific ESCALAB 220i-XL Imaging X-ray } photoelectron Spectrometer (XPS), a CAMECA ims 6F secondary ion } microanalyzer (SIMS), a Physical Electronics SAM 670 scanning Auger } microprobe and two JEOL scanning electron microscopes (SEM). An SEM/EDX } system is available for analysis of low activity samples, whereas highly } radioactive materials are accommodated in a shielded SEM/WDX facility. } The XPS and SIMS instruments are located in modern laboratories designed } for studies of highly radioactive samples and both systems have been } configured for safe handling of such samples. Activities in the Surface } Science Laboratory range from fundamental studies on nuclear materials to } commercial projects and failure analyses with a focus on characterization } of radioactive materials. } } Surface Science Position } } A research scientist is required to conduct studies of diverse materials } degradation and aging problems in the nuclear industry using secondary ion } mass spectrometry (SIMS) and other surface science methods. Applications } will be focused primarily on radioactive samples taken from reactor } components. } } Duties: } * Stress-corrosion cracking (SCC) of feeder pipes and steam-generator } tubes will be the principal area of initial focus, although support for } on-going studies of corrosion and deuterium ingress in pressure tubes will } be important as well. } * Development of expertise in other areas of applied surface science, } through participation in projects that require a multi-technique approach, } will also be expected. This will involve using scanning Auger microscopy } (SAM), imaging X-ray photoelectron spectroscopy (XPS) and scanning } electron microscopy (SEM) with energy dispersive X-ray (EDX) } microanalysis. } * Preparation of proposals for commercial and internal work, and } reporting to customers on such projects, will be key professional } activities. } * } * Qualifications: } * A graduate degree in surface science and a working knowledge of the } practical application of modern surface analytical methods, especially } SIMS, to industrial problems are required. } * Any additional training in, or experience with, electronics, } computers, spectroscopy, metallography, corrosion processes, radioactive } materials, or ultra-high vacuum technology would be an asset. } } The candidate must be a highly motivated individual with good written and } oral communications skills, who will be able to exploit the commercial } potential of the active surface science capabilities at CRL. }
Hi All, I would appreciate some practical help in choosing between the Leica (Reichert) Ultracut R and the Ultracut S on merit alone, notwithstanding considerations of price.
Thanks,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
The National Institute of Standards & Technology (NIST) has many Post Doctoral Positions open. These are offered competitively through the National Research Council (NRC). Within microscopy & microanalysis research areas we have many possible openings described at the web site listed below. The Microanalysis Research Groups at NIST have about twenty full time scientists specializing in the measurement methods in our extensive facilities including SEMs (FEG, EPMA, Environmental,…), AEMs (FEG/LaB6 TEM/STEM/EDS/EELS), Auger probes, SIMS/TOF-SIMS, MicroXRD/XRF, MicroRaman/IR, Synchrotron beam-line with grazing incidence XPS, etc.
We research new and improved microscopy and microanalysis measurement methods and we apply these methods to challenging analytical problems in semiconductor and optoelectronic technology, materials science, environmental and earth science, etc.
The NIST/NRC Post Doc program offers a two-year position at an annual salary of approximately $53,200 with an additional $5,500 for research expenses. The applications are due to the NRC by Jan 15, 2002. This must include a brief proposal and several recommendations.
A candidate must be a U.S. citizen and start work (with their PhD in hand) at NIST between July 2002 and January 2003. So, this is the perfect opportunity for those that are graduating this spring through next fall and others that have received their degree within the last five years. (Please note that NIST/NRC only competes Post Doc positions once a year, unlike some other institutions. )
Please contact me or go to the following site for further info:
We are in the process of ordering 30 student compound light microscopes for an upper level lab course. Our lowest bid ($400.00 below the next lowest) was for Wesco (or Para/Wesco) microscopes. Has anybody had any expierience with Wesco student scopes and can you make any recommendations. Thank you.
Mark J. Grimson
IMPORTANT NOTE: As required by a state of Texas mandate concerning formatting of state employee e-mail addresses, my new e-mail address is: mark.grimson-at-ttu.edu. The old one should continue to work for quite awhile, but you may want to go ahead and make this change in your address book. Thank you.
Mark J. Grimson, Ph.D Electron Microscopy Facility Dept. of Biology, MS 3131 Texas Tech University Lubbock, Texas 79409
One of the best student microscopes are for Meopta ( or Lambda) Czech rep. address is available on www.coleoptera.org in section scientific suppliers.
They have very good optical resolution, and very good design
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
----- Original Message ----- } From: "The Working Boy" {mark.grimson-at-TTU.EDU} To: "MSA" {microscopy-at-sparc5.microscopy.com} Sent: Saturday, October 27, 2001 7:37 AM
Dear all:
I know that somebody asked a question on how to observe paper structure. This is a little different case. I want to observe the structure of pressure sensitive film, Pressurex Film from Fuji. I am not aware enough of the film structure, but I wonder if I could see the ink capsules which normally breaks and provides red inks on being pressurized. If anybody could explain the film structure and/or the sample prep method, I would appreciate it. I am worried that the organic capsule would break and make mess under the e beam in SEM. If I make the film dry completely, would I see the capsule original spherical or like shrunk baloon?
Dear Jondo Yun, Many years ago I had a customer who routinely checked NCR paper for quality control, checking the size and distribution of the ink balls. I believe he just mounted and coated the samples. There was no problem with bursting spheres, etc.
Ken Converse owner Quality Images third party SEM service Delta, PA
Jondo Yun À±Á¸µµ wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all: } } I know that somebody asked a question on how to observe paper structure. This is a little different case. I want to observe the structure of pressure sensitive film, Pressurex Film from Fuji. I am not aware enough of the film structure, but I wonder if I could see the ink capsules which normally breaks and provides red inks on being pressurized. } If anybody could explain the film structure and/or the sample prep method, I would appreciate it. I am worried that the organic capsule would break and make mess under the e beam in SEM. If I make the film dry completely, would I see the capsule original spherical or like shrunk baloon? } } Best regards, } } Jondo Yun } jdyun-at-kyungnam.ac.kr } } }
I never used to have any problem, every so often I just sprayed with a multi-solvent household cleaner and the carbon coating on the inside of the bell-jar wiped off easily.
Now I'm using a different bell-jar, one that had been used for Al and that I had to clean up with acids, and I can't budge the carbon resulting from a few useages. I guess that the acid treatment somehow activated the glass surface so that the carbon really stuck.
Two questions:
How can I clean off this stubborn coating?
How can I prevent it from happening in the future?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Ritchie Sims wrote: ======================================================= I never used to have any problem, every so often I just sprayed with a multi-solvent household cleaner and the carbon coating on the inside of the bell-jar wiped off easily.
Now I'm using a different bell-jar, one that had been used for Al and that I had to clean up with acids, and I can't budge the carbon resulting from a few useages. I guess that the acid treatment somehow activated the glass surface so that the carbon really stuck.
Two questions:
How can I clean off this stubborn coating?
How can I prevent it from happening in the future? ======================================================== So far as removing what is already there, probably you are going to have to use some amount of good old-fashioned rubbing and scrubbing, using acetone and lint free cotton wipers. You can't do anything about the past but you can do something about the future.
And that is to use Bell Bright™ Bell Jar Spray, as shown on URL http://www.2spi.com/catalog/supp/supp3.html
A very thin layer of a water soluble polymer is left behind on the glass surface so that when it is time for a cleaning, a good lint-free cotton wiper and water alone, will cause the deposit to lift right off. In addition to saving a lot of hard work, you eliminate the breathing of acetone vapors.
Now for three words of further comment:
1] The formulation is not substantially different from a typical cosmetic hair spray, however it does not have the various additives that would otherwise gunk up a vacuum system.
2] It is a "gas under pressure" which means it is "dangerous goods" from an international shipment point of view, so it is expensive to ship by itself. It can be shipped only by air freight.
3] Disclaimer: SPI Supplies has formulated and sold this product since about 1978, so we have a vested interest in seeing more of it being consumed
I found your name on the website as someone who teaches existing server-side Java courses. My company has developed a new, practical server-side Java course, and we would like you to consider offering it to your students in the future.
You can view the syllabus of this new class that we developed at www.basebeans.com . The web site has the complete details of the Train the Trainer workshop. As technology advances from servlets to JSPs, the next step is towards JSP frameworks. As an instructor, I'm sure you are aware of the constant need to stay ahead of the curve.
If you would contact me at (415) 781-1463, I can tell you more. Thanks, Rene
1 Jiffy or another fine household abrasive. 2 Coat the inside of the belljar with dilute (roughly 1:20) dishwashing detergent. Dry before use. Less carbon will bounce off the coated belljar. So it gets dark faster, but it just about floats off with a bit of hot water. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, October 30, 2001 12:14 AM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi again } } I never used to have any problem, every so often I just sprayed with } a multi-solvent household cleaner and the carbon coating on the } inside of the bell-jar wiped off easily. } } Now I'm using a different bell-jar, one that had been used for Al and } that I had to clean up with acids, and I can't budge the carbon } resulting from a few useages. I guess that the acid treatment somehow } activated the glass surface so that the carbon really stuck. } } Two questions: } } How can I clean off this stubborn coating? } } How can I prevent it from happening in the future? } } } cheers } } rtch } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Version 4.0 of the Image Processing Tool Kit is now available. Free update copies have been sent to all registered purchasers of version 3.0, so if you neglected to send in the original registration card, now would be a good time to do it! New features, etc., are described at http://ReindeerGraphics.com
Am looking for a source to rebuild PGT detector. Crystal and/or the FET needs redone. I know PGT does things different than any of the other guys as far as the detector. Any names would be helpful. Thanks.
Joel McClintock EM Specialist U of Kentucky 859-257-1242
I once cleaned a very stubborn coating on a glass bell jar by using aluminum foil. Just crumple it up and start wiping. It takes time, but it will come off. I recommend that you use a product like bell bright on the glass prior to using the coater. It puts a layer between the glass and the deposit that is easily to take off with a wet cloth.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, October 29, 2001 9:14 AM To: Microscopy-at-sparc5.microscopy.com
Hi again
I never used to have any problem, every so often I just sprayed with a multi-solvent household cleaner and the carbon coating on the inside of the bell-jar wiped off easily.
Now I'm using a different bell-jar, one that had been used for Al and that I had to clean up with acids, and I can't budge the carbon resulting from a few useages. I guess that the acid treatment somehow activated the glass surface so that the carbon really stuck.
Two questions:
How can I clean off this stubborn coating?
How can I prevent it from happening in the future?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Technotrade International
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Mr. A. Auwärter (GM) / Mr. J. Hagen (Sales)
Mr. C. Tavernini ( Sales)
T: +1 603 622 50 11
F: +1 603 622 52 11
E-mail: albie-at-technotradeinc.com
E-mail: johnny-at-technotradeinc.com
E-mail: carlo-at-technotradeinc.com
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} Randy Nessler } University of Iowa } Phone 319-335-8142 } } }
} } Listers, } } Am looking for a source to rebuild PGT detector. Crystal and/or the } FET needs redone. I know PGT does things different than any of the } other guys as far as the detector. Any names would be helpful. } Thanks. }
Hi, Joel
I'm fairly sure that Gresham, of the UK, will rebuild any brand of detector. I think that Jim Nicolino, of X-Ray Optics, Florida, is their US representative.
But why not involve PGT?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I've been asked to look at some polystyrene microspherules in the SEM and would like some advice on how to prepare them. I'm not a biomedical person, so need some help!
The spherules are 5.6 um size with a Lumidin (protein) bonding surface and receptor molecules attached. They're in DI (or a DI solution?), which is apparently the delivery system for the marker molecules. Would they need CPD? If so, I've never CPD'd particles in solution before.... The SEM has hivac and VP modes, plus there's a cool stage.
Any ideas would be highly appreciated! Many thanks, Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
(Dr. Ritchie Sims on "Coating the Bell Jar" asked how to remove stubborn carbon deposits in a bell jar used to coat aluminum, then acid-etched, then reused with carbon.)
In making carbon fibers for composite materials, it is fairly common for manufacturers to activate the carbon surface (so that it will bond to matrix polymer or to a coupling agent) with an oxidizing-acid etch (often sulfuric or nitric). Dr. Sims, what acid(s) did you use, and at what concentration? Was the acid treatment applied again after carbon deposition, or only before? -------Roy
==================================== Roy Arrowood, Associate Professor Metallurgical and Materials Engineering UTEP, El Paso, TX 79968-0520 (915)747-6934 NEW E-MAIL ADDRESS: arrowood-at-utep.edu
I have been asked to demonstrate, by immunohistochemistry, the presence of sialodacryoadenitis virus, a rat coronavirus, in formalin fixed paraffin embedded tissue. Does anyone know of a commercially available antibody against SDAV that works in paraffin?
Unless they are microballoons, I wouldn't think you would need to do much special. I would probably start quick and dirty and get more complicated if necessary. I would try laying some down on a substrate and try crushing a few between glass slides to get a look at the interior to make sure the spheres are solid.
I would gold coat and use high vacuum mode in order to see much detail. I don't know what your clients want to know. We have some here trying to make small spheres and they are usually somewhat interested in the surface so we have to push for all the resolution we can get.
Warren
At 06:07 PM 10/29/01 -0500, you wrote:
} Hi listers, } } I've been asked to look at some polystyrene microspherules in the SEM and } would like some advice on how to prepare them. I'm not a biomedical } person, so need some help! } } The spherules are 5.6 um size with a Lumidin (protein) bonding surface and } receptor molecules attached. They're in DI (or a DI solution?), which is } apparently the delivery system for the marker molecules. Would they need } CPD? If so, I've never CPD'd particles in solution before.... The SEM has } hivac and VP modes, plus there's a cool stage. } } Any ideas would be highly appreciated! } Many thanks, } Dee } } *************************************************************** } Please do not publicly post any of my correspondence without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Peter: I've had to replace a number of parts recently for our Hitachi S 570 LB. All of the items were shipped promptly by Hitachi Instruments Inc. (800) 253-3053. In the future when parts may no longer be available from the manufacturer, I would try asking for help right here on the MSA list serve. Hopefully, someone will be able to come to the rescue. Henry
-----Original Message----- } From: Peter Tomic [mailto:PTomic-at-Anadigics.com] Sent: Monday, October 29, 2001 8:50 AM To: 'Henry Eichelberger'
John: It can be done...I managed with the able assistance of a former student to shift a Hitachi 570 LB via a rental truck from the Smithsonian Institute in Washington, D.C. to Binghamton, NY. ` Success is dependent largely on the care taken with dismantling and packing. If possible, consult a service engineer or a firm that services SEMs of that manufacturer. They should be able to give advice on precautions to take prior to dismantling and shipment. Check out the SEM manual. It should give you exact dimensions and weights for each main part. The height can turn out to be all important...but more of that later. Plan on using lots of labeling tape to label connections. Stretch wrap which comes in 5" and 20" width rolls comes in useful. Boxed components should be plastic or stretch wrapped and well padded. There will be water hoses that will have to be disconnected and drained...and perhaps a water chiller to drain. Disconnect the vacuum hose and pack the rotary pump and make sure its kept upright. Disconnect connections between the display and column units. The cable connecting to the gun and high voltage tank should be removed and the ends stretch wrapped. Pack the cables with care to avoid bending too sharply. As with car spark plug leads the insulation could be damaged. Usually the column is suspended on anti-vibration pads. There should be bolts that can be applied to make the column firm for shipment. Ideally the column should shipped under vacuum. If you remove the top gun section, use a blanking plate to seal the rest of the column and rough pump before shipping. For strapping to pallets and to secure to truck interior you will find belt straps and plastic poly strapping come in handy. Strong backs help, but I would advise having the use of a "J" bar and a heavy duty pallet truck. And speaking of pallets... Just as we were on the loading dock and moving the column unit into the truck we discovered to our consternation we were just over one inch too tall to fit through the truck's door. So don't forget to figure in the height of the column PLUS the pallet when selecting a truck with a tall enough loading door. For us, this meant having to break the SEM column vacuum and remove the gun segment which housed the LB6 filament and ion pump, which fortunately was sealed off under it own vacuum. We covered the lower column with a metal plate and had to ship it without being under vacuum. Upon arrival back in Binghamton, NY the SEM had to be temporally stored in a hallway without access to a 208 V,30 AMP power supply. I had to rig up a system to keep the lower column under rough vacuum until the SEM could be properly installed in its permanent position. I had hole drilled into a half-inch thick aluminum plate cover and threaded to fit a pipe with a gas valve shut-off and a barbed end to fit a vacuum hose. This allowed me to rough-pump the column from the top. It was an interesting experience not to mention the coming and going to the Smithsonian Institute during a rather lively time when streets were blocked off during the World Trade Organization protests...but that's another story. Good luck with your move, Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
John Foust wrote: } I'm looking for advice or anecdotes about moving } heavy SEM equipment. I'm hoping to take possession } of an old SEM, whose column base is 33"x33" and may } weigh as much as 1,200 pounds. The console is } wider and may weigh only 300 pounds. } } If I could avoid $1,000 on professional movers and } wrestle it with a few strong guys, that would be great, } but there are other questions in my mind. } } Regardless of who moves it, what considerations must } they take regarding the sensitive nature of the } equipment? Certainly sensitive bits like filaments } should be removed and handled separately, right? } } And what about the requirements for trucking? } If it's going to spend two or three hours on the } interstate, will any sort of truck do the job? } } - John
-- ************************************************ Philosophy is the microscope of thought.
Victor Hugo, Book 3 Chapter 3 Les Miserables ************************************************ Laura Rhoads, Assistant Professor Biology Department SUNY Potsdam 44 Pierrepont Avenue Potsdam, NY 13676 315-267-2260 315-267-3170 fax
I have an InP p-n junction device with Zn-diffused p-region. I would like to stain-etch the cross-section to observe the junction with SEM. Does anyone have a recipe for this stain-etch? Thanks.
Regards,
Judy Zhu, Ph.D. Member of Technical Staff Multiplex, Inc. 115 Corporate Blvd. South Plainfield, NJ 07080 phone: 908-757-8817 x 1167 fax: 908-757-8910 email: jzhu-at-multiplexinc.com
I've been attempting to do immunogold labelling of 4% paraformaldehyde fixed human natural killer cells (pre- and post- embedding). We're trying to label against perforin, but haven't had success. A monoclonal antibody we have labels the protein in cells fixed with methanol, but we need to preserve the ultrastructure of the cells. Is anyone using an antibody against the protein that works well in paraformaldehyde fixed cells? If so, would you please tell me what vendor you purchased it from, and perhaps share your protocol? Thank you.
Edward Haller University of South Florida Pathology Department Tampa, Florida(813)974-9584
I have become aware of a Community College student, Wil Kunkel, at Scottsdale Community College who will be joining this list in the near future. Wil takes some courses at AZ State U as well, and ended up on my door step with some questions re: SEMs.
It seems that SCC was the recipient of an ISI SX40 scanner from a local business which retired the instrument when its principal operator retired. Neither Wil nor the professor who was the receiver of the gift have any background in microscopy, but Dr. Sickafouse turned Wil loose on the scope after it was dumped in the room and somehow, Wil put it together and got it pumping. He next got a beam (with, as far as I can tell, no help) and started to make micrographs! Pretty good for someone with no EM experience, no vacuum system experience, and no help.
When we met, Wil was looking for information as to how he could get better micrographs. I was happy to spend a Saturday afternoon with Wil and Dr. Sickafouse and their merry band of budding microscopists, teaching some basic principles like gun alignment, signal maximization, vacuum cleanliness and protocols, etc. We were able to ascertain that the SEM was quite usable but will require a column liner dusting and cleaning so it can be stigmated and it will require another diffusion pump cleaning and replacement of the silicone based oil they used to begin with. (I hope they don't have too much trouble with contamination from that - it was a reasonable mistake for people with no Ebeam experience to make.)
My purpose here is to introduce Wil as a new microscopist who has no formal background (so his questions won't always be worded the way those of us who have some years in the business would do) but who has a remarkable interest, a willing mind, and a very high level of ability to reason and adapt - how many of us would - successfully- tackle the cleaning and replacement of an oil diffusion pump with no more help that a typical factory user's manual offers? How many of us could have buttoned up an old scope after a trip across town courtesy of amateur movers and got it to pump with no help?
To get to the purpose of this post - I can and have helped with generic questions and all-purpose adjustments, principles and practices, suggestions on writing a protocol so that new users can be trained and use the instrument for various research projects. What I can't do, and what I hope Wil and Dr. Sickafouse can find on the list, is help with their ISI SX40 specific questions and problems. Obviously, they don't have barrels of money and they have done very well already themselves with the nuts and bolts side of things, but there are sure to be questions that other ISI users and/or factory service people can help with much more expediently than can I.
Thanks in advance to all the generous listers who will, as always, I'm sure, help new people make it work.
Regards, Bill Sharp William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Here at the Central Michigan Biology Department Microscopy Facility we have found our lab not longer requiring the services of an old SEM that takes incredible images.. . . Lots of spare parts - contact me directly: willi1gl-at-cmich.edu
I am forwarding (with permission) a different take on electron microscopy service. This might be an excellent approach for private industry.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Excalibur Mineral Co [mailto:tony-at-excaliburmineral.com] Sent: Tuesday, October 30, 2001 8:27 AM To: tindallr-at-missouri.edu
We are still accepting applications for the position listed below
US-IL-Chicago Northwest-Electron Microscopist -- SEM
UOP LLC, located in northwest suburban Chicago, is an international leader in technology, products and services for the petroleum processing and separation industries. Our R & D Deptartment, which supports fundamental and market-focused R & D, has an immediate need for an Electron Microscopist to work with chemists and engineers throughout the company on structural and analytical characterization issues relating to catalytic, adsorbent, ion exchanger and metallurgy materials.
Technical assignments will include:
- Interaction and collaboration with solid state and inorganic chemists, applications specialists, and technical service personnel in applying appropriate microscopic and data analysis techniques to problem solving in catalytic and adsorbent materials fields. - Interaction and collaboration with existing microscopy group to effectively utilize SEM and EDXS facilities, including training of junior technical staff. - Development and continuation of in-house expertise in current and emerging scanning electron microscopy techniques and applications.
Minimum qualifications include:
- BS/MS with multi-year relevant work experience, or recent PhD in Geology, Chemistry, Materials Science, or Physics (preferred coursework in electron microscopy, solid state chemistry and physics). - Expertise in SEM and electron probe X-ray microanalysis techniques. - Knowledge of sample preparation techniques, including ultramicrotomy. - Excellent team and communication skills.
Knowledge of other microscopic characterization techniques, for example TEM and STEM, and industrial R&D experience desirable but not essential. Above all, we are seeking a motivated individual, who will demonstrate a willingness to learn and openness to acquiring new areas of expertise in characterization, chemistry and materials properties.
We offer a state-of-the-art scientific research environment coupled with exciting career opportunities coupled with a highly competitive compensation and benefits package including medical and dental coverage, 401(k) Plan, Pension Plan and tuition reimbursement.
US employment authorization required.
UOP is an Equal Opportunity Employer
Visit us at www.uop.com
---------------------------------------------------------------------------- ---- Additional Information Position Type: Full Time, Employee Ref Code: MB-01-11
---------------------------------------------------------------------------- ---- Contact Information UOP Internet Resume Manager cro-at-uop.com UOP LLC 25 E. Algonquin Road, P.O. Box 5017 Des Plaines IL 60017-5017 Fax: 847.391-2921
I have examined polystyrene beads under the SEM. It is not difficult to do. I applied a suspension of beads to a solid support with a pipetter, attached the support to an SEM stub, then sputter-coated the beads with gold. Once the sputter-coater pulls a vacuum, your solution will evaporate. I then viewed the beads under the SEM as usual, without any special attachments.
One thing you need to be careful of is losing the beads. Unless they are physically attached to your surface, they will not adhere to it. (Since they are spheres, they roll off, I guess.) If you have a steady hand this will not be a problem. The biggest challenge for me was smoothly inserting and removing the stub with the beaded support from our sputter coater. Several times I took the beads/stub over to another building to use the SEM. This involved walking down several flights of stairs, and I did not lose many beads (i.e., I saw numerous beads under the SEM.)
Hope this helps,
KD
-----Original Message----- } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] Sent: Monday, October 29, 2001 6:07 PM To: microscopy-at-sparc5.microscopy.com
A few colleagues and I have an interest in viewing SEM images of anthrax spores/virus. If someone can point me to a website that may have them in a library accessible to me, or if someone would forward some to me, we would greatly appreciate it.
Just so everyone is aware, this laboratory's principal function is the study of microelectronic failure mechanisms and not in any way related to biological technologies or biological warfare. We're simply curios.
Regards, Peter Tomic Anadigics, Inc. Warren, New Jersey
I am the lab coordinator here at ASUB. We have light microscopes with 100X oil immersion lenses. My supervisor wants me to attend a school of microscopy to enable me to better maintain and repair our microscopes. Do you know of available schools of microscopy that will meet my needs? Thanks, Linda
Rice University invites applications for a Staff Scientist position in the newly established Center for Biological and Environmental Nanotechnology (CBEN). Responsibilities include management of a major electron and probe microscopy facility, training of students in the use of the equipment, day-to-day maintenance of equipment, and participation in the research projects at Rice. The electron microscopy facility includes a JEOL 2010 TEM, JEOL 6320F field emission SEM, Phillips XL30 environmental SEM, Nanoscope AFMs, a Kamica Electron Microprobe and high-resolution XRD. This position requires a Ph.D. in Materials Science, Physics or a related field with specialization on electron microscopy. Preference will be given to the applicant with (1) experience in hands-on operation and maintenance of a range of electron and probe microscopes, (2) prior management experience, and (3) demonstrated research experience involving transmission electron microscopy. Interested persons should apply (include resume, statement of interest, list of publications, names and addresses of at least three references) by November 15, 2001 to:
Dr. Kevin Ausman Center for Biological and Environmental Nanotechnology MS-63 Rice University P.O. Box 1892 Houston, TX 77005-1892
*Rice University is an Affirmative Action/Equal Opportunity Employer*
I know this question was discussed in the past, but anyway. I would need a pretty comprehensive textbook(s)/ reference book(s) covering EM use in biology - possibly with stress on TEM, including sample preparation techniques. Any bright ideas?
Thanks,
Michael Jarnik
-- Michael Jarnik, Ph.D. Fox Chase Cancer Center Electron Microscope Facility Philadelphia
Semantics because this mistake has been making me crazy - ANTHRAX IS A BACTERIUM! This is a particularly important point because the word "virus" scares people silly. When I think of virus, I think of AIDS and Ebolla (sorry it that's misspelled). When I think of bacterium, I think antibiotic. No offense intended, Kristen
At 09:36 AM 31-10-2001 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011
Thanks to everyone for their helpful suggestions. Given below is the summary of all the replies I received regarding disposal of osmium contaminated refrigerator.
****************************
You can make them happy by moving the refer next to a hood and cleaning it with household bleach, the stronger the better. What this does is takes the relatively safe but dark osmium dioxide residue and converts it to the colorless but dangerous osmium tetroxide residue. It is generally impossible to decontaminate the surfaces, just cycle between toxic and nontoxic. The only other way is to rip out the contaminated areas of the interior of the refer and dispose of it in a toxic landfill. I'm sure they will appreciate this for milligram quantities of osmium. *****************************
If there is black discolouration inside, it is due to insoluble osmium dioxide. You can possibly remove it by sponging with diluted hydrogen peroxide. This will reoxidise the dioxide to the tetroxide which is more soluble and can be washed off. Of course, in doing this, you will be exposed to osmium tetroxide solution and vapour, so you need protective clothing. Otherwise you have to scrub off the dioxide with some kind of abrasive cream cleaner (do you have JIF?). The dioxide is inert and relatively harmless. BTW adding hydrogen peroxide dropwise to osmium fixative which has gone dark will reoxidise it to the tetroxide (and water) and restore it to useful condition. ***************************
There is often times a wide dichotomy between what safety people demand and what common logic and good sense suggest. If it is there at all, it would be black, right? Remember that if it was actually in the tetroxide form, it would be colorless and you would not see it, you would smell it, but actually it would disappear by itself because of its high vapor pressure. So if you have areas that are black, it would be the dioxide which is harmless. Just don't put in contact with any oxidizing agents which would reverse the reaction back to the tetroxide. You can clean it off however you think is the most convenient, perhaps with some laboratory Alconox, and then you could collect the reside removed and put it aside for recycling with your other precious metals slated for recyling and recovery. Unfortunately, too many "safety" people are starting to actual believe what they tell people like yourself. A few weeks ago, we had someone like yourself being told the refrigerator had to be sent to land fill and buried in its entirety! *********************************
100% ethanol will reduce the OsO4 to OsO2, and the latter is a harmless oxide.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
One of the particularly good ones is: Bozzola JJ, Russell LD, 1991. Electron Microscopy: Principles and Techniques for Biologist. Jones and Bartlett Publisher (Boston), 542 pp, ISBN 0-86720-126-6.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Wednesday, October 31, 2001 11:15 AM -0500 Michael Jarnik {M_Jarnik-at-fccc.edu} wrote:
} } Listers, } } I know this question was discussed in the past, but anyway. I would need } a pretty comprehensive textbook(s)/ reference book(s) covering EM use in } biology - possibly with stress on TEM, including sample preparation } techniques. Any bright ideas? } } Thanks, } } Michael Jarnik } } } -- } Michael Jarnik, Ph.D. } Fox Chase Cancer Center } Electron Microscope Facility } Philadelphia }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmichael-at-usc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, October 31, 2001 at 12:13:55 ---------------------------------------------------------------------------
Email: dmichael-at-usc.edu Name: Darren Michael
Organization: University of Southern California
Education: Graduate College
Location: Los Angeles, CA
Question: Hi,
I'm trying to find high index of refraction plastic coverslips. I've read about eyeglasses with high index of refraction (1.66) plastic lenses and have been unable to locate a manufacturer of coverslips using the same types of materials. Have you come across them? Do you have any suggestions as to whom to ask?
My department here is looking to acquire interactive scheduling software, preferably for the internet, that will cover all of our microscopes. Users should be able to create an account, manage it (add appointments, edit them, delete them...), and be able to see what time slots are available for the scope they wish to use. Please feel free to give me all suggestions that come to mind as to what software is available that will accomplish this. Thank you all very much.
Chris
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
Just wondering if anyone can give me some idea of the correct procedure for closing and opening both the column and gun valves when venting a Leica 440 SEM. Our manual only talks about closing the gun valve when using a Lab6 filament. The manual also talks about the gun valve 'flashing' when the vacuum is ready, and that at this point, the valve should be re-opened. Both the valves on our machine flash upon closing them and stay flashing until we return the leavers to the open position We are using a Tungsten Filment not a Lab6 and we do not have an Ion Pump on the column. We have the standard Rotary and Turbo pumps.
The manual does not explain when to close or open these valves when using a Tungsten Filament. My question is:- what is the correct procedure for closing the valves and re-opening them if we are using a Tungsten Filament and not a Lab6. At present we are closing the valves upon venting the machine in order to keep some vacuum in the column when the chamber is at air. As soon as we close the valves they flash red. Once we pump the column again, the gun and column valves stay flashing until we return the leavers to the open position.
Maxine Dawes Technical Officer School of Environmental Science and Management Southern Cross University PO Box 157 Lismore, NSW, 2480
I would like to do imaging of live GFP expressing mammalian cells. I am concerned about regulating pH without needing a gas flow system on the stage. Is adding HEPES to media sufficient to control pH in a bicarbonate based medium? Has anyone tried the Gibco CO2 independent medium to know if it is a high riboflavin medium (proprietary formula so I can't look it up). Thanks- Dave -- Permanent Address Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
Current Address 9/01-8/02 Dr. David Knecht School of Biosciences The University of Birmingham, Edgbaston, Birmingham, B15 2TT, U.K. Lab telephone: 0121 414 2508 (44-121 414 2508 from abroad) Department Fax: 0121 414 5925 (44-121 414 5925 from abroad) Direct Fax: 0121 414 5411 (44-121 414 5411 from abroad)
Try iCal {http://www.brownbearsw.com/index.html} , it may not be completely suitable for a large shared facility (somewhat limited use of alternate logins), but we use it on several smaller pieces of shared equipment (runs on NT). You can try the software as a shareware demo.
Another possibility is {http://www.loci.wisc.edu/calendar/} , which was described in a recent article in BioTechniques.
No commercial interest in either software. Doug
At 06:38 PM 10/31/2001 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good Evening all, } } My department here is looking to acquire interactive scheduling } software, preferably for the internet, that will cover all of our } microscopes. Users should be able to create an account, manage it } (add appointments, edit them, delete them...), and be able to see } what time slots are available for the scope they wish to use. } Please feel free to give me all suggestions that come to mind as to } what software is available that will accomplish this. } Thank you all very much. } } Chris
.................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (NEW email: Cromey-at-Arizona.edu) : :...................................................................: http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
I'm searching for freeze-substitution protocols (without using of any fixative), preferably fast and slow protocols, for plant leaf specimen preparation. I'm working on localizing nickel element in the plant leaves of a hyperacumulator plant, hence using cryo preparation methods. I'll highly appreciate any contributions and help in this.
My thanks in advance.
Chris Ogomo MSc Student University of the Western Cape Rep of S. Africa
Yeah, this book is really a good book for beginners. but there is now second edition in 1999 available 670pp, ISBN 0-7637-0192-0. The content is from basic structure and principles of TEM and SEM to sample preparation with many vivid pictures, mainly for tissue samples, also some pages on FT. This book is very basic, but not advanced, and it is quite cheap($8) .
Chen Chen
Department of Biological Chemistry The Johns Hopkins University School of Medicine 725 N. Wolfe Street Baltimore, Maryland 21218
On Wed, 31 Oct 2001 akc-at-umich.edu-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } One of the particularly good ones is: Bozzola JJ, Russell LD, 1991. } Electron Microscopy: Principles and Techniques for Biologist. Jones and } Bartlett Publisher (Boston), 542 pp, ISBN 0-86720-126-6. } } Kent } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } A. Kent Christensen, Professor Emeritus } Department of Cell and Developmental Biology, Medical Science II Building } University of Michigan Medical School, Ann Arbor, MI 48109-0616 } Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 } akc-at-umich.edu http://www.umich.edu/~akc/ } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } --On Wednesday, October 31, 2001 11:15 AM -0500 Michael Jarnik } {M_Jarnik-at-fccc.edu} wrote: } } } } } Listers, } } } } I know this question was discussed in the past, but anyway. I would need } } a pretty comprehensive textbook(s)/ reference book(s) covering EM use in } } biology - possibly with stress on TEM, including sample preparation } } techniques. Any bright ideas? } } } } Thanks, } } } } Michael Jarnik } } } } } } -- } } Michael Jarnik, Ph.D. } } Fox Chase Cancer Center } } Electron Microscope Facility } } Philadelphia } } } }
I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there.
One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there".
I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques.
My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them.
Any help you can provide would be very much appreciated - before I rip all of my hair out....
Thanks in advance, listers, to all of you. I know I came to the right place.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Electron Microscopy by John Bozzola and Lonnie Russell is excellent. I use it for both my TEM and SEM class. Biological Electron Microscopy by Michael J. Dykstra is also useful. It is more compact in size so easier om students' backs, and still seems to cover the necessary material. It doesn't include the extensive ultrastructure section Bozzola's book has. Either is good, and there are also many other good books out there. I have always spent a lot of time at the library display at the MSA meetings. Joyce Craig Chicago State University
Anthrax is actually the disease caused by the spore-forming bacterium Bacillus anthracis. The bacterium in question is actually named Bacillus anthracis. I guess it is easier for the news media to say anthrax than bacillus anthracis.
Tyrone Daulton
Kristen Lennon wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Semantics because this mistake has been making me crazy - ANTHRAX IS A } BACTERIUM! This is a particularly important point because the word "virus" } scares people silly. When I think of virus, I think of AIDS and Ebolla } (sorry it that's misspelled). When I think of bacterium, I think antibiotic. } No offense intended, } Kristen } } Kristen A. Lennon, Ph.D. } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } } 515-294-8854 } kalen-at-iastate.edu } www.baumlab.org
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Dear Collegues. I Have a Microlumina camera in my lab and lost the EasyScan 1.2 software as a result of a virus attack. The original diskas are by now unreadable. Does anyone have a copy of this software? I would be eternally gratefull if someone could send me a copy by e-mail to the address below. I think that the camera has been descontinued and is no longer available comercially.
Thanks in advance Dr. A.P. Alves de Matos Lisbon University Dental Medical School apmatos-at-ip.pt
Dear Dee, When I looked at latex spheres made in our Pathology department, I just dried a drop out of the suspension onto a polished graphite planchet. A glass cover slip is also a good substrate, if you are going to gold-coat anyway. If the carrier is really DI, it should dry away clean. If there is stuff in the DI, you will get smut around the spheres. I believe the polystyrene is strong enough to hold up to drying. If the spheres are small enough, they stick to the graphite themselves. If they are larger and in danger of rolling off, dry them onto a sticky tab. At 06:07 PM 10/29/01 -0500, you wrote: } } Hi listers, } } I've been asked to look at some polystyrene microspherules in the SEM and } would like some advice on how to prepare them. I'm not a biomedical } person, so need some help! } } The spherules are 5.6 um size with a Lumidin (protein) bonding surface and } receptor molecules attached. They're in DI (or a DI solution?), which is } apparently the delivery system for the marker molecules. Would they need } CPD? If so, I've never CPD'd particles in solution before.... The SEM has } hivac and VP modes, plus there's a cool stage. } } Any ideas would be highly appreciated! } Many thanks, } Dee Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hi Darren, Under separate cover I am sending an Excel table with plastics, their monomers and their indices of refraction. At least this may help you look.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: dmichael-at-usc.edu } Sent: Monday, October 22, 2001 5:13 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist: high index of refraction plastic } coverslips } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (dmichael-at-usc.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } October 31, 2001 at 12:13:55 } -------------------------------------------------------------------------- } - } } Email: dmichael-at-usc.edu } Name: Darren Michael } } Organization: University of Southern California } } Education: Graduate College } } Location: Los Angeles, CA } } Question: Hi, } } I'm trying to find high index of refraction plastic coverslips. } I've read about eyeglasses with high index of refraction (1.66) } plastic lenses and have been unable to locate a manufacturer of } coverslips using the same types of materials. Have you come across } them? Do you have any suggestions as to whom to ask? } } Thanks, } } Darren } } -------------------------------------------------------------------------- } - } }
Does anyone know of a most excellent method of fixing the mucin layers of a cell culture for TEM? There is a rumor of some osmium/fluoride compound that I'm trying to track down (osmium perfluouro-something?), but no luck yet. Personal experiences, references, and sources for exotic compounds would all be most welcome.
Thanks much.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
If you are teaching biological TEM, then you might consider using as a corollary "Cell and Tissue Ultrastructure: A Functional Perspective" by Cross and Mercer, pub by W.H.Freeman & Co NY. No sample prep or EM operation, but beautifully-presented micrographs illustrating ultrastructure and tissue types.
A.L.
####################################### Ann Hein Lehman Assistant Director, EM Facility Trinity College - LSC314 300 Summit Street Hartford CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman/
-----Original Message----- } From: Joyce Craig [mailto:J-Craig-at-csu.edu] Sent: Thursday, November 01, 2001 12:07 PM To: Microscopy microscopy
Electron Microscopy by John Bozzola and Lonnie Russell is excellent. I use it for both my TEM and SEM class. Biological Electron Microscopy by Michael J. Dykstra is also useful. It is more compact in size so easier om students' backs, and still seems to cover the necessary material. It doesn't include the extensive ultrastructure section Bozzola's book has. Either is good, and there are also many other good books out there. I have always spent a lot of time at the library display at the MSA meetings. Joyce Craig Chicago State University
At 09:21 AM 11/1/01 -0500, Chen Chen wrote: } Yeah, this book is really a good book for beginners. but there is now second } edition in 1999 available 670pp, ISBN 0-7637-0192-0. The content is from basic } structure and principles of TEM and SEM to sample preparation with many vivid } pictures, mainly for } tissue samples, also some pages on FT. This book is very basic, but not } advanced, } and it is quite cheap($8) .
Not sure about mucin, but have you looked into ruthenium red????
On Thu, 1 Nov 2001, Tindall, Randy D. wrote:
} Date: Thu, 1 Nov 2001 12:45:17 -0600 } From: Tindall, Randy D. {TindallR-at-missouri.edu} } To: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Subject: TEM: Mucin fixation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } Does anyone know of a most excellent method of fixing the mucin layers of a } cell culture for TEM? There is a rumor of some osmium/fluoride compound } that I'm trying to track down (osmium perfluouro-something?), but no luck } yet. Personal experiences, references, and sources for exotic compounds } would all be most welcome. } } Thanks much. } } Randy } } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
No one has mentioned this book, so I might as well: "Biomedical Electron Microscopy" by A.B. Maunsbach and B.A. Afzelius Academic Press, 1999, ISBN 0-12-480610-4 This text has many EMs comparing the effects of using different fixatives, buffers, embedding resins, and so forth. An excellent companion to Bozzola & Russell. Note: check ebay. I got my copy there for about 2/3 of the retail price.
Phil
} Electron Microscopy by John Bozzola and Lonnie Russell is excellent. I } use it for both my TEM and SEM class. } Biological Electron Microscopy by Michael J. Dykstra is also useful. It } is more compact in size so easier om students' backs, and still seems to } cover the necessary material. It doesn't include the extensive } ultrastructure section Bozzola's book has. } Either is good, and there are also many other good books out there. } I have always spent a lot of time at the library display at the MSA } meetings. } Joyce Craig } Chicago State University
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hello Paula, How many samples you need to examine depends on the details of the experiment, i.e. nature of specimens, the types of measurements taken, and most importantly the experimental question that is addressed by the experiments. The question to whether you have significant statistics is dictated by these enumerated, and perhaps other, factors. These factors will obviously differ for different experiments which have different goals.
My advice is to sit down and examine closely the experimental questions you choose to address and decide whether the data you have collected has the necessary statistics to adequately constrain the interpretations of the data.
The only meaningful reference (of which you seek) would be one that discusses measurement statistics on similar experiments, taking similar measurements, and addressing similar questions.
I hope this is of some help, good luck. Ty
Paula Allan-Wojtas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all, } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Even statistically speaking, the number of samples that you need depends on the variability between samples in your group and the extent to which the experimental group varies from the controls.
If your experimental group consistently looks one way and your controls consistently and obviously look different from the experimental group, I don't see why you would need to run statistics.
But it really depends on what you are trying to say. If all you are trying to point out is group one looks different than normal and it is obvious in all of your samples, (by obvious, I mean obvious to a trained eye) statistics is a waste of time. However, when you start to say group one looks somewhat worse than group two which looks somewhat worse than normal, and some of the specimens in the group actually look the same as, or better than ,the ones in the better group, then you'd need statistics. There are ways to figure out how big the group actually needs to be by taking a sample and determining the variability within and between groups (say on your 4 samples already obtained). I'm betting if you really can see something with these few samples, the estimation of sample size needed for statistics will be allot less than 30-40.
Unfortunately, I don't know of any articles you can site about the use of statistics in SEM off hand. Sorry. I've personally run into people recommending these ridiculously large sample sizes because they work with people. I work with animals and have allot more control over inter subject variability than they do, therefore I can work with a much smaller sample size even for statistics.
Good luck,
Karen Pawlowski, Ph.D.
Paula Allan-Wojtas wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all, } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
We regularly use 1% osmium tetroxide with 8% dimethoxypropane (DMP) in acetone for woody plant tissues with good success. The DMP removes water from the solution. We substitute for 5 days at -80 C. Anything less that 3 days won't work at all and more than 5 days makes no difference. We have used straight acetone for immuno work but the fixation (i.e. membranes) is not so great and the images are low in contrast. Other possibilities are 0.2 to 2% glutaraldehyde in acetone, with or without tannic acid. This is good for protein preservation for immuno work. Methanol is reported to work but not as well as acetone (I haven't successfully used it).
Let me know if I can be of further help.
Kim ------- Kim Rensing Ph.D. Dept. of Botany, UBC 6270 University Blvd Vancouver, BC, Canada V6T 1Z4
} I'm searching for freeze-substitution protocols (without using of any } fixative), preferably fast and slow protocols, for plant leaf specimen } preparation. I'm working on localizing nickel element in the plant leaves of } a hyperacumulator plant, hence using cryo preparation methods. I'll highly } appreciate any contributions and help in this. } } My thanks in advance. } } Chris Ogomo } MSc Student } University of the Western Cape } Rep of S. Africa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (griggsa-at-dbcc.cc.fl.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, November 1, 2001 at 14:26:59 ---------------------------------------------------------------------------
on 11/1/01 9:41 AM, Paula Allan-Wojtas at AllanWojtasP-at-EM.AGR.CA wrote:
} I am working on a multidisciplinary team of researchers on a number of very } interesting projects. I have a very large collection of top quality } micrographs from these projects which I would like to publish. The microscopy } samples have been selected and photographed within the experimental design } which has been set out for each project, so we are pretty sure they are } representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) } about my publishing any of the microscopy as part of any of the publications } put together by the team because we have not looked at 30-40 samples of each } of the many experimental treatments ( we have done 4 samples of each } treatment) . The other researchers on the team have looked at between 30 and } 40 samples per treatment. This one collaborator says that she has no } confidence in the microscopy results because "the statistics just aren't } there". } } I have tried on numberous occasions to explain to her about why this is not } feasible to do in SEM, and is usually not done. She does not believe me and } is essentially preventing me from publishing the work. I could probably } publish this work as a stand alone ms, but it is so much more meaningful when } integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why } 30-40 samples is not feasible for microscopy and which rationalizes why this } is so? The function of microscopy in the projects has been to give pictorial } information about the samples, and to suggest trends, not for absolute } measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of } my hair out.... } Dear Paula, As others have said, it depends on what kind of experiment you're looking at. Even if all the experimental samples look different from the controls, if you're trying to quantitate the changes--e.g., adding X to the culture shrinks the cells by Y%--you may still need sufficient data to make the numbers significant and to provide a standard deviation. Is is also important to have looked at different, independently-prepared samples, so if the few samples you observed were all from one experiment, I, too, would have doubts that they were sufficient. Then there's the problem that one selects what to photograph, and this selection always involves subjective factors. All that being said, however, if each of the other properties measured in the experiment were consistent from prep to prep, and if what you see correlates with what was measured, then, IMHO, it would be proper to publish the micrographs to illustrate what was found, regardless of how many or few you have. Good luck. Yours, Bill Tivol
Paula Most of us have grappled with this problem, but there is no generally applicable answer to the question "How many samples do I need to examine?". You are correct in saying that most electron microscopists don't attempt to examine 30-40 samples per treatment, but sometimes that is unavoidable. If the scientific question you are asking is a quantitative one - e.g. what proportion of cells have a particular morphology in normal and diseased tissues - and especially if the legal or public health or commercial cost implications of the answer are very significant - then it may be necessary to examine the number of samples and analyse the number of photographs demanded by the statisticians. If on the other hand you just want to illustrate the effect of a particular mutation on the shape of a fly's eye 2-3 representative samples are probably adequate. I feel hampered in making any really helpful suggestions about your experiment because it is not clear from your message what the nature of the experiment is, or what kind of observations you are attempting to make. Could you tell us more about that?
Best wishes Chris
} Paula Allan-Wojtas wrote: } } } -------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- ---. } } } } Hi, all, } } } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } } } Thanks in advance, listers, to all of you. I know I came to the right place. } } } } Paula. } } } } Paula Allan-Wojtas } } Research Scientist - Food Microstructure } } Agriculture and Agri-Food Canada } } Atlantic Food and Horticulture Research Centre } } Kentville, Nova Scotia Canada B4N 1J5 } } } } Tel: (902) 679-5566 } } FAX: (902) 679-2311 } } } } email: allanwojtasp-at-em.agr.ca
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Particles Are Tiny, but Damage Can Be Great
October 30, 2001
By JAMES GLANZ
Environmental scientists have spent decades studying the physics and physiology of particles very much like those in the most dangerous forms of biological weaponry.
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Thank you to everyone who has responded, so far. It has been very helpful for me. The responses have covered the whole spectrum from "yes, do the 30-40 samples" to "3-4 is probably enough". The majority of the answers fell in between these 2 points, saying that the number of samples required to be representative really depends on what you want/need to know from the samples and the sample variability. This could be partially achieved by taking pictures randomly over different samples. I will post a summary of answers soon. Anyone who does not want me to reveal their reply/name please contact me offline to let me know.
In some other unrelated work that we have done recently, we took the approach of working with a statistician from the outset. The project was a much more focused one, and we were looking for treatment effects, and intensity of these. One of the first steps was to actually do a small study to assess the variability of the population, and factors which affected it. With this information we were able to move to the next phase, actually testing the treatments. Statistics (experimental design, proper data collection and analysis of the data) allowed us to separate the population variability from the treatment effects, even when they were subtle. The result was information which did not agree with published literature or longheld assumptions which had arrived at through studies that did not have the "statistical rigour" that our studies did. Sorry to be vague about this, but it will be some months before I can openly talk about this work. I just wanted to say, for the record, that I am not against a statistical approach. I think it's a great idea, and I think it's the way to go.
In the original work I contacted the group about yesterday, we were just making some preliminary observations about the samples and suggesting that a connection could be made between the microscopy and other analytical methods. I have published other papers of this type with other workgroups, and everyone was ok with the "representative micrographs" idea, provided the usual disclaimers were used - "possible relationship" rather than "correlation"; "preliminary observations" ;More work should be done with more samples to be certain we're seeing what we think we're seeing " a first attempt at trying to relate microstructure to..."
I really can't go into more detail about this situation for obvious reasons. Suffice it to say that I plan to do my microscopy from now on in a new frame of mind, and will take some time to speak with a statistician before I start any new projects. My quest is to get the work I originally described, and all the work that we did connected to it, published in some form (complete with all the disclaimers necessary) as soon as I can.
Thanks again for all your help and support.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
We have been looking at marine algae in the same size range for quite some time. One of the considerations is having the sample distributed well enough to see individual particles and have a uniform random distribution for statistical purposes. Drying down from a drop of liquid tends to bunch up the particles and distribute the size ranges non-uniformly. We have used polycarbonate filters (Nuclepore, Poretics) and filter from liquid solution to collect the particles on a low profile background.
There are a number of pore sizes to choose from, select the one just below the size range of interest so that smaller detritus is removed. Try a dilution series to get a useable concentration. If the particles have enough inherent rigidity so they do not collapse, air drying is sufficient. We mount the filters on SEM stubs while still somewhat damp (they will fly off with static and/or handling of the flimsy material when completely dry). We then coat with gold. The one caution here is when the vacuum is applied and released make sure that it is done slowly so that large bursts of air don't sweep over the particles. Having said that our experience is that micron sized particles tend to stick to the surface they lay on. If you are adventurous you might try looking at the particles uncoated in VP mode.
Hope this helps.
-David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/micros_e.html
At 06:07 PM 10/29/01 -0500, you wrote: } } Hi listers, } } I've been asked to look at some polystyrene microspherules in the SEM and } would like some advice on how to prepare them. I'm not a biomedical } person, so need some help! } } The spherules are 5.6 um size with a Lumidin (protein) bonding surface and } receptor molecules attached. They're in DI (or a DI solution?), which is } apparently the delivery system for the marker molecules. Would they need } CPD? If so, I've never CPD'd particles in solution before.... The SEM has } hivac and VP modes, plus there's a cool stage. } } Any ideas would be highly appreciated! } Many thanks, } Dee
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } Does anyone know of a most excellent method of fixing the mucin layers of a } cell culture for TEM? There is a rumor of some osmium/fluoride compound } that I'm trying to track down (osmium perfluouro-something?), but no luck } yet. Personal experiences, references, and sources for exotic compounds } would all be most welcome. } } Thanks much. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
Ruthenium red, as Sara Miller wrote, works well. Osmium can be dissolved in organic compounds, the idea being not to expose the object of your study to aqueous solvents while it is being fixed. I don't know how well osmium fixes the mucin you are interested in. Glutaraldehyde and formaldehyde can also be "dissolved" or partitioned into organic solvents to accomplish the same end. This is known as phase partition fixation. I will send a reference if you like. Of course, if you are working with plastic culture dishes, the organic solvent must not react with the dish.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
} } } Dear Christopher, } } } The software that we use for scheduling is called Webevent. It is } } } fairly straightforward to use, and people can set up accounts and edit } } } them, depending on how you set it up. Their web site is: } } } http://www.webevent.com } } } Hope that helps. } } } } } } -Russell } } } } } } Christopher Gieczys wrote: } } } }
} } } } } } } } Good Evening all, } } } } } } } } My department here is looking to acquire interactive scheduling software, } } } } preferably for the internet, that will cover all of our } } } microscopes. Users } } } } should be able to create an account, manage it (add } } } appointments, edit them, } } } } delete them...), and be able to see what time slots are available for the } } } } scope they wish to use. Please feel free to give me all suggestions that } } } } come to mind as to what software is available that will accomplish this. } } } } Thank you all very much. } } } } } } } } Chris } } } } } } } } _________________________________________________________________ } } } } Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
-- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111
If you are not quantifying the microstructure with stereological measurements, then you use your qualitative judgment as to whether the micrographs that you use are representative. Typically you will get a feel for it. If on the other hand, you are using quantitative microscopy techniques, there are statistical tests that you judge your results by. These tests will tell you whether your sampling is sufficient. I recommend that you look at several books on the topic. The materials science oriented books that I am familiar with are DeHoff and Rhines "Quantitative Microscopy" and John Russ's books; try the "Image Processing Handbook" for example. John runs a short course on this topic every year and DeHoff is a co-instructor.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
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-----Original Message----- } From: Tyrone L. Daulton [mailto:tdaulton-at-nrlssc.navy.mil] Sent: Thursday, November 01, 2001 5:54 PM To: Paula Allan-Wojtas; microscopy-at-sparc5.microscopy.com
Hello Paula, How many samples you need to examine depends on the details of the experiment, i.e. nature of specimens, the types of measurements taken, and most importantly the experimental question that is addressed by the experiments. The question to whether you have significant statistics is dictated by these enumerated, and perhaps other, factors. These factors will obviously differ for different experiments which have different goals.
My advice is to sit down and examine closely the experimental questions you choose to address and decide whether the data you have collected has the necessary statistics to adequately constrain the interpretations of the data.
The only meaningful reference (of which you seek) would be one that discusses measurement statistics on similar experiments, taking similar measurements, and addressing similar questions.
I hope this is of some help, good luck. Ty
Paula Allan-Wojtas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, all, } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Or you could try Alcian blue/glutaraldehyde followed by osmium textroxide. See: Hear Research 1987;27(1):47-65 A newly identified surface coat on cochlear hair cells. Santi PA, Anderson CB.
Warning: This method is tricky because it can end up trapping water molecules in the mucous type tissues and it requires longer dehydration and polymerization times to set blocks hard enough for EM. But it looks great! (Actually, the molecules don't collapse as much in this as in ruthenium red, so the "coat" is closer to it's true thickness.) I only did some light level stuff with this myself, but I worked down the hall from Dr. Santi when this experiment was being done.
Karen Pawlowski
Sara Miller wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Not sure about mucin, but have you looked into ruthenium red???? } } On Thu, 1 Nov 2001, Tindall, Randy D. wrote: } } } Date: Thu, 1 Nov 2001 12:45:17 -0600 } } From: Tindall, Randy D. {TindallR-at-missouri.edu} } } To: "'microscopy-at-sparc5.microscopy.com'" } {microscopy-at-sparc5.microscopy.com} } } Subject: TEM: Mucin fixation } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } } } Does anyone know of a most excellent method of fixing the mucin layers of a } } cell culture for TEM? There is a rumor of some osmium/fluoride compound } } that I'm trying to track down (osmium perfluouro-something?), but no luck } } yet. Personal experiences, references, and sources for exotic compounds } } would all be most welcome. } } } } Thanks much. } } } } Randy } } } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.biotech.missouri.edu/emc/ } } } } } } } } } } } } } } Sara E. Miller, Ph. D. } P. O. Box 3712 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-3265
Dear Randy, Sorry for the bulk of this response, but finding what you needed required two Google searches, both of which might be of use to you along with the one finding that was close to what I expected. I recalled a paper in which SEM was performed on Ab-stabilized, then fixed, brush border. I couldn't remember that so I went searching - not for you, for me. I found: http://www.uth.tmc.edu/apstracts/1995/lung/December/226l.html. My two Google searches were: "glycocalyx fixation"; and "glycocalyx antibody"
Good luck,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I'm usually wary when someone is fixated on a particular "magic" number, 30 or 300 or whatever. Often you are dealing with a person who doesn't understand statistics nearly as well as they think they do. Since you haven't been able to convince her of your position, maybe you can ask her to justify hers? Make her really explain (and hopefully understand) where that 30-40 target comes from. I don't necessarily *recommend* this approach, since it may lead to more conflict than collaboration!
I don't know that a particular reference like the one you seek is available. As others have pointed out, it's hard to give specific advice without more specifics of your experiments. General suggestions might include 1) showing her other "comparable" microscopy examples from the literature; 2) explaining the total cost per sample in dollars and/or person-hours; 3) promising to include standard disclaimers or a brief discussion about the statistical relevance of the microscopy portion. I suspect you have already tried most or all of these. So lastly, about the only specific example I can think of is the various asbestos analysis regulations that attempt to apply statistics to microscopy (particle counting). There is a long history of trying to establish a minimum "representative" sub-sample based on sample heterogeneity. Feel free to email me if you think any of that may be helpful. Good luck!
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Thursday, November 01, 2001 9:41 AM, Paula Allan-Wojtas [SMTP:AllanWojtasP-at-EM.AGR.CA] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ------------------------------------------------------ ---- } -------------. } } } Hi, all, } } I am working on a multidisciplinary team of researchers } on a number of very interesting projects. I have a very } large collection of top quality micrographs from these } projects which I would like to publish. The microscopy } samples have been selected and photographed within the } experimental design which has been set out for each } project, so we are pretty sure they are representative of } what's there. } } One collaborator on the team is hesitant (to the point of } being resistant) about my publishing any of the } microscopy as part of any of the publications put } together by the team because we have not looked at 30-40 } samples of each of the many experimental treatments ( we } have done 4 samples of each treatment) . The other } researchers on the team have looked at between 30 and 40 } samples per treatment. This one collaborator says that } she has no confidence in the microscopy results because } "the statistics just aren't there". } } I have tried on numberous occasions to explain to her } about why this is not feasible to do in SEM, and is } usually not done. She does not believe me and is } essentially preventing me from publishing the work. I } could probably publish this work as a stand alone ms, but } it is so much more meaningful when integrated with data } from the other analytical techniques. } } My question: is there a reference (which I have somehow } missed) to explain why 30-40 samples is not feasible for } microscopy and which rationalizes why this is so? The } function of microscopy in the projects has been to give } pictorial information about the samples, and to suggest } trends, not for absolute measurement of structures and } changes in them. } } Any help you can provide would be very much appreciated - } before I rip all of my hair out.... } } Thanks in advance, listers, to all of you. I know I came } to the right place. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
} Email: griggsa-at-dbcc.cc.fl.us } Name: Alvin Griggs } } Education: Undergraduate College } } Location: Daytona Beach, FL, USA } } Question: I am looking for places that sell or lease microscope } projection oculars for use by visually impaired students. } } --------------------------------------------------------------------------- Alvin -
I would like to suggest that the current selection of add-on video cameras or direct digital input to a PC is a better way to go. You don't describe your application or budget, but simple video can be as cheap as $300. You'll find good descriptions of some possibilities at www.microscopeworld.com, but there are MANY sources.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
May 5 to 8, 2002, Palais des Congres de Montreal, Montreal, Quebec, Canada
held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
webaddress at the AOCS site: http://www.aocs.org/member/division/fsff/index.htm
Food Structure and Functionality Forum Bulletin Board: http://www.aocs.org/ubbcgi/ultimatebb.cgi
Tentative Technical Program Schedule (as of November 2nd, 2001)
Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl) --------------------------------------------------------------------------------------------------------------- Monday, May 6th-Morning Opening of symposium - Opening remarks
Plenary Speaker and presentation of Division Achievement Award
Dairy Applications Session. Chairs: Mark Auty, Dairy Products Research Centre, TEAGASC, Ireland (mauty-at-moorepark.teagasc.ie ) and Harjinder Singh, Massey University, NZ (H.Singh-at-massey.ac.nz)
Some Observations of a Microscopist, Author, and Reviewer on Structural Studies of Milk Products. M. Kalab, Agriculture and Agri-Food Canada, Canada
Effect of Emulsifiers and Processing Conditions on Microstructure of Milk Fat/Sunflower Oil Blends. S. Martini1, M. Cerdeira1, C. Puppo1, R.W. Hartel2, and M.L. Herrera1,3, 1CIDCA, UNLP, CONICET, Argentina; 1University of Wisconsin-Madison, USA; 3University of Buenos Aires, Argentina
Texturization of Dairy-Based Spreads. Y. Shi, B. Liang, and R.W. Hartel, University of Wisconsin-Madison, USA
Localization of Whey and Casein in Cheeses Using Microscopy and Immunochemistry Techniques. Y. Wang and D. Pechak, Kraft Foods, U.S.A
Manufacturing Yoghurt Structures with a Predicted Consumer Preference. M Langton and A. Astrom, SIK, Sweden
Dynamic Confocal Imaging of Tension and Fracture in Composite Food Materials. D.P Ferdinando1, K.P Plucknett2, and V. Normand3, 1Unilever Research, UK; 2DERA, UK; 3Firmenich SA, Switzerland
TBA - Topic: Dairy powders/caramels. C. Attapattu, University of Wisconsin, USA
Monday, May 6th - Afternoon Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl ) and David Pechak (Dpechak-at-kraft.com)
Protein Polysaccharide Interactions. C.G. De Kruif, NIZO Food Research, Netherlands (keynote speaker)
To Be Announced. B. Campbell, Kraft Foods, USA
Structure in Heat Treated Low_Fat Emulsions. R. Ofstad and V. Hoest, MATFORSK, Norway
Fatty Acid Salts-Induced Gels of Food Proteins: Their Rheological Properties and Structural Changes of the Proteins During Gelation. N. Yuno-Ohta, Nihon University, Japan
Wheat Gluten Proteins. A.S. Tatham, IACR Long Ashton research Station, United Kingdom
Interfacial Composition and Stability of Oil in Water Emulsion formed with Mixtures of Milk Proteins and Polysaccharides. H. Singh and Y. Hemar, Institute of Food, Nutrition and Human Health, Massey University, NZ
Dedicated Poster Session
Division Board Meeting -------------------------------------------------------------------------------------------------------------------- Tuesday, May 7th - Morning Agricultural Applications of Microscopy and Imaging Session/ joint with Feed Microscopy Division. Topic: Food Contamination
contacts: Mark Auty, Dairy Products Research Centre, TEAGASC (mauty-at-moorepark.teagasc.ie ) and Marge McCutcheon, West Virginia Department of Agriculture, USA (Feed Microscopy Division)
Forensic tampering. F. Platek, FDA (keynote speaker)
Contaminants in Food Processing. D. Kittleson, Pillsbury
How to approach contaminant identification. M. Auty, Dairy Products Research Centre
Growth promoters. P. Klink, South Africa
Identification of plant material. D.F. Wood, USDA
To Be Announced. J. Makowski
Quanitation of Total Fat and Fat Quality in Cheese and Dairy Products Using Membrane Separation Technology. V.C. Gordon, Safety Associates, Inc., USA
Additonal speakers to be announced.
Division Luncheon and round table (expert) discussion. Topic to be announced
Tuesday, May 7th - Afternoon Microbiology and Food Session Chairpersons: Judy Arnold and Ida Yates, USDA, ARS, Russell Research Center, USA
Prevention of Bacterial Fouling on Food Equipment Surfaces. J.W. Arnold, USDA, ARS, Russell Research Center, USA
Probiotics and Their Use in Food Animal Production. R. Droleskey, USDA, ARS, SPARC, USA
The Effect of High Pressure Steriliztion on Listeria Inoculated Seafood. K.R.S. Schneider and M.V.W. Wood, University of Florida, USA
Food Microstructure Investigations by Atomic Force Microscopy. J. Thornton, Digital Instruments/Veeco Metrology Group, USA
Controlling Growth of the Toxigenic Fungus, Fusarium Verticillioides. I. Yates and J. Arnold, Russell Research Center, ARS, USDA, USA
Division Members Meeting (immediately following the afternoon session) -------------------------------------------------------------------------------------------------------------------- Wednesday, May 8th- Morning Ingredients and Food Processing Session- Chairpersons: Diana Kittleson, Pillsbury Co, TPC Labs, USA; and Bernhard Tauscher, Federal Research Center for Nutrition, Germany
Non-Invasive Quality Determination of Fruit and Vegetables: Application of a Multi-Wavelength NIR-Diode Laser Array. B. Tauscher, Federal Research Center for Nutrition, Germany
Characterisation of the Application of Novel Oil Structurants. E. Floeter, F. Gandolfo, and W. Hogervorst, Unilever Research Vlaardingen, The Netherlands
High Pressure Processing. E. Ting and E. Raghubeer, Flow International, USA
Microstructure of Rice Starch Isolates. D.F. Wood1, A.M. Ibanez_Carranza2, and C.F. Shoemaker2, 1USDA, ARS, WRRC, USA; 2University of California, USA
To Be Announced. F. Escher, B. Conde-Petit, ETH, Switzerland
To Be Announced. M. Michel, Nestec Ltd., Nestle Research Center, Switzerland
To Be Announced. M. Salmenkallio-Marttila , VTT Biotechnology, Finland
Wednesday, May 8th - Afternoon New Methods and Techniques for Food Structure and Functionality Analysis Session Chairpersons: Kathy Groves, Leatherhead Food Research Association, England; and Maud Langton, SIK, Sweden
Quantifying Microstructures through Image Analysis. G.M.P. van Kempen1, M. van Ginkel2, C.L. Luengo Hendriks2, L.J. van Vliet2, and S. Singleton3, 1Unilever Research Vlaardingen, Netherlands; 2Delft University of Technology, The Netherlands; 3Unilever Research Colworth House, Great Britain
Cryo-TEM of Biopolymers in Comparision with Other TEM-techniques. M. Langton, A. Altskar, and A.-M Hermansson, SIK, Sweden
Freeze-substitution and low temperature embedding of dairy products for electron microscopy. A.K. Smith and H.D. Goff, Department of Food Science, University of Guelph, Canada
Recent Advances in our Understanding of the Relationship Between Crystallization Behavior, Microstructure and Rheological Properties of Fat Crystal Networks. A. Marangoni, University of Guelph, Canada
Spectroscopic Prediction of Rheological Properties in Grains. F. Meadows and F. Barton USDA, ARS, QARU, USA
Staining Techniques for Detection of Components in Fish Muscle. K. Hanneson Eggen and G. Enersen, Matforsk, Norway
Closure of Symposium ---------------------------------------------------------------------------------------------------------------------- Posters
Relationships between Microstructure and Rheological Properties of Model Lipid Systems. B. Liang, Y. Shi, and R.W., Hartel Univ. of Wisconsin-Madison, USA
Minor Biomolecules from the Olive Drupe to Olive Oil: The Technology and the Well-being Effects. N. Uccella, CIRASAIA-Mediterranean Agrifood Research Centre, Calabria University, Italy
Formation and Physical Properties of ?-Fat Gel. I: Macroscopic and Microscopic Observations. K. Higaki1, Y. Sasakura1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. Ii: In situ Observation of Gel-Formation Processes. K. Higaki1, Y. Sasakura1, I. Hachiya1, S. Ueno2, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. III: Rheological Properties. K. Higaki1, T. Koyano1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. IV: Why and How? K. Sato1, K. Higaki2, T. Koyano2, and I. Hachiya2, 1Faculty of Applied Biological Science, Hiroshima University, Japan; 2Meiji Seika Kaisha Ltd., Japan
} Date: Fri, 02 Nov 2001 07:40:47 -0800 } To: "Paula Allan-Wojtas" {AllanWojtasP-at-EM.AGR.CA} } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: SEM - how many samples are enough? } } Coming from a semiconductor background, I have a } somewhat different take on this topic. } } In evaluating the quality of IC wafer runs, there are } two methods I use. One way is to check each wafer } while the other is to check only one wafer. Either } way, only four die off of the wafer are examined via } SEM. This is in accordance with MIL-STD-883 Method } 2018, which can be found at: } } http://www.dscc.dla.mil/Downloads/MilSpec/Docs/MIL-STD-883/std883_2018.pdf } } In particular, look at paragraph 3.1.1 and Figure 2018-4. } } This area discusses die sampling procedures for wafer lots. } Considering that one wafer may contain upwards of 5,000 die } (depending on wafer size and die size), examination of } all die, or even a large number of die is unrealistic. The issue } is whether the sample is representative of the rest of the wafers } and the die therein. There are indeed strange things that can } and do happen to wafers as they are processed such that } the results of examining a randomly-pulled wafer may not } be correct about the whole wafer lot--or some of the other } wafers in the lot. } } If there is a reasonable expectation of repeatability from } wafer-to-wafer, then examination of one wafer and the } prescribed number of die will pass or fail the whole wafer } lot. perhaps this same reasoning might apply to your } situation? I certainly don't SEM 40,000 die! There are } in-process control wafers which are examined during } the course of fabrication. Failure at any of these steps } fails the whole wafer. But the small sample at the end } of fabrication is a pas/fail for the whole wafer lot. } } If you were to be examining E. coli for example and } imaged say ten individuals, and all were rod-shaped. } How many more specimens would you have to image } to find one that was round? The issue seems to me } to be more of the characteristics of the specimens } being examined. 100% coverage will always be } someone's desired approach. But of course, this } is not practical nor realistic for most situations. Thus, } the sampling criteria is applied. Which one is } applied is the key issue. How much or how many } is sufficient to draw a conclusion? } } gary g. } } } At 06:41 AM 11/1/2001, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We did a lot of freeze substitutions of plant leaf tissues, but with fixatives. However, I do believe the principle would be the same though without fixatives you would not clearly see the ultrastructure of the cells. Following are some references you might like to check.
Xu and Mendgen (1994) Planta 195:282 Xu and Mendgen (1997) MPMI 10:87
and some publications in Dr. K. Mendgen's lab.
Hope it helps.
Haixin Xu (Ph D)
Biological Sicences University od Maryland Baltimore County 1000 Hilltop Circle Baltimore, MD 21250
On Thu, 1 Nov 2001, Christopher Ogomo wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Greetings fellow listers. } } I'm searching for freeze-substitution protocols (without using of any } fixative), preferably fast and slow protocols, for plant leaf specimen } preparation. I'm working on localizing nickel element in the plant leaves of } a hyperacumulator plant, hence using cryo preparation methods. I'll highly } appreciate any contributions and help in this. } } My thanks in advance. } } Chris Ogomo } MSc Student } University of the Western Cape } Rep of S. Africa } } } } } } } }
Thanks to all the generous folks who sent me their microspherule protocols - The info is most appreciated! Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
I think this is a very interesting thread. I agree with most of the other comments contributed. It is the sort of problem that is not always clear but always important. In certain situations there are clear quantitative ways to arrive at the number of samples (measurements) required for a given confidence interval (key term), of say 95%, applied to a t-distribution assumption (usually). This type of formula can be plucked directly from statistical texts and there are even nifty little sample size calculators on the web. But, it is not always obvious how to apply that to a given measurement, and as others have said, we don't know exactly what you are trying to quantify. Within a given type of measurement, there are also ways to establish the precision of a given measurement method and then knowing that value, apply it to a single independent measurement acquired under conditions your accuracy/precision estimate was valid. There are very commonly scenarios where your final calculation also has errors that are cumulative and generally these add in quadrature. So, say you measure something and then calculate a result that involves ratios of this intensity or that, and/or multiplication of one length with another to get your final answer. Errors associated with division, multiplication and exponents are also known. In related schemes, say your calculation involves some intensity measurement and you know your the level of "noise" in that intensity measurement with a given RMS level, then that error may, in the most simple approximation, be given as 3X the rms and however that may be related to your final results would possibly be part of the quadrature-based calculation of your total error. I guess my point is there should be a way to estimate error within a given confidence interval in a single measurement if you know what the errors involved in the fundamental measurement, be it intensity, distance, etc. But, you need to break down that calculation and pick out the individual errors to determine the final error. Accuracy, versus precision, is a distinction I am sure you can appreciate.
I should say, I am definitely not a statistical expert but like you I have had to deal with this situation on several occaisons, as I am sure others have. Let's face it, it is also more work to generate error estimates for everything we do. Given the option we'd all like to plot and quantify with error bars and confidence limits, it is just not always clear how to "get there from here". My overall opinion is it is quite likely you have a valuable contribution and you need to determine some way to express qualitatively the limit in the interpretable information from the image (assuming it is an image) or quantitatively estimate that will satisfy your collegues. Good luck and please share any enlightment you might find as a results of this quest!
Regards, Ed Principe
} Paula Allan-Wojtas wrote: } } } -------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- ---. } } } } Hi, all, } } } } I am working on a multidisciplinary team of researchers on a number of very interesting projects. I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there. } } } } One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment) . The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". } } } } I have tried on numberous occasions to explain to her about why this is not feasible to do in SEM, and is usually not done. She does not believe me and is essentially preventing me from publishing the work. I could probably publish this work as a stand alone ms, but it is so much more meaningful when integrated with data from the other analytical techniques. } } } } My question: is there a reference (which I have somehow missed) to explain why 30-40 samples is not feasible for microscopy and which rationalizes why this is so? The function of microscopy in the projects has been to give pictorial information about the samples, and to suggest trends, not for absolute measurement of structures and changes in them. } } } } Any help you can provide would be very much appreciated - before I rip all of my hair out.... } } } } Thanks in advance, listers, to all of you. I know I came to the right place. } } } } Paula. } } } } Paula Allan-Wojtas } } Research Scientist - Food Microstructure } } Agriculture and Agri-Food Canada } } Atlantic Food and Horticulture Research Centre } } Kentville, Nova Scotia Canada B4N 1J5 } } } } Tel: (902) 679-5566 } } FAX: (902) 679-2311 } } } } email: allanwojtasp-at-em.agr.ca
Hi listers, I am looking for an ER specific fluorophore that can be excited using visible wavelength lasers. The cells that this would be used in are cultured fibroblasts. If possible, either the excitation or emission (one or the other, it needn't be both) would be different from those of Cy3 and Cy5. If anyone knows of any good kits available for this, I would greatly appreciate it (Mol. probes has a kit, but it requires UV excitation, a laser that I unfortunately do not have).
Thanks in advance, Russell -- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111 Tel. (617) 636-3795
I am forwarding the email from van Heel (sent to 3dem). It is a good summary of scheduling/booking softwares. I personally like iCal (http://www.brownbearsw.com/). Hope it'll help--Zhaojie Zhang
************************************** Zhaojie Zhang, Ph.D. Manager of Microscopy Facility University of Wyoming http://www.uwyo.edu/microscopy **************************************
-----Original Message----- } From: van Heel, Secretary T P M [mailto:vanheel.office-at-ic.ac.uk] Sent: Thursday, October 25, 2001 5:36 AM To: '3dem-at-sdsc.edu'
Paula writes ...
} ... I have a very large } collection of top quality micrographs from these projects which I } would like to publish. The microscopy samples have been selected } and photographed within the experimental design which has been } set out for each project, so we are pretty sure they are } representative of what's there. } } One collaborator on the team is hesitant (to the point of being } resistant) about my publishing any of the microscopy as part of } any of the publications put together by the team because we have } not looked at 30-40 samples of each of the many experimental } treatments ( we have done 4 samples of each treatment) . The } other researchers on the team have looked at between 30 and 40 } samples per treatment. This one collaborator says that she has no } confidence in the microscopy results because "the statistics just } aren't there". } ...
As others have implied ... how many samples depends on the variation you see. If you had 3 samples and absolutely no variation, you may be justified in stopping there and drawing a conclusion ... however, this is generally never the case. Statistical tools, such as the "chi-square distribution", are sometimes used for drawing conclusions from a small set, but even generalized chi-square formulas don't begin to yield meaningful conclusions unless the # of samples is ~30. This "rule-of-thumb" is probably at the root of your colleagues criticisms ... as if we remembered anything from "statistics 101".
The National Institute of Standards & Technology (NIST) and the Materials Science and Engineering Laboratory have post-doctoral positions open. These positions are offered competitively through the National Research Council (NRC). The Materials Structure and Characterization Group at NIST has many scientists utilizing our research facilities including TEMs (HRTEM, S/TEM, EELS, EDS) and SEMs (FEG, LaB6 equipped with OIM/EBSD).
We employ microscopy techniques to fundamental and challenging problems in materials science, semiconductors and optoelectronics. Recent topics have included semiconductor interconnections, nanometer scale multilayered materials, quantum dot materials, spintronic devices, etc.
The NIST/NRC Post Doc program offers a two-year position at an annual salary of approximately $53,200 with an additional $5,500 for research expenses. The applications are due to the NRC by Jan 15, 2002. The application must include a brief proposal and several recommendations.
A candidate must be a U.S. citizen and start work (with their PhD in hand) at NIST between July 2002 and January 2003. Students graduating this spring through next fall and others that have received their degree within the last five years are encouraged to apply. The NIST/NRC Post Doctoral program only selects positions once a year, unlike some other institutions.
You may contact me or visit our web site for further information at
Paula writes ... "I have a very large collection of top quality micrographs from these projects which I would like to publish. The microscopy samples have been selected and photographed within the experimental design which has been set out for each project, so we are pretty sure they are representative of what's there.
One collaborator on the team is hesitant (to the point of being resistant) about my publishing any of the microscopy as part of any of the publications put together by the team because we have not looked at 30-40 samples of each of the many experimental treatments ( we have done 4 samples of each treatment). The other researchers on the team have looked at between 30 and 40 samples per treatment. This one collaborator says that she has no confidence in the microscopy results because "the statistics just aren't there". "
} From Paul Webster: I am getting into this subject a little late so forgive me if I repeat something that has already been said.
The answer to the question, as posted by Michael S. a little earlier, is "... how many samples depends on the variation you see." This is exactly the solution to the problem. However, to get to a meaningful answer requires us to know what is the variation within the population. This may take a little more experimental planning. This point has been taken up in the rest of Michaels answer and also in previous replies concerning the examination of homogeneous materials samples. Homogeneous sample populations will require less sampling that heterogeneous populations.
To get a "representative" estimate of normal human blood temperature should be pretty easy as we all hover around the same temperature. Of course, a bias will be introduced if we include hospital patients in our sample. This is only a bias if we want an estimate for healthy individuals.
Conversly, obtaining an estimate for the mean height of a population will depend on who is being measured and their location. There will be many different, and correct answers for this one. Of course, the limitations of the estimation should be included with the final estimate (e.g. estimated mean height of adult males, age 35, attending a Yankees baseball game). Hwo many males should be measured so that we obtain a meaningful result? This will depend on the total population, their habits, and the sampling protocol applied. Selecting males going to the game who are seen coming out of "Big and Tall" stores may not be a representative sample!
Representative images in microscopy, selected to be "representative" are actually biased samples in that they have been selected by the operator. To get a truly "representative" result implies that unbiased estimators must be applied to the samples under investigation. An unbiased estimation should then be in the form of a quantitative result and not a "representative" image.
There are many powerful, efficient and easily applied protocols available for estimating number, length, surface area and volume. However, to be applied correctly, they must be applied to truly representative samples of the population. To obtain this unbiased sample requires the application of unbiased sample collection.
I saw in an earlier posting the cautious warning not to confuse accuracy with precision. This is an important warning. It is easy to be precise and still get the wrong answer. Imagine, for example, the result of a target practice using a gun with crooked sights but in the hands of an exceptional marksman. The holes will probably all be clustered together. The bias due to the crooked sight may cause the holes to be way of center. However, in the hands of a novice, the gun will fire off-target, but the operator may possibly be so inaccurate that a stray shot will hit the bulls-eye.
In the words of the sterologists "do more - less well". In biology, most of the variation occurs at the level of the cell or the animal. There will be less variation between samples taken from the same animal or cell population. It is far more reliable to measure small samples from more animals or cells than to take lots of images from one sample.
There are many unbiased sampling protocols available to use. However, most of them are designed to be applied to thin sectioning. It is possible that they can be adapted for use in the SEM. Estimating number and particle size should be possible using ramdom fields.
To obtain more information on these protocols I suggest looking up papers by John Lucocq and Terry Mayhew, who have both written very readable reviews of the challenges of random sampling. In particular, look for descriptions of Systematic Random Sampling. You will also see references to uniform isotropic sampling to eliminate bias.
You may come to the conclusion that taking one or two random images from your 30-40 samples, and then estimating the paramenters of interest from them is the way to go.
Microscopy has become a powerfully quantitiative branch of science. As such it is possible to express the qualitative results we see every day in unambiguous ways. We should make use of these tools to remove the fuel for argument.
In setting up the monthly archives for October of 2001 I got moderatley curious about the number of Email messages/ deliveries have run through the system running out of my home/office these past 8 years (Oct 1, 1993 was the Listserver's start date).
According to my extremely precise calculations ;-) the 100 Millionth Email message was delivered to a Microscopy Listserver subscriber sometime this year.
In case your curious, there have been 58, 280 postings to the Microscopy Listserver over the last 8 years, all of which are stored in the searchable archives. The subscriber base varies (daily), but as of this morning we had 2562 subscribers from at least 62 countries on 6 Continents. There is no one from Antarctica that I can find, or shall we say that I know about, nor is anyone from the International Space Station subscribed. :-( We did have at least one person subscribed whose mail is being forwarded to an Oceanographic survey vessel via a satellite link.
Not counted in the above totals were 6415 "suspect" messages caught by the Email filter and were rejected. Just under 3000 of these had attachments, 700+ had forged IP addresses, another ~2000ish were offers for credit/mortages/loans/viagra/porn etc... but at the same time number of valid postings { 3 % were inadvertently caught and these are generally passed through after I get contacted by the authors asking what is wrong.
I also noticed that the very first subscriber to the Listserver (Arthur Day from ANSTO just outside of Sydney Australia) is still receiving his daily dose of Microscopy Email. Well done, Arthur!
Now if I only had a beer for each of those emails, I'd be set for life.
} } Now if I only had a beer for each of those emails, I'd be set for life. { {
Actually Nestor, by my extremely precise calculations, someone should buy you your 100,000,000th beer about MSA time next year...something to look forward to in Quebec City!
Larry :-)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listserverites... } } In setting up the monthly archives for October of 2001 } I got moderatley curious about the number of Email messages/ deliveries } have run through the system running out of my home/office } these past 8 years (Oct 1, 1993 was the Listserver's start date). } } According to my extremely precise calculations ;-) the 100 Millionth Email } message was delivered to a Microscopy Listserver subscriber } sometime this year. } } In case your curious, there have been 58, 280 postings to the Microscopy } Listserver over the last 8 years, all of which are stored in the } searchable archives. The subscriber base varies (daily), but as of this } morning we had 2562 subscribers from at least 62 countries on 6 } Continents. There } is no one from Antarctica that I can find, or shall we say that I know about, } nor is anyone from the International Space Station subscribed. :-( } We did have at least one person subscribed whose mail is being } forwarded to an Oceanographic survey vessel via a satellite link. } } Not counted in the above totals were 6415 "suspect" messages caught } by the Email filter } and were rejected. Just under 3000 of these had attachments, 700+ } had forged IP addresses, another ~2000ish were offers for } credit/mortages/loans/viagra/porn etc... but at the same time } number of valid postings { 3 % were inadvertently caught and these are } generally passed through after I get contacted by the authors asking } what is wrong. } } I also noticed that the very first subscriber to the Listserver } (Arthur Day from ANSTO just outside of Sydney Australia) } is still receiving his daily dose of Microscopy Email. Well done, Arthur! } } Now if I only had a beer for each of those emails, I'd be set for life. } } Cheers... } } Nestor } Your Friendly Neighborhood SysOp
-- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
for symmetry I think Arthur Day should be the one to buy Nestor his 100,000,000th beer, and the 17th Australian Conference on Electron Microscopy in Adelaide this coming February would be the perfect place. Adelaide is quite warm that time of year , perfect beer drinking conditions. Why don't all you listservers come and join us. Cheers,
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} ---------- } From: Larry Allard } Sent: Monday, November 5, 2001 7:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Administrivia: 100,000,000 and counting } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Nestor states: } } } } Now if I only had a beer for each of those emails, I'd be set for } life. { { } } Actually Nestor, by my extremely precise calculations, someone should } buy you your 100,000,000th beer about MSA time next year...something } to look forward to in Quebec City! } } Larry :-) } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Listserverites... } } } } In setting up the monthly archives for October of 2001 } } I got moderatley curious about the number of Email messages/ } deliveries } } have run through the system running out of my home/office } } these past 8 years (Oct 1, 1993 was the Listserver's start date). } } } } According to my extremely precise calculations ;-) the 100 Millionth } Email } } message was delivered to a Microscopy Listserver subscriber } } sometime this year. } } } } In case your curious, there have been 58, 280 postings to the } Microscopy } } Listserver over the last 8 years, all of which are stored in the } } searchable archives. The subscriber base varies (daily), but as of this } } morning we had 2562 subscribers from at least 62 countries on 6 } } Continents. There } } is no one from Antarctica that I can find, or shall we say that I know } about, } } nor is anyone from the International Space Station subscribed. :-( } } We did have at least one person subscribed whose mail is being } } forwarded to an Oceanographic survey vessel via a satellite link. } } } } Not counted in the above totals were 6415 "suspect" messages caught } } by the Email filter } } and were rejected. Just under 3000 of these had attachments, 700+ } } had forged IP addresses, another ~2000ish were offers for } } credit/mortages/loans/viagra/porn etc... but at the same time } } number of valid postings { 3 % were inadvertently caught and these are } } generally passed through after I get contacted by the authors asking } } what is wrong. } } } } I also noticed that the very first subscriber to the Listserver } } (Arthur Day from ANSTO just outside of Sydney Australia) } } is still receiving his daily dose of Microscopy Email. Well done, } Arthur! } } } } Now if I only had a beer for each of those emails, I'd be set for life. } } } } Cheers... } } } } Nestor } } Your Friendly Neighborhood SysOp } } -- } Dr. Lawrence F. Allard } Distinguished Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 865-574-4981 } 865-576-5413 Fax } allardlfjr-at-ornl.gov } }
Does anybody have the information about the using possibility of image recording systems (like PIXsysTEM from JEOL or the same from Fuji) and CCD cameras (for example from Gatan or another companies) in 1,000 kV TEM?
Thank you in advance, Victor Sidorenko, ANTRON Co., Ltd., scientific service, Moscow, Russia.
Do any of the more geologically-inclined persons have an e-address for the company that makes the Franz magnetic separator?
thanks
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear Chris, If you check our new website, em-preparation.com you can download our manual, 'AFS Recipe book'. Here you will find a chapter on the freeze substitution of plant tissue including leaves. I hope you find it useful. Regards,
Ian Lamswood Marketing Manager, EM Products Leica, Vienna
Having run a listserv I can appreciate the stable service that Nestor provides and appreciate the quality assistance the list provides. I have seen it found many things that I though were made of pure unobtainium with ease and help the list provides people world wide.
This is the best all around list I follow and I follow a lot. I don't have a lot to do being retired and not to mobile. I have a broad range of interests in science and agriculture and there is no question that Nestor and you members provide the best product out there. Be it a Nobel laureate or a 6th grader you treat their question with consideration and usualy present several approaches.
You should be presented a pin at the next meeting and clusters with each additional billion messages or a pitcher of beer your choice.
} From all of us the appreciate your work
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
} From: {zaluzec-at-sparc5.microscopy.com} Sent: Sunday, November 04, 2001 12:32 PM
Hi,
CMP Cientifica today published the first Nanotechnology Overview white paper, attempting to give a global overview of nanotechnology. Although written with a business audience in mind, there is probably plenty in there to interest anyone working at the nanoscale.
The white paper (1.5 Mb) is available for free download at www.cmp-cientifica.com & other locations.
Any feedback from the microscopy community would be much appreciated, feel free to contact me directly.
Regards
Tim
Disclaimer: The full version of the Report is a commercial product, but the White Paper is freely available at no charge.
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
I'd like to add a suggestion to the one's by Matt listed below:
} } I'm usually wary when someone is fixated on a particular } "magic" number, 30 or 300 or whatever. Often you are } dealing with a person who doesn't understand statistics } nearly as well as they think they do. Since you haven't } been able to convince her of your position, maybe you can } ask her to justify hers? Make her really explain (and } hopefully understand) where that 30-40 target comes from. } I don't necessarily *recommend* this approach, since it } may lead to more conflict than collaboration!... } } Matt
} } } } My addition; If this person is so hung up on doing 30-40 } } } } samples, peerhaps you can give him'her instructions n sample prep } } } } and then tell him'her that s/he should feel free to run up all } } } } the samples on his/her time and that you would then gladly 'scope } } } } them. I have found that when people learn by personal } } } } experience how long it takes to do even a "standard' TEM } } } } embedding they suddenly readjust their "n" requirement to a more } } } } realistic goal.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
A customer of mine has a Kevex Delta II analyzer (circa 1989) equipped with the 8 inch/10 Mb Bernoulli disk drives. He would like to continue using this media for the time being but is unable to locate any of these disks for data storage. Thermo Noran is apparently unable to upgrade his drives to something more current.
If anyone has these disks laying around, and would like to sell them, please contact me off-line. Thank you.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 480.967.3946 "Home of the 2001 World Series Champion Arizona Diamondbacks"
Don't see what you need? Don't worry! We'll make it!
GaAs Stock (More Available, including 4") 1103SACON 76.2 mm x 635 um (100)SI,undp R} 3x107 EPD {7 x 104 2Pol Epi=09 1104SACON 76.2 mm x 635 um (100)SI,undp R} 2x107 EPD {7x104 2Pol Epi E-J Fl= ts=09 A0244 20x25mmx381um Zn HB cc=3D1018, (111) 1pol/1etch, EPI, EPD {104=09 AR250E 76.2 mm dia. (100) Semi Insul, 2 x 108 ohm-cm, As-Cut=09 A0432 2=94dia x 450 um (100) N/Te, cc=3D1-5 x 1017, 2pol, EPD {8x104=09 A0437 2=94dia x 450 um (100) P/Zn, cc=3D1-5 x 1017, 2pol, EPD {8x104=09 A0860 76.2 mm x 600 um (100) N/Te cc=3D3 x 1018, 1 Pol, EPI=09 A1291 50.8 mm x 450 um (111) P/Zn, cc=3D1-2 x 1017 EPD:3-5x104 , 2 pol Ep= i=09 A1397 76.2 mm x 600 um (100) SI, Undp R: 1-2 x 108 EPD {8x104, 2 pol, Epi=09 A1583 76.2 mm x 600 um (111) N/Te, cc} 2 x 1018, EPD {8x104, 1 pol, Epi=09 A1595 76.2 mm x 600 um (111) N/Te, cc} 1 x 1018, EPD {8x104, 1 pol, Epi=09 A1784 50.8 mm x 375 um (111) N/Si, cc} 1 x 1017, EPD {5x104, 1 Pol, Epi=09 L6049 76.2 mm x 600 um (100) P/Zn, cc=3D1-5 x 1017, EPD { 8 x 104, 2 pol,= Epi=09 L6097 50.8 mm x 400 um (110) SI, Undp R} 1x108, EPD {8x104, 2pol, Epi=09 L6133 50.8 mm x 400 um (111)Ga, SI, Undp R} 1x107, EPD {8x104, 1pol, Epi=09 L6165 50.8 mm x 400 um (100), SI, Undp R} 1x107, EPD {8x104, 2 pol, Epi=09 L6223 50.8 mm x 450 um (100) P/Zn, cc=3D2-4 x 1018, EPD { 8 x 104, 1 pol,= Epi=09 ---------
GaP L6114 50.8 mm x 300 um (111) N/S, cc: 1 - 5 x 1017 EPD { 2 x 105 2 pol EP= I=09 L6208 50.8 mm x 300 um (100),N/Undpd,cc { 1 x 1016,EPD { 2 x 105,2Pol EPI= =09
------------
GaSb 11108SA 2=94 2 Polished=09 1.75=94 (100), P-Type =09 2=94 x 4 mm (100), N/Te, Blanks, R { 1017, EPD { 104, Unpol=09 L5467 50.8 mm x 500 um (100), N/Te, cc =3D 6-8x1017, EPD { 1x103, 1 pol=09 L6116 50.8 mm x 500 um (100) P/Undpd, cc:1-2 x 1016,EPD {5 x 103, 1 Pol Ep= i=09
------=20 InAs A0254 50.8 mm x 450 um Zn/P-Type (100) 4o to(111), cc:2x1018, 1 pol/1 etc= h=09 A0300 50.8 mm x 450 um S/N-Type (100) 4o to(111)A, cc: 1018, 1 pol/1 etch= =09 A0352 50.8 mm x 450 um Zn/P-Type (100)2o to(110), cc:5-7x1018, 1pol, Epi=09 A0358 50.8 mm x 450 um S/N-Type (100) 4o to(111)A, cc: 3-5x1018, 1 pol=09 A0359 50.8 mm x 450 um S/N-Type (100) 4o to(111)A, cc:3x1018, 1 pol/1 etc= h=09
------- =09 InSb L5728 50.8 mm x 450 um (111)A, N/Undpd cc: 4-7x1014, EPD {200, 2 pol, Epi= =09 L5885 50.8 mm x 450 um (100), N/Te, cc: 3-5x1017, EPD {200, 1 pol, Epi=09 L6207 50.8 mm x 450 um (100), N/Undpd, cc: 4-7x1014, EPD {200, 2 pol, Epi= =09
-------- =09 InP 50.0 mm x 500 um Prime, SI/Fe,(100),R} 1x107,EPD {5x104,E-J,1pol,Epi=09 50.0 mm x 500 um Test, SI/Fe,(100), R} 1x107,EPD {5x104,E-J, 1pol,Epi=09
InP Indium Phosphide, LEC (100) +/- 0.2 degrees Orientation Semi Insulating, Iron doped Resistivity } 1 x 10^7 Ohm cm EPD { 5 x 10^4 cm^-2 Euro-Japanese Flats 1 Side Polished, Back Lapped and Etched 50.0 mm diameter x 500 microns thick
-----------------
InGaAs Wafer 2"=D8 with InGaAs EPI on SI InP {100} , by MOCVD deposition=20 Substrate: Semi-Insulating InP (eg. InP:Fe), Resistivity: } 1 x 10E7 Ohm = cm, EDP { 1 x 10E4/cm2)EPI: L attice matched In/Ga alloy layer of n-type InGaAs Nc } 2 x 10E18/cc (usin= g Si as dopant) Thickness: 0.5 um (+20%)
Ultra-Thin wafers now in-stock as is Float Zone and Ultra-flat wafers.
We also have fresh stocks of 2"-6" GaAs, GaSb, GaP Germanium, InP, InAs, = InSb, Al203, ZnO, ZnSe and more! They're ready to ship!
I'll forward this to the Classic Computer Collector's mailing list. Someone there certainly will have a lead on some of these.
- John
At 10:18 AM 11/5/01 -0700, Bob Roberts wrote: } A customer of mine has a Kevex Delta II analyzer (circa 1989) equipped with the 8 inch/10 Mb Bernoulli disk drives. He would like to continue using this media for the time being but is unable to locate any of these disks for data storage. Thermo Noran is apparently unable to upgrade his drives to something more current. } } If anyone has these disks laying around, and would like to sell them, please contact me off-line. Thank you. } } Bob Roberts } EM Lab Services, Inc. } 2409 S. Rural Rd Suite C } Tempe, Arizona 85282 } 480.967.3946 } "Home of the 2001 World Series Champion Arizona Diamondbacks"
----- Original Message ----- } From: "Leona Cohen-Gould" {lcgould-at-med.cornell.edu} To: {mlynn-at-miami.edu} ; {microscopy-at-sparc5.microscopy.com} } } } } Hi Paula, } } } I'd like to add a suggestion to the one's by Matt listed below: } } } } } I'm usually wary when someone is fixated on a particular } } "magic" number, 30 or 300 or whatever. Often you are } } dealing with a person who doesn't understand statistics } } nearly as well as they think they do. Since you haven't } } been able to convince her of your position, maybe you can } } ask her to justify hers? Make her really explain (and } } hopefully understand) where that 30-40 target comes from. } } I don't necessarily *recommend* this approach, since it } } may lead to more conflict than collaboration!... } } } } Matt } } Have someone that really understands statistics look at the paper. There is no magic number it all depends on the varibility of the data and what you are saying about the data.
I doesn't sound like either of you can make a strong case for your position. At least not one that convinces the other.
Good luck Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
----- Original Message ----- } From: "John Foust" {jfoust-at-threedee.com} To: "Bob Roberts" {bobrobs-at-earthlink.net} ; {Microscopy-at-sparc5.microscopy.com} Cc: {classiccmp-at-classiccmp.org} Sent: Monday, November 05, 2001 7:49 PM
I might be willing to share some of my 10 MB Bernoulli cartridges. Since I'll be holding onto some of the drives, I expect I'll not let them all go.
I do have 20 MB cartridges and drives as well, and those drives read the 10 MB media, yet write only the 20's. At one time it was convenient to put a bernoulli box at each of several clients' locations in order simply to move the cartridges along with me, yet be able to accomplish equivalent useful work at home, which has always been my primary work location.
regards,
Dick
----- Original Message ----- } From: "John Foust" {jfoust-at-threedee.com} To: "Bob Roberts" {bobrobs-at-earthlink.net} ; {Microscopy-at-sparc5.microscopy.com} Cc: {classiccmp-at-classiccmp.org} Sent: Monday, November 05, 2001 7:49 PM
Maybe..
} } } Arthur Day {ard-at-ansto.gov.au} 11/06/01 02:50PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It would be easier to send Nestor to where the beer is. Less postage that way...
} } I say we all mail Nester one beer, something regional or unique. Just } picture the scene :-) } } } -Ed
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
Tell you what - If all the List subscribers come here I can promise the Hawaii Visitors Bureau will buy you ALL a beer! And 2,562 for Nestor ...
Mahalo nui loa, Nestor!
Tina
http://www.pbrc.hawaii.edu/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Ok Sally I admit that last message was a bit ambiguous. I'll blame the Melbourne Cup BBQ for that (beer). So how do I weasel out of it?
Nestor wrote: "Now if I only had a beer for each of those emails, I'd be set for life."
If nature abhors a vacuum/empty stomach, and if there is no such thing as one beer, then each of us may have to contribute up to 100,000,000 beers each -- one per message. That's up to 100,000,000 times 2562. So, therefore, it would be easier to send Nestor to each exotic location to sample the beer in person?
Bit of an extrapolation I suppose.
If anybody needs some tips for their next grant proposal.... ;-(
} Maybe.. } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I saw a hand full of this media that VCF East...Did they get sold? I'm not sure who the seller was...maybe Tom Owad? -Chandra
-----Original Message----- } From: owner-classiccmp-at-classiccmp.org [mailto:owner-classiccmp-at-classiccmp.org] On Behalf Of John Foust Sent: Monday, November 05, 2001 9:50 PM To: Bob Roberts; Microscopy-at-sparc5.microscopy.com Cc: classiccmp-at-classiccmp.org
Having mis-spent my youth at Lehigh University, I suggest we all HAVE a beer FOR Nestor!
Fred Monson
} ---------- } From: "Edward_Principe-at-amat.com"-at-sparc5.microscopy.com } Sent: Monday, November 5, 2001 7:59 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Beer fund } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I say we all mail Nester one beer, something regional or unique. Just } picture the scene :-) } } } -Ed } } }
The web page for Franz is http://www.sgfrantz.com/
Sincerely, James Talbot
K/T GeoServices, Inc. Bulk and Clay Mineralogy by X-ray Diffraction (214) 403-6342 web site: http://www.ktgeo.com/
Ritchie Sims wrote:
} } Hello world } } Do any of the more geologically-inclined persons have an e-address } for the company that makes the Franz magnetic separator? } } thanks } } rtch } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
} Kristen Lennon wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Semantics because this mistake has been making me crazy - ANTHRAX IS A } } BACTERIUM! This is a particularly important point because the word "virus" } } scares people silly. When I think of virus, I think of AIDS and Ebolla } } (sorry it that's misspelled). When I think of bacterium, I think antibiotic. } } No offense intended, } } Kristen } } } } Kristen A. Lennon, Ph.D. } } Department of Plant Pathology } } 351 Bessey Hall } } Iowa State University } } Ames, IA 50011 } } } } 515-294-8854 } } kalen-at-iastate.edu } } www.baumlab.org } } -- } _____________________________________________________ } Tyrone L. Daulton } Director: Marine Geosciences - Electron Microscopy Center } Physicist and Senior Electron Microscopist } } Naval Research Laboratory } Marine Geosciences Division (Code 7400) } Stennis Space Center, MS 39529-5004 } } Voice (228)-688-4877 } Fax (228)-688-4093 } email tdaulton-at-nrlssc.navy.mil } tld-at-howdy.wustl.edu }
Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was? I've come across a reference to it in a 1982 paper by Eddy and Koehler and need to describe the SEM imaging technology for something I'm writing.
Thanks in advance for any help.
Best,
Angela
Angela V. Klaus
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
I think someone had once posted a URL to a website with an HTML presentation that moved the observer from the sub-atomic viewing level to one outside of our solar system using metric increments. Could someone please send me that URL again?
I still say it would be easier to have a sampling of beer, and, Oh, alright, tell Nestor how good it (OR THEY!!!!) WAS!!!!
Fred Monson I'm heading out now to start!!! See all of you who are down-time from me, later!!! } ---------- } From: Arthur Day } Sent: Tuesday, November 6, 2001 2:43 AM } To: Sally Stowe } Cc: microscopy-at-sparc5.microscopy.com; Edward_Principe-at-amat.com } Subject: Re: Beer fund } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ok Sally I admit that last message was a bit ambiguous. I'll blame } the Melbourne Cup BBQ for that (beer). So how do I weasel out of it? } } Nestor wrote: "Now if I only had a beer for each of those emails, } I'd be set for life." } } If nature abhors a vacuum/empty stomach, and if there is no such } thing as one beer, then each of us may have to contribute up to } 100,000,000 beers each -- one per message. That's up to 100,000,000 } times 2562. So, therefore, it would be easier to send Nestor to each } exotic location to sample the beer in person? } } Bit of an extrapolation I suppose. } } If anybody needs some tips for their next grant proposal.... ;-( } } } } Maybe.. } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It would be easier to send Nestor to where the beer is. Less postage } } that way... } } } } } } } } I say we all mail Nester one beer, something regional or unique. Just } } } picture the scene :-) } } } } } } } } } -Ed } } }
Comments to comments: } } I'm usually wary when someone is fixated on a particular } "magic" number, 30 or 300 or whatever. Often you are } dealing with a person who doesn't understand statistics } nearly as well as they think they do. Since you haven't } been able to convince her of your position, maybe you can } ask her to justify hers? Make her really explain (and } hopefully understand) where that 30-40 target comes from. } I don't necessarily *recommend* this approach, since it } may lead to more conflict than collaboration!... } } Matt
} } } } My addition; If this person is so hung up on doing 30-40 } } } } samples, peerhaps you can give him'her instructions n sample prep } } } } and then tell him'her that s/he should feel free to run up all } } } } the samples on his/her time and that you would then gladly 'scope } } } } them. I have found that when people learn by personal } } } } experience how long it takes to do even a "standard' TEM } } } } embedding they suddenly readjust their "n" requirement to a more } } } } realistic goal. Lee
Reply: Unfortunately (or fortunately), the actual sample number is not arrived at in an arbitrary manner, it is dictated by the nature of the specimen under study.
Nobody makes up these numbers, but if you look at the article by John Lucocq, Trends in Cell Biology 1993 Unbiased 3-D quantitation of ultrastructure in cell biology 3:354-358, he suggests using 3-5 experiments/animals/cultures, only 1 or more blocks/dishes, one or more sections but 10-40 micrographs and between 100-200 probe counts. I am pretty sure this is a common sampling scheme for stereologists.
Paraphrasing slightly form this manuscript, Sampling is "..an inherent part of any quantitative electron micrscopy study since only a portion of all the possible blocks, sections and micrographs are analyzed. It is essential therefore that at each level in a sampling scheme items are selected in a way that gives all parts of the specimen the same chance of being included."
The actual amount of sampling that is performed will be directly related to the variance occurring within the population. Samples with low variance will require a lower sample number to give you an answer with an error of 5% or less. High variance between individual samples will require more sampling to be performed.
These are all experimental design considerations that have to be applied much before all the clever statistical analysis is performed. Working out probabilities on poorly sampled data is not very useful.
It can be said that proper sampling techniques should also be applied to morphological data too. Where would that put all those text-book images?
AIDS is a name of the illness caused by VIRUS, I believe. Virus itself has a name: HIV, Human Immuno-deficit Virus. Sergey
At 10:36 AM 11/6/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Yes, Ebola is a virus, AIDS is caused by a virus and Anthrax is caused by a bacterium. How come I, as a lowly physicist, know this but others do not, although they take great delight in trumpeting their ignorance for all of us to see!
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } AIDS is a name of the illness caused by VIRUS, I believe. Virus } itself has a name: HIV, Human Immuno-deficit Virus. } Sergey } } At 10:36 AM 11/6/01, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Note new cell phone number!
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
I'm a bit of a novice with antibody staining, but so are my colleagues and so they've all got conflicting and inconsistent results from trying the Alkaline Phosphatase staining.
We're trying to stain sea urchin larvae whole-mounts with the 2 antibody-alkaline phosphatase system(larvae are formalin fixed and rehydrated from 70% EtOH cold storage). Our AkPh substrate is a few months old and has a small amount of blackish precipitate at the bottom--is this OK? My colleagues have had different results with using the substrate undiluted on the animals vs. diluted in some small amount of PBS. I notice that with the latter, there is a slight whitish precipitate that occurs--is this due to the pH differential between the AkPh substrate(ph 10 or more) and PBS(7.4)? What are the ideal staining conditions for this to work? Please note--I'm getting problems with the preliminary staining where I know I'm supposed to get endogenous gut AkPh staining with just substrate and nothing else, but I'm getting almost imperceptible staining.
All suggestions and admonishments will be taken into consideration. Thanks in advance.
Pauline Yu pcy-at-usc.edu Manahan Research Laboratory http://www.usc.edu/manahanlab University of Southern California
There's a high school teacher in Hull, Massachusetts with 20 or so microscopes that need cleaning and (hopefully minor) repair. She's willing to work on them herself, but doesn't know how to begin. Is there a lister who can visit and help her get started? If so please contact me.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
on 11/4/01 4:39 PM, Victor Sidorenko at antron-at-space.ru wrote:
} } Does anybody have the information about the using possibility of } image recording systems (like PIXsysTEM from JEOL or the } same from Fuji) and CCD cameras (for example from Gatan or } another companies) in 1,000 kV TEM? } Dear Victor, A few years ago (when the 1250 kV scope had been installed at Stutgart, but I don't remember the exact year) there was a poster at MSA which gave the quantum efficiency for image plates as a function of voltage up to 1250 kV. Since I was interested in quantitating ED data at 1200 kV, and since I had heard great things about IPs--single-electron sensitivity, linearity for a range which would include all the expected intensities in an ED pattern, etc.--I remember the graph well. The QE for somewhat more than 100 kV was 100% (giving the required sensitivity), but it dropped off at intermediate voltages--I don't remember whether it was still 100% at 200 kV, but definitely lower at 400 kV--and was only ~25% at 1000+ kV. I don't know what improvements may have been made; a thicker active layer will give more sensitivity, but lower spatial resolution (there is no free lunch). The manufacturers of the plates would be able to tell you whether they can sell you what you would need. Good luck. Yours, Bill Tivol
ACEM 17 The 17th Australian Conference on Electron Microscopy Adelaide Convention Centre, Adelaide, South Australia February 4 - 8, 2002 Hosted by Australian Society for Electron Microscopy (Inc.)
This meeting will provide an opportunity to explore the many applications of Electron, Scanned Probe and Confocal Microscopies, to keep abreast of the latest developments and to hear and interact with many of the leaders in the field. Sessions on Cryotechniques, Virology/Microorganisms, Cell Tomography, Nanotechnology, Emerging EM Technologies, Materials and Microstructure, Superresolution and an Industry Forum are being planned, among many others.
Invited and Keynote Speakers (to date) Martin Bootman; Confocal (biological), David Cockayne; Nanostructure, Patrick Echlin; Analytical Microscopy of Biomaterials, Mark Fricker; Confocal (biological), David Jefferson; Emerging Techniques, Jan Leunissen; Immunolabelling, Ohad Medalia; Cellular Tomography, Greg Meeker; Microbeam Analysis, Sara Miller; Virology, Daniel Mueller; SPM - (biological), David Williams; Microanalysis, Nestor Zaluzec; Analytical Microscopy/Telepresence Jamie Chapman; Reproductive Biology, Jacques Dubochet; Cryotechniques, Alex Hyatt; Virology/Cryotechniques, Sergei Magonov; SPM, Mary, Mah-Lee Ng; Virology/Confocal
A series of pre-conference workshops will be held to allow participants to gain hands-on experience in the latest techniques and applications. Many of the invited and keynote speakers will participate as presenters, providing a wonderful opportunity for interaction with these noted scientists. Be sure to book soon, as places in some of the workshops are limited.
Workshops Cryo Scanning Electron Microscopy Environmental Scanning Electron Microscopy, Techniques and Applications Image Analysis Leica-Aurion Workshop on Cryotechniques and Immunolabelling Light Element Analysis: Problems and Solutions Quantitative Confocal Microscopy Scanning Probe Microscopy Trace and Major Quantitative X-ray Mapping (a Hands on Approach) Virology Using Electron & Light Microscopy
During the week, delegates will have the chance to mingle and relax after hours in several of Adelaide's noted heritage locations. A range of accommodation choices, from five-star to budget, ensures choices suitable for all.
The Trade Exhibition will be a comprehensive and exciting display of the latest equipment and accessories for microscopy, imaging and analysis. We are most grateful to our sponsors and exhibitors for their very generous support of ACEM17.
As is traditional at all ACEMs, Poster and Micrograph Competitions will be run. Categories for the Poster Competition will be Best Biological Poster, Best Physical Poster and Best Student Poster. Categories for the Micrograph Competition are SEM, TEM, SPM and Confocal. Entries will be judged on aesthetic, photographic and scientific merit. Entry details are available on the website.
With the deadline for abstract submission fast approaching, (November 30), make plans now to join us in February! (Early bird registration deadline - November 16). Abstracts should be submitted electronically, and the registration form can be downloaded from the website - visit often for regular updates.
We look forward to seeing you at ACEM17.
Full details can be obtained at http://www.adelaide.edu.au/CEMMSA/acem17
The JEOL 100CX TEMSCAN is a 100kV transmission electron microscope (TEM) with the attachment for scanning imaging device (ASID) that provides scanning electron microscope (SEM) facilities (the SCAN part of TEMSCAN). It could image with secondary electrons (SE) or backscattered electrons (BSE) in a standard SEM mode or use brightfield or darkfield detectors for scanning transmitted electron microscopy (STEM). With an energy dispersive x-ray (EDX) specrometer fitted it could also provide chemical analysis, x-ray maps and line scans.
Undoubtably there are still some out there being used and their users could give you more information but your local JEOL office could also give you any information you want (ask for an engineer over 40). If you don't know the local JEOL office look on the web, although they maybe rushing out to you now in case you want to buy something!!!
Regards, Ron
On Tue, 6 Nov 2001 14:00:30 -0500 (EST) Angela Klaus {avklaus-at-amnh.org} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all... } } Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was? } I've come across a reference to it in a 1982 paper by Eddy and Koehler and } need to describe the SEM imaging technology for something I'm writing. } } Thanks in advance for any help. } } Best, } } Angela } } Angela V. Klaus } } Director, Core Imaging Facility } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024 USA } } Email: avklaus-at-amnh.org } Tel: 212-769-5977 } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
The 100CX TEMSCAN is a JEOL 100kV TEM fitted with a scanning attachment which allows imaging by way of "in lens" surface studies (SEM) in addition to transmitted electron scanning (STEM). The microscope was a second generation 100C hence the X.
The instrument was designed by Yanaka and Shirota the scanning system interface was made by Koike (the man who invented the in lens SEM system). Koike went on to design the ISI DS130 SEM and Yanaka and Shirota the ISI 002B TEM.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
The 100CX TEMSCAN is a JEOL 100kV TEM fitted with a scanning attachment which allows imaging by way of "in lens" surface studies (SEM) in addition to transmitted electron scanning (STEM). The microscope was a second generation 100C hence the X.
The instrument was designed by Yanaka and Shirota the scanning system interface was made by Koike (the man who invented the in lens SEM system). Koike went on to design the ISI DS130 SEM and Yanaka and Shirota the ISI 002B TEM.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Angela Klaus wrote: Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was?
If we're talking about the same instrument, the JEOL 100 CX that I spent years on was a TEM (100 kV with a booster switch to 120 kV accelerating voltage). Mine had the SAED (selected area electron diffraction) accessory and Scanning EM functions.
Coincidentally, I know where just such an instrument is available for the cost of tear down and shipping to any federal institution OR an institution with an active NSF grant using electron microscopy. It includes a full-sized, pneumatic anti-vibration table, chiller, compressor, film carriers, etc.
Contact me off-line if interested and I'll see if it's still available.
S. Frank Platek Forensic Chemistry Center U.S. Food and Drug Administration 6751 Steger Drive Cincinnati, OH 45237-3097 (513) 679-2700 X254 (513) 679-2761 FAX fplatek-at-ora.fda.gov
We are interested in doing EELS work with a LaB6 filament TEM. Can anyone tell us how best is the energy resolution with this type of filament?
Thanks
Best Regards Yan Xin ======================================= Yan Xin (Ph.D) Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
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That would be "Powers of 10", by Eames. I know it as a video, book, and flip book (see the MICRO bibliography, URL below), but not as a website. Will you please post the URL on the list when you get it?
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Absolutely correct John! And, since you know, I'll ask.
Which physicist is most responsible for "nucular" physics [did I spell it right???], and which Computer Scientist for "data is"?
I know the biologists are responsible for "dIsection"!
Blessings upon the lexicographers who say, "Cut it any way you want! DIsect it with a cleaver or DISsect it with fine forceps. Your choice, whether you know the difference or not!"
As Grandfather used to say when Grandmother was near, "Oh! For the love of words!!!"
Fred Monson, who hath some affection for the silly!
P.S. By the way, Nestor, I stopped on the way home last evening, tried a few different brands to be found in Eastern Pennsylvania, just Northwest of Philadelphia, and most were worth the try. Every one went down as a toast to your good works, and that was good too! So, you are taken care of here, and I hope you appreciate the effort on your behalf. FCM
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: John F. Mansfield } Sent: Tuesday, November 6, 2001 9:12 PM } To: microscopy-at-microscopy.com } Subject: Virus and Bacterium } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Yes, Ebola is a virus, AIDS is caused by a virus and Anthrax is } caused by a bacterium. How come I, as a lowly physicist, know this } but others do not, although they take great delight in trumpeting } their ignorance for all of us to see! } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } AIDS is a name of the illness caused by VIRUS, I believe. Virus } } itself has a name: HIV, Human Immuno-deficit Virus. } } Sergey } } } } At 10:36 AM 11/6/01, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } AIDS isn't a virus either. More semantics. } } } } } } } } } } Kristen Lennon wrote: } } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } Semantics because this mistake has been making me crazy - ANTHRAX IS } A } } } } } BACTERIUM! This is a particularly important point because the word } "virus" } } } } } scares people silly. When I think of virus, I think of AIDS and } Ebolla } } } } } (sorry it that's misspelled). When I think of bacterium, I think } } } antibiotic. } } } } } No offense intended, } } } } } Kristen } } } } } } } } } } Kristen A. Lennon, Ph.D. } } } } } Department of Plant Pathology } } } } } 351 Bessey Hall } } } } } Iowa State University } } } } } Ames, IA 50011 } } } } } } } } } } 515-294-8854 } } } } } kalen-at-iastate.edu } } } } } www.baumlab.org } } } } } } } } -- } } } } _____________________________________________________ } } } } Tyrone L. Daulton } } } } Director: Marine Geosciences - Electron Microscopy Center } } } } Physicist and Senior Electron Microscopist } } } } } } } } Naval Research Laboratory } } } } Marine Geosciences Division (Code 7400) } } } } Stennis Space Center, MS 39529-5004 } } } } } } } } Voice (228)-688-4877 } } } } Fax (228)-688-4093 } } } } email tdaulton-at-nrlssc.navy.mil } } } } tld-at-howdy.wustl.edu } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } -- } Note new cell phone number! } } Dr. John Mansfield CPhys MInstP } North Campus Electron Microbeam Analysis Laboratory } 417 SRB, University of Michigan } 2455 Hayward, Ann Arbor MI 48109-2143 } Phone: (734) 936-3352 FAX (734) 763-2282 } Cellular Phone: (734) 834-3913 } (Leaving a phone message at 936-3352 is preferable to 834-3913) } Email: jfmjfm-at-engin.umich.edu } URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html } Location: Lat. 42° 16' 48" Long. 83° 43' 48" } }
Does anyone know a recipe for chemically etching GaN and/or InN? The material is already thinned and ready for TEM, we just want a final surface etching to enhance dislocation imaging.
Thanks,
Mick
------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Angela Klaus } Sent: Tuesday, November 6, 2001 2:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Info needed: JEOL 100 CX TEMSCAN } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all... } } Does anyone know what type of microscope the JEOL 100 CX TEMSCAN is/was? } I've come across a reference to it in a 1982 paper by Eddy and Koehler and } need to describe the SEM imaging technology for something I'm writing. } } Thanks in advance for any help. } } Best, } } Angela } } Angela V. Klaus } } Director, Core Imaging Facility } American Museum of Natural History } Central Park West at 79th Street } New York, NY 10024 USA } } Email: avklaus-at-amnh.org } Tel: 212-769-5977 } } }
} -----Original Message----- } From: Sy Griffey } Sent: Wednesday, November 07, 2001 11:48 AM } To: Jacqueline D. Garfield } Subject: Processing of tendon samples } } } } If anyone has some experience processing human tendon samples for EM } and/or histology we would appreciate your help. } } Problem: We have begun processing human tendon samples for both EM and } standard histology but are having trouble getting good sections and tissue } quality. The samples for histology can be as thick as 5-8 mm. } } For the EM samples we have only done a freeze-substitution protocol } to date but the tissue was extremely hard before embedding and even thick } sectioning was a challenge. We anticipate similar problems when doing } standard EM processing based on experience with histology processing thus } far........The tissue in the histology sections appears cracked, } exhibiting separation within the fibrous bundles of the tendon when viewed } in X-sxn. The samples were actually somewhat flexible just prior to } embedding. All processing steps (graded alcohol, clearing solution and } paraffin) have been done at ambient temperature and pressure. The } incubation times, in general, have been in the 45-60 min. range. We are } thinking about increasing the incubation times and perhaps doing some } vacuum infiltration but would welcome any suggestions. } } Thank you! } } Jackie Garfield and Sy Griffey } } Lifecell Corporation } One Millennium Way } Branchburg, NJ 08876 USA } (908) 947-1182 (Jackie) } (908) 947-1085 (Fax) } jgarfield-at-lifecell.com
Thank you for the many replies regarding the problem of firmly-attached carbon.
It appears that in removing the metal coating left on the glass surface in the bell jar's previous life in a physics lab, the aqua regia that I ended up using activated the glass surface, perhaps by producing a micro-roughness. So the first few uses produced a firmly-attached carbon coating.
I was able to remove the carbon with a mildly-abrasive ("without harsh scratching") household cleaner, plus lots of rubbing.
I then wiped the surface with a rag wetted with kerosine, then with a clean dry rag. This left sufficient kerosine to prevent a good vacuum being obtained, so I sprayed it with a household general-purpose multi-solvent cleaner ("Ajax Spray 'n' Wipe), and wiped that off with a clean dry rag.
This has left the surface sufficiently deactivated so that I can remove new carbon simply by spraying with Spray 'n' Wipe and wiping.
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
A graduate student in microbiology has just finished her first course in electron microscopy, and she informs her advisor that she wants to study mitochondrial structure and function in several related strains of yeast, for one of which she fortuitously has electron micrographs of mitochondrial profiles that are, she has been told, quite publishable.
Her advisor suggests that she start by comparing 3-4 strains with respect to number of mitochondria per cell and total mitochondrial cristal area under identical conditions of culture.
The student happily responds that she already has micrographs for regularly sampled sections from serial sections she learned to make of the strain she used in her EM course. She returns the next week with estimates of cristal area and numbers of mitochondria per cell based on morphometric analysis of those regularly sampled sections.
If the cell is 12-25um in length (haploid and diploid) and always between 3-5um in diameter, and if the mitochondrial profiles are in the range of 1-2um (circular and ovoid, and sometimes or often branched), what should the sampling criteria be for cells that are serially sectioned with sections approximately 90nm in thickness?
How could one derive the same, but apparently much more statistically satisfactory, data from the 40+ cell profiles per section of cells randomly oriented and not necessarily (or importantly) sectioned or analyzed completely.
All who take on this task should know there is an extremely important, and possibly significant, hint that can be observed at: http://www.msg.ucsf.edu/sedat/marsh/mito.html. I sometimes think it is well to know of such hints (or at least to be aware of their possibility) before attempting a task. Assumptions are almost always incorrect unless rigorously defined and applied, and very often difficult to prove as such or to isolate in generating an experimental design without flaws.
Knowing that reality often gets in the way of statistics, and vice versa, I am
Yours most respectfully, because I wrestle with this problem every once in a while and I haven't yet determined a satisfactory answer.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I just looked in Yahoo and found the website. It's www.powersof10.com.
Edward Haller University of South Florida Pathology Department Tampa, Florida
Caroline Schooley wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear All, } } } } I think someone had once posted a URL to a website with an HTML } } presentation that moved the observer from the sub-atomic viewing level to } } one outside of our solar system using metric increments. Could someone } } please send me that URL again? } } } } Thanks, } } } } Louis - } } That would be "Powers of 10", by Eames. I know it as a video, book, and } flip book (see the MICRO bibliography, URL below), but not as a website. } Will you please post the URL on the list when you get it? } } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Can anyone in the DFW area perform these types of analysis? Respond to me with details and contact information.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
-----Original Message----- } From: Rajan Vempati [mailto:rvempati-at-mail.smu.edu] Sent: Wednesday, November 07, 2001 10:16 PM To: rbeavers-at-mail.smu.edu
I am looking for a simple, reliable method for obtaining thin sections for EM from thick (1-2 microns) epoxy resin sections that have been cut, mounted on slides, and stained for preliminary examination with the LM .
Hi All, After Caroline's message I did a web search for "powers of 10" and found this web site which seems to fit what was requested. http://www.wordwizz.com Steve
Steve Buckingham Dir. of Process Development Excellatron Solid State LLC 1640 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
} -----Original Message----- } From: Boche, Jo Ellen } Sent: Wednesday, November 07, 2001 4:33 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: TEM Request for method for cutting thin sections from } thick. } } } } I am looking for a simple, reliable method for obtaining thin sections } for EM from thick (1-2 microns) epoxy resin sections that have been } cut, mounted on slides, and stained for preliminary examination with } the LM. } } bochej-at-boystown.org } } Jo Ellen Boche } Neuroanatomy Laboratory } Boys Town National Research Hospital } Omaha, NE 68131 } (402) 498-6531 } } .
You can "just" re-embed the sections directly on the slide (invert the slide-bound sections on BEEM capsules filled with resin or put the capsule on the slide), repolymerize, and cut. I always manage to make a serious mess with the resin, but it does work.
You may also be able to glue the thick section to a polymerized blank block and then pop it off the slide with a heat/cold regimen - I haven't tried this but it is supposed to work. I think it may be more useful/less disastrous with a REALLY thick section than a normal thick section.
Good luck!
Tamara
On Wed, 7 Nov 2001, Boche, Jo Ellen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } } -----Original Message----- } } From: Boche, Jo Ellen } } Sent: Wednesday, November 07, 2001 4:33 PM } } To: 'Microscopy-at-MSA.Microscopy.Com' } } Subject: TEM Request for method for cutting thin sections from } } thick. } } } } } } } } I am looking for a simple, reliable method for obtaining thin sections } } for EM from thick (1-2 microns) epoxy resin sections that have been } } cut, mounted on slides, and stained for preliminary examination with } } the LM. } } } } bochej-at-boystown.org } } } } Jo Ellen Boche } } Neuroanatomy Laboratory } } Boys Town National Research Hospital } } Omaha, NE 68131 } } (402) 498-6531 } } } } . } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
For this reason it's more correctly to call it: "mitochondrion" (M), instead mitochondria. In my point of view, on EM level only 3D reconstruction from the serial sections may give you truthful info about this structure.
By the way, mitochondria/mitochondrion is known as an extremely sensitive organelle: to oxidative stress etc&etc. Its structure may be changed by very slight variation of condition/procedure. Simple example: you are looking how some particular mutation affected M structure. It's very possible, that your mutation will affect the membrane permeability nor M at all. On EM level this may cause the different conditions during sample fixation/processing. Finally you will detect M changes, because the processing conditions were different for control and mutant. Another example: mutants are growing slowly at many instances, it's very difficult to have the culture's "age" similar. People usually grow cells to the some particular optical density (at 450 nm?). So the control will grow up to 1 OD, let say 5 h, but the mutant - 18+ h. Are the "age" of the cells the same? I guess, not. The structure of the M in the cells of different "age" may (and could) be different. So, we have to be careful with M as, from another hand, all EM stuff we have deal with. Sergey
At 12:53 PM 11/7/01, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (george_greg-at-msn.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, November 7, 2001 at 19:50:43 ---------------------------------------------------------------------------
Email: george_greg-at-msn.com Name: Greg George
Education: 9-12th Grade High School
Location: Pearl, Mississippi USA
Question: Dear Sir, I recently received, as a gift, a used lab grade microscope without a user manual. Three of the four (oil immersion) objectives are spring retractable. Does this feature mean that I can touch the slide surface without fear of damaging the lens? Thank you for your kind reply, Greg George
Powers of 10 is also available as a Java applet at http://micro.magnet.fsu.edu/primer/java/scienceopticsu/powersof10/ At this moment, the website is inaccessible to me so I cannot verify for sure. Best Wishes.
-----Original Message----- } From: Caroline Schooley [SMTP:schooley-at-mcn.org] Sent: Thursday, November 08, 2001 9:25 AM To: Louis Somlai Cc: Microscopy-at-sparc5.microscopy.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That would be "Powers of 10", by Eames. I know it as a video, book, and flip book (see the MICRO bibliography, URL below), but not as a website. Will you please post the URL on the list when you get it?
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
The first site mentions Eames and would be the wanted site. The second site too is very good and worth a long look. Its part of the molecular expressions site at Florida State Uni. The third site gives a power of ten perspective on computer data sizes. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Thursday, November 08, 2001 1:46 AM, Caroline Schooley [SMTP:schooley-at-mcn.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear All, } } } } I think someone had once posted a URL to a website with an HTML } } presentation that moved the observer from the sub-atomic viewing level to } } one outside of our solar system using metric increments. Could someone } } please send me that URL again? } } } } Thanks, } } } } Louis - } } That would be "Powers of 10", by Eames. I know it as a video, book, and } flip book (see the MICRO bibliography, URL below), but not as a website. } Will you please post the URL on the list when you get it? } } } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html }
on 11/7/01 10:46 AM, Yan Xin at xin-at-magnet.fsu.edu wrote: } We are interested in doing EELS work with a LaB6 filament TEM. Can anyone } tell us how best is the energy resolution with this type of filament? } Dear Yan Xin, As I remember it, the energy spread in a LaB6 filament is about 1-2 eV. The energy resolution obtainable in EELS is affected by factors other than the filament, however, and I do not know the best that can be obtained with your instrument. Yours, Bill Tivol
We are trying to section a tampon in to about five micrometers but any normal sample prep involves EtOH or some other fluid that will be absorbed. i was wondering if anyone knew a method to keep it in at its normal size but in a form that can be cut very thin thanks john
} ===== Original Message From "Monson, Frederick C." {fmonson-at-wcupa.edu} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
With effect from 1st November 2001 the Polaron Range of EM preparation instruments will change ownership from Thermo VG Scientific to Quorum Technologies Ltd.
Quorum Technologies Ltd. is the company formed by Bob Kenhard, Mike Wombwell, David Cheshire and the existing staff within Thermo VG Scientific involved in the management, development, sales and manufacturing of the Polaron Range.
We will continue to support the many Polaron instruments out in the field (including EM preparation instruments branded Polaron Equipment Ltd, Emscope, Bio-Rad Microscience, Fisons Instruments and VG). Additional details are available on the new Quorum WWW site ( http://www.quorumtech.com).
The new contact address (effective Nov. 12) will be
Quorum Technologies Unit 15A Euro Business Park New Road Newhaven East Sussex U.K. BN9 0DQ
Contacts: Mike.wombwell - sales (mike.wombwell-at-vgscientific.com - after 12th November - mike.wombwell-at-quorumtech.com) David Cheshire - technical support (david.cheshire-at-vgscientific.com - after 12th November - dave.cheshire-at-quorumtech.com) Bob Kenhard - sales and other enquiries (bob.kenhard-at-vgscientific.com - after 12th November - bob.kenhard-at-quorumtech.com) WWW: http://www.quorumtech.com US Contact: Energy Beam Sciences (telephone: 413 786 9322 E-mail - sales-at-ebsciences.com)
Mike Wombwell Polaron Range Sales Director Quorum Technologies
At 14:32 2001-11-06 -0600, Louis Somlai wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
Louis Somlai wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All, } } I think someone had once posted a URL to a website with an HTML } presentation that moved the observer from the sub-atomic viewing level to } one outside of our solar system using metric increments. Could someone } please send me that URL again? } } Thanks, } } Louis } Louis.Somlai-at-usm.edu
Would anyone know how to go about getting official certification for histo in the KS and MO region.
Thanks Tim Quinn University of Kansas Research Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
With effect from 1st November 2001 the Polaron Range of EM preparation instruments will change ownership from Thermo VG Scientific to Quorum Technologies Ltd.
Quorum Technologies Ltd. is the company formed by Bob Kenhard, Mike Wombwell, David Cheshire and the existing staff within Thermo VG Scientific involved in the management, development, sales and manufacturing of the Polaron Range.
We will continue to support the many Polaron instruments out in the field (including EM preparation instruments branded Polaron Equipment Ltd, Emscope, Bio-Rad Microscience, Fisons Instruments and VG). Additional details are available on the new Quorum WWW site ( http://www.quorumtech.com).
The new contact address (effective Nov. 12) will be
Quorum Technologies Unit 15A Euro Business Park New Road Newhaven East Sussex U.K. BN9 0DQ
Contacts: Mike.wombwell - sales (mike.wombwell-at-vgscientific.com - after 12th November - mike.wombwell-at-quorumtech.com) David Cheshire - technical support (david.cheshire-at-vgscientific.com - after 12th November - dave.cheshire-at-quorumtech.com) Bob Kenhard - sales and other enquiries (bob.kenhard-at-vgscientific.com - after 12th November - bob.kenhard-at-quorumtech.com) WWW: http://www.quorumtech.com US Contact: Energy Beam Sciences (telephone: 413 786 9322 E-mail - sales-at-ebsciences.com)
Mike Wombwell Polaron Range Sales Director Quorum Technologies
At the risk of looking very stupid, I will take your challenge.
Am I correct in understanding that your question has two parts? 1. What is the mean number of mitochondria per yeast cell in a population. 2. What is the mean area of cristae membranes.
If this is so, then there are actually three challenges here because to get an absolute number for the cristae surface area and the number of mitochondria per cell we also need to have an estimate for a reference space in actual numbers. A good reference space to estimate is cell volume.
Obtaining mean number of mitochondria per cell is actually very easy. I would first throw away the images already obtained because they were subjectively sampled.
We must then remove bias that may be a result of centrifugation - heavier (more dense) cells will pellet to the bottom, with the possibility that a gradient is formed in the pellet. To avoid this, embed the cells first in fast-setting gelatin or agarose, being careful to make sure the cells have been thoroughly mixed. We could of course do it properly and apply the "orintator" to make sure our sampling is isotropic (Mattfeldt et al 1989 Acta Stereol. 8:671-676).
Fix the block and cut it into slabs, rods and then cubes. Using a sequential random sampling protocol, take every "nth" block (decided using random number generator) and embed them separately in resin (see Lucucq 1993 Trends Cell Biol 3:345-358 for details).
Section for counting. A good way to sample the cells to count particles (here is where the catch comes in) is to only count the tops of the particles. With the yeast mitochondria, the 2-D profile does not always represent an individual particle. A profile may represent one single mitochondrion, or may represent a multiply branched, single mitochondrion, depending on the level of branching that is occurring. By counting the tops, connected profiles are eliminated from the sampled population. (for details on a possible method for estimating the amount of branching that is present, look for "Star Volume" in Gundersen et al 1986 J. Microsc. 143:3-45)
How to sample? Use the "Disector" (in this instance, the word means "2 sectors"). The reference is D.C. Stereo (a pseudonym for HJG Gundersen) 1984. J. Microsc. 134:127-136.
To apply, cut two sequential sections and take similar fields of view from both. Use one field to examine for mitochondrial profiles and the other to check if the profile is also present there ( the look-up frame). If a profile is present in one field but not present in the other, it is counted as a "top". (It is a variation of the 2-D counting rule, the associated tangent rule).
Knowing the area of the field of view and the thickness separating the two sequential sections, we can work out the volume being sampled. This will give us an estimate of the number of mitochondria per unit volume. This is not the number of mitochondria per cell. We still need to estimate the cell volume.
To estimate surface area, we can use simple cross lattice overlays and count the number of times a membrane profile crosses a test line. From this we can estimate length of membrane, and by knowing section thickness, can obtain an estimate of membrane area. (see H.J.G. Gundersen et al 1986 J. Microsc. 143:3-45. 1988 APMIS 96:379-394 & 857-881, T.M. Mayhew 1991 Exp. Physiol. 76:639-663, J. Lucocq for details on how to do this) We still need to estimate the cell volume.
How am I doing so far, Fred?
Now, a question for you. I can provide estimates of relative values for number and surface area. However, I do need a reference space to obtain absolute values. How would I obtain a reliable estimation of the mean cell volume of the yeast population? Presumably they are a rapidly dividing population.
Regards
Paul Webster.
on 11/7/01 12:53 PM, Monson, Frederick C. at fmonson-at-wcupa.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hypothetical! } } A graduate student in microbiology has just finished her first course in } electron microscopy, and she informs her advisor that she wants to study } mitochondrial structure and function in several related strains of yeast, } for one of which she fortuitously has electron micrographs of mitochondrial } profiles that are, she has been told, quite publishable. } } Her advisor suggests that she start by comparing 3-4 strains with respect to } number of mitochondria per cell and total mitochondrial cristal area under } identical conditions of culture. } } The student happily responds that she already has micrographs for regularly } sampled sections from serial sections she learned to make of the strain she } used in her EM course. She returns the next week with estimates of cristal } area and numbers of mitochondria per cell based on morphometric analysis of } those regularly sampled sections. } } If the cell is 12-25um in length (haploid and diploid) and always between } 3-5um in diameter, and if the mitochondrial profiles are in the range of } 1-2um (circular and ovoid, and sometimes or often branched), what should } the sampling criteria be for cells that are serially sectioned with sections } approximately 90nm in thickness? } } How could one derive the same, but apparently much more statistically } satisfactory, data from the 40+ cell profiles per section of cells randomly } oriented and not necessarily (or importantly) sectioned or analyzed } completely. } } All who take on this task should know there is an extremely important, and } possibly significant, hint that can be observed at: } http://www.msg.ucsf.edu/sedat/marsh/mito.html. I sometimes think it is well } to know of such hints (or at least to be aware of their possibility) before } attempting a task. Assumptions are almost always incorrect unless } rigorously defined and applied, and very often difficult to prove as such or } to isolate in generating an experimental design without flaws. } } Knowing that reality often gets in the way of statistics, and vice versa, I } am } } Yours most respectfully, because I wrestle with this problem every once in a } while and I haven't yet determined a satisfactory answer. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } CASI Home Page: http://darwin.wcupa.edu/casi/ } South Church Street } West Chester, PA, 19383 } Office: SS024 } Phone: 610-738-0437 } FAX: 610-436-3036 } eMail: fmonson-at-wcupa.edu } Please call before visiting } }
I am looking for a transformer to go into the Ion getter pump powersupply for a Philips 400T. The power supply transformer in this microscope is fried and needs to be replaced. I'm not sure who to contact since the microscope division of Philips is now owned by FEI. Any information would be appreciated.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have a Biorad MRC-1000 (1994) with the original video card, and do not have a service contract. The video signal goes into an Indec Systems box with an output RGB + horiz. + vert. via BNC connectors to the monitor. Lately we have been having sporadic severe problems where both the frame and image do not appear on the monitor. Instead a series of dots and/or lines moving across the monitor screen appear. This problem may persist for days and then disappear and the normal frame and image reappear. Sometimes the problem appears momentarily and then corrects. Jiggling the cables during the malfunction changes the rate of dots and lines, but does not bring the frame or image back. Most experts have said this is a monitor problem, but some suggest the video card. The monitor itself is specialized in that ordinary BNC monitors will not work satisfactorily (we have tried). A new monitor from Biorad is several thousand dollars. We would hate to buy a new monitor only to discover it really is the card. Any suggestions as to what additional troubleshooting can be done, or other insights, would be appreciated. Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id TAA24931 for dist-Microscopy; Thu, 8 Nov 2001 19:25:10 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id TAA24924 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 8 Nov 2001 19:24:39 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id TAA24916 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 8 Nov 2001 19:24:28 -0600 (CST) Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {p05100303b810df6617d6-at-[206.69.208.21]}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reimar_gaertner-at-wsib.on.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, November 8, 2001 at 12:19:54 ---------------------------------------------------------------------------
Question: My 8-year-old daughter and I have developed an interest in microscopy as a hobby. We have a Meade 9400 student microscope with built-in light (no field diaphram) and a fixed condenser (with a rotating aperture wheel). I am able to "focus" the condenser by inserting blocks under the slide (and lowering the stage to focus). This has allowed me to get closer to the critical focusing position (the surface of the light source now focuses about 1 cm above the stage. I need this for an effective dark-field effect at 100x magnification (10x10). I have heard that in order to best see thick specimens (especially at 400x), we need Kohler illumination. Do you think this will really make a difference? If so, what modifications would I need to make to achieve this? Many thanks. Reimar
Hi Listers, I am looking for independent providers of service contracts for a JSM-5310 SEM with Digital Image Grabber in Southern Arizona. I would appreciate it if anyone who could provide this service, or anyone who has experiences, good or bad, would email me off list.
on 11/8/01 3:48 PM, Paul Webster at pwebster-at-hei.org wrote:
} } At the risk of looking very stupid, I will take your challenge. } } Am I correct in understanding that your question has two parts? 1. What is } the mean number of mitochondria per yeast cell in a population. 2. What is } the mean area of cristae membranes. } } If this is so, then there are actually three challenges here because to get } an absolute number for the cristae surface area and the number of } mitochondria per cell we also need to have an estimate for a reference space } in actual numbers. A good reference space to estimate is cell volume. [skip] } } Regards
} Paul Webster.
Dear Paul, The risk of your looking stupid is very minimal. I'd use a mito-specific dye and LM to count mitos/cell (and get the distribution of cell sizes, info on whether [mitos] is greater for smaller/larger cells or is constant, etc.). I also have a bias for using HVEM tomography on thick sections to determine cristal-area/mito. Otherwise, yours seems like a very good plan. Yours, Bill Tivol
on 11/8/01 1:03 PM, Dusevich, Vladimir at DusevichV-at-umkc.edu wrote:
} } I am looking for publications of cell cultures observation in a ESEM. So } far not a great success. Any help is greatly appreciated. } Dear Vladimir, I don't know about ESEM, but I did look at cultured PTK1 cells in an environmental chamber in the HVEM (a TEM). I presented a few images at a Biophysical Society meeting about 15 years ago, so I do not remember many details. I'd be happy to discuss this with you off line if you want. Yours, Bill Tivol
Following is an announcement of a position opening at Washington University in St. Louis. I would appreciate your bringing this to the attention of anyone who might be interested. The workhorses of our facilities are a JEOL733 electron microprobe with Advanced Microbeam automation and image analysis, and a Rigaku Geigerflex X-ray diffractometer. Please respond to {blj-at-levee.wustl.edu} if you have any questions and not to the listserver.
Thanks!
Brad Jolliff _________
Research Specialist/Electron Probe Microanalysis
Washington University in St. Louis is seeking a research scientist who will operate and enhance the Electron Microprobe and X-ray Diffractometer facilities and conduct independent and collaborative research in petrology and geochemistry in the Department of Earth and Planetary Sciences. This is a Research Specialist position and is not tenure-track. Duties will include day-to-day management and operation of the EMP and XRD laboratories as well as enhancement of systems and analytical capabilities. Ph.D. preferred. For his or her research, the Specialist would have access to the wide array of analytical facilities within the Department of Earth and Planetary Sciences and in the McDonnell Center for the Space Sciences. These facilities include microbeam techniques (SIMS, laser-ablation ICP-MS, Raman spectroscopy), major- and trace-element chemical analysis (INAA, XRF, ICP-MS), mass spectrometers for stable and radiogenic isotope analysis, and a variety of other spectrometers including visible, IR, and Moessbauer.
To apply, submit a resume, statement of research interests, and names and contact information of three professional references to:
Bradley L. Jolliff Department of Earth and Planetary Sciences Washington University Campus Box 1169 One Brookings Drive St. Louis, MO 63130
Applications should be submitted by Dec 21, 2001. Women and underrepresented minorities are encouraged to apply. Washington University is an equal opportunity/affirmative action employer. Employment eligibility verification is required upon employment. __________
department web site: {http://epsc.wustl.edu/} __________
Bradley L. Jolliff, Ph.D Research Associate Professor Dept. of Earth & Planetary Sciences Campus Box 1169 Washington University One Brookings Drive St. Louis, MO 63130
I know you are looking for an independent service provider. However, in the past, I had been very pleased with JEOL service. Currently, I am working on a Hitachi EM, but for 12 years prior I worked on JEOL scopes (both TEM and SEM). The lab that I worked in was on the east coast in New Jersey. I'm not certain how the service is on the west coast, but the number one reason why I purchased new JEOL EMs was for their service history. We had an old JSM-35 and wanted to replaced it as well as purchase a new TEM. Based on the service we had received in the past, we immediately considered JEOL's line of EMs. Granted, this was about 7 years ago since the purchase and about 2 years since I've been out of that particular lab; however I was very pleased with the service from the north east division of JEOL. Good Luck with your search.
I intended on writing you right away, but I got too busy in the lab. I have tried this procedure a few times in the past with some success. I have two articles which I can fax to you if you would like. Sorry, I do not have an electronic copy. The references are:
Gonzalez-Angulo A, Ruiz De Chavez I, Castaneda M: A Reliable Method for Electron Microscopic Examination of Specific Areas from Paraffin-embedded Tissue Mounted on Glass Slides. American Society of Clinical Pathologists (1978) 697-700.
Bretschneider A, Burns W, Morrison A: "Pop-off" Technic. The Ultrastucture of Parafin-embedded Sections. American Journal of Clinical Pathology. Vol. 76, No. 4, October 1981. 450-453.
Jackie Garfield Reseach & Development Lifecell Corporation One Millennium Way Branchburg, NJ 08876 (908) 947-1182 jgarfield-at-lifecell.com
Pardon the off-topic post, but this is the only list to which I subscribe which has a significant number of members in Australia and East Asia, where the meteor shower has potential to be a once or twice in a lifetime experience this year:
For North America, best watching period is 4AM to 6AM EST on Nov. 18. For Australia and East Asia, predicted maximum is in the hours between midnight and about 4AM, depending on location, with possible maximum ZHR (zenithal hourly rates) approaching 10,000 per hour from dark-sky sites.
Astronomy is just like microscopy except we look at bigger things with bigger instruments, right?
thurston e herricksthurst0n-at-u.washington.edu11/8/01 3:59 PM
Thurston,
FEI is located in Hillsboro, OR.
Contact them at: 1-503-640-7500.
Good luck!
Gib
} Hello, } } I am looking for a transformer to go into the Ion getter pump powersupply } for a Philips 400T. The power supply transformer in this microscope is } fried and needs to be replaced. I'm not sure who to contact since the } microscope division of Philips is now owned by FEI. Any information would be } appreciated. } } Take care } } Thurston Herricks -- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
I have been asked to prepare some samples of brown adipose tissue for TEM, to look at mitochondrial structure and density. Has anyone got any suggestions for fixative and processing schedule for this type of material.
thanks
Kevin Electron Microscope unit Department of Zoology University of Aberdeen Aberdeen AB24 2TZ
Tel 01224-272847 Fax 01224-272396 ------------ Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
} Question: My 8-year-old daughter and I have developed an interest in } microscopy as a hobby. } We have a Meade 9400 student microscope with built-in light (no field } diaphram) and a fixed condenser (with a rotating aperture wheel). } I am able to "focus" the condenser by inserting blocks under the } slide (and lowering the stage to focus). } This has allowed me to get closer to the critical focusing position } (the surface of the light source now focuses about 1 cm above the } stage. } I need this for an effective dark-field effect at 100x magnification } (10x10). I have heard that in order to best see thick specimens } (especially at 400x), we need Kohler illumination. } Do you think this will really make a difference? If so, what } modifications would I need to make to achieve this? } Many thanks. } --------------------------------------------------------------------------- Reimar -
You'll find several suggestions for modifying your scope in Stevens' "The Microscope on a Budget"; here's the description from the MICRO bibliography (URL below):
Stevens, M.B. 1993 The Microscope on a Budget 247pp, 5.5x8.5", paperback, $13.00. ISBN 0-9638839-1-7 Logical Image Research, P.O.Box 1523, Buda, TX 78610. Also available from Nasco, 4825 Stoddard Rd., Modesto, CA 95356-9318 as #SB23799M; 800-558-9595. Subtitled "a complete guide to the low cost light microscope for the laboratory, photographers, and hobbyists", this book delivers solid information; it's a good buy. Homemade equipment (including exciting but seldom-discussed things like darkfield and Rhineberg illumination) is described in detail for the low-budget teacher or (adult-supervised) student. Both basic biological microtechnique and microcrystal preparation are described in detail. There are photography, supply source, reference, glossary, and index sections. High school - adult. RECOMMENDED
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Just wanted to send out a reminder about the ballot and M&M Journal subscription deadlines.
Ballots MUST BE postmarked by December 1, 2001 to be counted. M&M Journal subscription forms MUST BE returned by December 15, 2001 for the 2002 subscriptions.
Both the ballots and the subscription forms can be returned with your MAS renewal application.
If you have any questions, please contact me at rosslm-at-missouri.edu or (573) 882-4777 or at the the MAS email address or phone number listed below.
Thanks, Lou Ross MAS Membership Services masmembership-at-excite.com 1-800-4MASMEM (1-800-462-7636) www.microanalysis.org
on 11/9/01 11:01 AM, Rick Mott at rickmott-at-alumni.princeton.edu wrote: } } Astronomy is just like microscopy except we look at } bigger things with bigger instruments, right? } Dear Rick, That, and the fact that no one has yet invented electron astronomy (although study of the solar wind could be called proton astronomy)--let alone scanning probe astronomy (scanning black-hole force astronomy anyone?). :-) Yours, Bill Tivol
Counting the number of simple convex 3-D particles from some 3-D reference space (i.e. cell) using 2-D sections is not a trivial task. The sectioning process reduces the dimension of the particles to 2-D in the section. This 2-D sample of a particle is usually referred to as a profile. Many people have counted the number of profiles in a section and used this number to estimate the number of particles in the 3-D reference space. This of course is usually not a very good estimate of the TRUTH since the number of profiles in the 2-D sample is related not only to the number of particles but also the volume of the particles (more correctly the length of the particle perpendicular to the section plane) in the 3-D reference space. As Paul Webster pointed out in his earlier submission, Hans Gundersen solved the problem of counting particles in sections in his DISECTOR paper (J Microscopy 134:127-136, 1984).
Determining the number of complex particles, such as branching mitochondria adds complexity to the counting task. Luckily the hypothetical graduate student in Fred Monson's example did not take her EM class last year since Gundersen only solved the problem of easily counting complex particles using 2-D sections in September 2001 (J Microscopy 203:314-320). In this paper he introduces the ConnEulor principle for counting complex particles. Coincidentally the example he uses to demonstrate the ConnEulor principle is to determine the number of mitochondria in a cell. This technique like the original disector technique gives a density measure (number/volume) and you must still multiply by the average cell volume to obtain your estimate of particle number per cell.
A practical grad student might wonder if number of mitochondria is necessary and decide to measure the total volume of mitochondria within the cell (a much easier parameter to measure).
John
} } Hi Fred, } } At the risk of looking very stupid, I will take your challenge. } } Am I correct in understanding that your question has two parts? 1. What is } the mean number of mitochondria per yeast cell in a population. 2. What is } the mean area of cristae membranes. } } If this is so, then there are actually three challenges here because to get } an absolute number for the cristae surface area and the number of } mitochondria per cell we also need to have an estimate for a reference space } in actual numbers. A good reference space to estimate is cell volume. } } Obtaining mean number of mitochondria per cell is actually very easy. I } would first throw away the images already obtained because they were } subjectively sampled. } } We must then remove bias that may be a result of centrifugation - heavier } (more dense) cells will pellet to the bottom, with the possibility that a } gradient is formed in the pellet. To avoid this, embed the cells first in } fast-setting gelatin or agarose, being careful to make sure the cells have } been thoroughly mixed. We could of course do it properly and apply the } "orintator" to make sure our sampling is isotropic (Mattfeldt et al 1989 } Acta Stereol. 8:671-676). } } Fix the block and cut it into slabs, rods and then cubes. Using a } sequential random sampling protocol, take every "nth" block (decided using } random number generator) and embed them separately in resin (see Lucucq 1993 } Trends Cell Biol 3:345-358 for details). } } Section for counting. A good way to sample the cells to count particles } (here is where the catch comes in) is to only count the tops of the } particles. With the yeast mitochondria, the 2-D profile does not always } represent an individual particle. A profile may represent one single } mitochondrion, or may represent a multiply branched, single mitochondrion, } depending on the level of branching that is occurring. By counting the } tops, connected profiles are eliminated from the sampled population. (for } details on a possible method for estimating the amount of branching that is } present, look for "Star Volume" in Gundersen et al 1986 J. Microsc. } 143:3-45) } } How to sample? Use the "Disector" (in this instance, the word means "2 } sectors"). The reference is D.C. Stereo (a pseudonym for HJG Gundersen) } 1984. J. Microsc. 134:127-136. } } To apply, cut two sequential sections and take similar fields of view from } both. Use one field to examine for mitochondrial profiles and the other to } check if the profile is also present there ( the look-up frame). If a } profile is present in one field but not present in the other, it is counted } as a "top". (It is a variation of the 2-D counting rule, the associated } tangent rule). } } Knowing the area of the field of view and the thickness separating the two } sequential sections, we can work out the volume being sampled. This will } give us an estimate of the number of mitochondria per unit volume. This is } not the number of mitochondria per cell. We still need to estimate the cell } volume. } } To estimate surface area, we can use simple cross lattice overlays and count } the number of times a membrane profile crosses a test line. From this we } can estimate length of membrane, and by knowing section thickness, can } obtain an estimate of membrane area. (see H.J.G. Gundersen et al 1986 J. } Microsc. 143:3-45. 1988 APMIS 96:379-394 & 857-881, T.M. Mayhew 1991 Exp. } Physiol. 76:639-663, J. Lucocq for details on how to do this) We still need } to estimate the cell volume. } } How am I doing so far, Fred? } } Now, a question for you. I can provide estimates of relative values for } number and surface area. However, I do need a reference space to obtain } absolute values. How would I obtain a reliable estimation of the mean cell } volume of the yeast population? Presumably they are a rapidly dividing } population. } } Regards } } Paul Webster. } } } } } } } on 11/7/01 12:53 PM, Monson, Frederick C. at fmonson-at-wcupa.edu wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hypothetical! } } } } A graduate student in microbiology has just finished her first course in } } electron microscopy, and she informs her advisor that she wants to study } } mitochondrial structure and function in several related strains of yeast, } } for one of which she fortuitously has electron micrographs of mitochondrial } } profiles that are, she has been told, quite publishable. } } } } Her advisor suggests that she start by comparing 3-4 strains with respect } } to } } number of mitochondria per cell and total mitochondrial cristal area under } } identical conditions of culture. } } } } The student happily responds that she already has micrographs for regularly } } sampled sections from serial sections she learned to make of the strain she } } used in her EM course. She returns the next week with estimates of cristal } } area and numbers of mitochondria per cell based on morphometric analysis of } } those regularly sampled sections. } } } } If the cell is 12-25um in length (haploid and diploid) and always between } } 3-5um in diameter, and if the mitochondrial profiles are in the range of } } 1-2um (circular and ovoid, and sometimes or often branched), what should } } the sampling criteria be for cells that are serially sectioned with } } sections } } approximately 90nm in thickness? } } } } How could one derive the same, but apparently much more statistically } } satisfactory, data from the 40+ cell profiles per section of cells randomly } } oriented and not necessarily (or importantly) sectioned or analyzed } } completely. } } } } All who take on this task should know there is an extremely important, and } } possibly significant, hint that can be observed at: } } http://www.msg.ucsf.edu/sedat/marsh/mito.html. I sometimes think it is } } well } } to know of such hints (or at least to be aware of their possibility) before } } attempting a task. Assumptions are almost always incorrect unless } } rigorously defined and applied, and very often difficult to prove as such } } or } } to isolate in generating an experimental design without flaws. } } } } Knowing that reality often gets in the way of statistics, and vice versa, I } } am } } } } Yours most respectfully, because I wrestle with this problem every once in } } a } } while and I haven't yet determined a satisfactory answer. } } } } Fred Monson } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging(CASI) } } West Chester University of Pennsylvania } } Schmucker Science Center II } } CASI Home Page: http://darwin.wcupa.edu/casi/ } } South Church Street } } West Chester, PA, 19383 } } Office: SS024 } } Phone: 610-738-0437 } } FAX: 610-436-3036 } } eMail: fmonson-at-wcupa.edu } } Please call before visiting } } } } } } } } .
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
the much anticipated Leonid meteor show next Monday morning (Australian time) promises to be a spectacular event if the various predictions are even half correct. Unfortunately there are no guarantees (unlike eclipse predictions) so those of us making travel plans to observe the Leonids risk being in the wrong place at the wrong time. Personally I think the risk is worth taking considering the spectacular sight of thousands of meteors per hour if things work out.
In 1999 I drove for 5 hrs west of Sydney with a friend and my son in an attempt to observe a predicted reasonable Leonid shower. As it turned out we were treated to a much closer light show. The most ferocious thunder storm for twenty years devastate the local orchards in the area we chose to observe from and chased us all the way back to Sydney. We did manage to see a few dozen bright meteors through patchy clouds early in the night so it wasn't a total loss astronomically speaking.
This year I will be observing from central Australia, just a few kilometers from Ayres Rock. I figure that even if the sky show is clouded out or the predictions don't hold up, at least I can get a few good bush walks done during the days.
I'll give a brief report next week. Cheers,
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} ---------- } From: Rick Mott } Sent: Saturday, November 10, 2001 3:01 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: OT: possible spectacular Leonid meteors, Nov 18/19 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Pardon the off-topic post, but this is the only } list to which I subscribe which has a significant } number of members in Australia and East Asia, } where the meteor shower has potential to be } a once or twice in a lifetime experience this } year: } } } } } http://science.nasa.gov/headlines/y2001/ast08nov_1.htm?list534615 } } } } For North America, best watching period is 4AM } to 6AM EST on Nov. 18. For Australia and East } Asia, predicted maximum is in the hours between } midnight and about 4AM, depending on location, } with possible maximum ZHR (zenithal hourly rates) } approaching 10,000 per hour from dark-sky sites. } } Astronomy is just like microscopy except we look at } bigger things with bigger instruments, right? } } Rick Mott } }
-----Original Message----- } From: reimar_gaertner-at-wsib.on.ca [mailto:reimar_gaertner-at-wsib.on.ca] Sent: Thursday, November 08, 2001 8:19 PM Hi Reimar, Here's a cheap substitute for a Kohler type illumination I learned early in my microscopy career. Put a piece of translucent tape (magic mending type) on the rear of the slide. While this is not Kohler it comes close as it mimics the effect of light emitted close to the sample. The thinner the slide the better e.g. a cover slip. Good luck and have fun. Russ Gillmeister Xerox ~~~~~~~~~~~~~
Question: My 8-year-old daughter and I have developed an interest in microscopy as a hobby. We have a Meade 9400 student microscope with built-in light (no field diaphram) and a fixed condenser (with a rotating aperture wheel). I am able to "focus" the condenser by inserting blocks under the slide (and lowering the stage to focus). This has allowed me to get closer to the critical focusing position (the surface of the light source now focuses about 1 cm above the stage. I need this for an effective dark-field effect at 100x magnification (10x10). I have heard that in order to best see thick specimens (especially at 400x), we need Kohler illumination. Do you think this will really make a difference? If so, what modifications would I need to make to achieve this? Many thanks. Reimar
Unfortunately, we used our old JSM-35 as a trade-in. The purchase was about 7 years ago. The scope was in good shape, so I am sure it got a good home. However, you can check with JEOL for any used scopes. With a little luck, they just might have something. Also, you may want to check with Dick Daniel from Radco, Inc. (201) 891-8647. He services quite a bit of SEM equipment (mostly Denton products), and has many contacts throughout the United States. Perhaps, he knows of a good used SEM. It is worth a call. Good Luck,
Nanotubes and Peapods - Synthesis, Processing, Structure, Properties
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The experimental program involves electron microscopy including in-situ experiments, chemical synthesis and processing, and physical property measurements. The successful candidate should work well in a group, but be interested in developing and driving an aggressive, independent research effort.
The positions include health benefits and a competitive salary. We follow the University of Pennsylvania equal-opportunity guidelines and policies. If you would like to apply, please send a CV and the contact information of your references to David Luzzi at
Be aware, however, that most tape is anisotropic, subject to strain polarization, and will have a slight polarizing effect on transmitted light, perhaps preventing one from obtaining complete extinction between crossed polars or otherwise modifying expected optical effects.
John Twilley Conservation Scientist
Gillmeister, Russ wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } -----Original Message----- } } } From: reimar_gaertner-at-wsib.on.ca [mailto:reimar_gaertner-at-wsib.on.ca] } } Sent: Thursday, November 08, 2001 8:19 PM } Hi Reimar, Here's a cheap substitute for a Kohler type illumination I } learned early in my microscopy career. Put a piece of translucent tape } (magic mending type) on the rear of the slide. While this is not Kohler it } comes close as it mimics the effect of light emitted close to the sample. } The thinner the slide the better e.g. a cover slip. Good luck and have fun. } Russ Gillmeister } Xerox } ~~~~~~~~~~~~~ } } } Email: reimar_gaertner-at-wsib.on.ca } Name: Reimar Gaertner } } Organization: WSIB } } Education: Graduate College } } Location: Toronto, Ontario, Canada } } Question: My 8-year-old daughter and I have developed an interest in } microscopy as a hobby. } We have a Meade 9400 student microscope with built-in light (no field } diaphram) and a fixed condenser (with a rotating aperture wheel). } I am able to "focus" the condenser by inserting blocks under the } slide (and lowering the stage to focus). } This has allowed me to get closer to the critical focusing position } (the surface of the light source now focuses about 1 cm above the } stage. } I need this for an effective dark-field effect at 100x magnification } (10x10). I have heard that in order to best see thick specimens } (especially at 400x), we need Kohler illumination. } Do you think this will really make a difference? If so, what } modifications would I need to make to achieve this? } Many thanks. } Reimar } } --------------------------------------------------------------------------- } } }
In its trinocular head there sits a CF PL 5X Projection lens, above that is a Microflex HFX-IIA Photomicrographic Attachment, and onto that is a "35mm camera adapter A", then what is called in the HFX-IIA manual a "Motorized dark box FX-35WA". The latter looks more like a camera body to me.
We want to be able to mount our Nikon D 1 body onto the microscope, with or without the HFX-IIA, whatever it takes.
Is there anyone out there who can tell, from the above, what I need to do to acheive this?
I would have thought that all we need to do would be to mount the D 1 body directly onto the trinocular vertical tube, but then I know almost nothing about light microscopy.
tia
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Does anyone know who services or can tell me about how the mechanism for the LKB knifebreaker works? The knob that you turn to put pressure on the glass to break is very difficult to turn. Is there an easy way to clean or free it up? Will some solvent work or WD40? Or is there something else that I can do?
Thanks in advance.
Mei Lie -- Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
Dr Sims, first of all you need to get a C-Mount for the microscope, at the bottom of the trinocular head there are 3 hex-screw use them to remove the tube and insert the c-mount with your D1 camera, connect all the wires on the D1, and enjoy.
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My name is Kelly Stanard and I am a lab tech in the Image Analysis Laboratory at National Cancer Institute at Frederick, Maryland. I have been left with a Balzers freeze-fracture unit and some basic instructions but I have no idea what I am doing. Although a few of my attempts have produced decent replicas, I am very uncomfortable with the unit. What I am in need of are a few guidelines as to what all the buttons are for and when would be a good time to push them!! What I believe I have sitting here is a High Vacuum Coating Unit BAF 300 purchased in May, 1976. Quartz Crystal Thin Film Monitor QSG 201 Freeze Etching Unit Control BMS 101 Commutator Unit BCM 101 Pumping Unit Control DPA 101 Control Unit EVM 052A Microtom Movement Control BMB 101 Chamber IKR 010
If you can help me in anyway it would be greatly appreciated.
Thank you,
Kelly Stanard Image Analysis Laboratory NCI- Frederick P.O. Box B Frederick, MD 21702 301-846-1528 kstanard-at-ncifcrf.gov
Dr Sims, first of all you need to get a C-Mount for the microscope, at the bottom of the trinocular head there are 3 hex-screw use them to remove the tube and insert the c-mount with your D1 camera, connect all the wires on the D1, and enjoy.
Alfredo Hernandez
"Ritchie Sims" {r.sims-at-auckland.ac.nz} wrote:
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You can try calling Leica for service, however you may have to send the knife-maker out. I use a company called Dolbey-Jamison, located in PA. They do in-house service. Good Luck,
CRYOMICROSCOPY GROUP ANNUAL MEETING EM Centre, University of Birmingham, Birmingham, UK
Wednesday 21st November 2001
PROGRAMME :
9.30 - 10.30 Registration, Coffee and Trades Exhibition 10.30 Chairman's Welcome and Introduction 10.35 -11.15 Charge elimination for the X-ray microanalysis of frozen hydrated specimens Patrick Echlin, Cambridge Analytical Microscopy, Cambridge 11.15 -12.00 Freeze-substitution and 3D reconstruction of muscle Pradeep Luther, Imperial College, London 12.00 -12.45 Silver enhancement for immunolabelling Jan Leunissen, Aurion, Wageningen, The Netherlands 12.45 - 14.00 Lunch , Trades Exhibition & Posters 14.00 -14.15 Annual General Meeting 14.15 - 15.00 RMS Beginners Competition 15.00 -15.45 Ultrastructural localisation of Prion protein in the brain using cryo-immunogold electron microscopy Peter Peters, Netherlands Cancer Institute, Amsterdam 15.45 - 16.00 Problems with preparing articular cartilage for microscopic study Iolo ap Gwynn, S C Wade and R G Richards. University of Wales, Aberystwyth 16.00 Closing Remarks and Tea
FURTHER DETAILS : http://www.cryomicroscopygroup.org.uk/index.htm
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
l.tetley-at-bio.gla.ac.uk tel 0141 330 4431 FAX 0141 330 3516
Use acetonitrile instead of ethanol or acetone for dehydration. Also, use imidazole buffered osmium to postfix. The mitochondria come out beautifully. I have done this and it works. The references are:
Edwards, H. H. et al. 1992. Microscopy Research and Technique Volume 21, pp 39-50.
Angermuller, S. and H. D. Fahimi. 1982. Histochemical Journal Volume 14, pp. 823-835.
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Dear Listers, I am expecting the following samples for paraffin embedding sometime soon -- "wolf ovaries have been preserved in tequila for the past 6 months or so". I first assumed that this was a joke but apparently they are serious--this is a reproductive physiology project from an ecology lab. Any comments are welcome--levity included. I can hear your laughter already! My questions: --has anyone used tequila as a fixative? -- is the embedding worth pursuing? Rosemary Walsh
We service and sell reconditioned LKB Knife Makers. Please contact me off line. We are closed from Nov. 15 to 26. Regards, Microscopy Labs P.O.Box 338 61 West Street Red Bank, NJ 07701 732 727 6228 fax 732 758 9142
----- Original Message ----- } From: "Mei Lie Wong" {wong-at-msg.ucsf.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, November 12, 2001 8:43 PM
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No experience and can't think of any jokes but .....
I guess tequila is about 40% ethanol or is this home-brewed stuff?? Standardisation in the future could be a problem - will they be using the same brand next time?
Limited lipid extraction in that case - wash in PBS and cryosection?
You could rehydrate to PBS and refix in buffered formaldehyde - could help to protect against processing.
I guess that wolf ovaries are resonably large so you could try various combinations?
I would be interested to hear how it goes. Go for it and educate us all!
At 19:46 2001-11-13 -0500, Rosemary Walsh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
I haven't worked in biological EM for some time. I do, however, have some experience with the stated topic. One evening some years ago I partook a bit too much tequila and had a howling good time. Unfortunately, the next morning I found that I was very well fixed indeed. Needed the hair of the dog (or wolf as the case may be) to survive the day. That memory has been well embedded in my memory as I wax nostalgic.
I have no other comments aside from this nonsense. Good luck.
Gary M. Brown
Rosemary Walsh {rw9-at-psu.edu} To: Microscopy-at-sparc5.microscopy.com cc: Subject: a sample prep queri 11/13/01 06:46 PM
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Dear Listers, I am expecting the following samples for paraffin embedding sometime soon -- "wolf ovaries have been preserved in tequila for the past 6 months or so". I first assumed that this was a joke but apparently they are serious--this is a reproductive physiology project from an ecology lab. Any comments are welcome--levity included. I can hear your laughter already! My questions: --has anyone used tequila as a fixative? -- is the embedding worth pursuing? Rosemary Walsh
Dear listers, Some time ago one of you discussed a combination of resins that snapped off culture dishes fairly easily. Could you post that again as I need the information and can't seem to locate it in the archives. I've tried several combinations of resins from different sources on different brands of plates and the results are not great. There also appear to be differences in the "snapability" between the multiple well plates and the individual dishes from the same companies, which adds to the problem. I'd really appreciate knowing the magic combination of resin components if you could readdress the issue. Thanks, Mary Gail Engle
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
} --has anyone used tequila as a fixative? } -- is the embedding worth pursuing? } Rosemary Walsh ****************** Hi Rosemary, that's a new one.
My first question is, what proof was the tequila? If it was potent stuff, the ethanol level may have been high enough to act as a coagulative fixative. Many years ago I was asked to do EM of the gills of clams that had been fixed in 70% ethanol. I rehydrated them to buffer and then processed for EM as usual (glut, osmium, etc). The ultrastructure, while clearly compromised was far better than I had expected. You can try this with the ovary samples. The fact that they are destined for paraffin means that you won't have the very fine structure to worry about. You might want to use a higher than usual concentration of pfa, or even throw in a little glut to lock in whatever is left.
Good luck, Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I would like to contract out cryosectioning of some carbon black - filled rubber material. We desire sections on the order of 1 micron thick amenable to light microscope/image analysis measurements that we will perform. There is plenty of sample material and we don't need serial sectioning. We are hoping to have this done within weeks.
If you are interested or know of someone who may be, and/or have questions, please contact me off line at tgruber-at-phelpsd.com.
Not too dumb on the part of the enviro's. After all, Tequila does the worm! Tequila, as I quickly discovered, can have between 35-55% ethanol.
My recommendation would be to proceed with your own fixation and not worry. They used their heads when they realized they went into the field unprepared, or they had an accident, or they couldn't take the fixative with them and couldn't find any there. You treat the tissue as if it just got to you. The nitrogens are likely still waiting for the formaldehyde. Or, you could transfer them to Clarke's 3:1(Absolute ethanol:Glacial Acetic Acid), and have a really granular texture in the sections. Or Bouin's, which would also complete the job on the worm!
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Rosemary Walsh } Sent: Tuesday, November 13, 2001 7:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: a sample prep queri } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } I am expecting the following samples for paraffin embedding sometime } } soon -- "wolf ovaries have been preserved in tequila for the past 6 months } } or so". I first assumed that this was a joke but apparently they are } serious--this is a reproductive physiology project from an ecology } lab. Any comments are welcome--levity included. I can hear your laughter } } already! My questions: } --has anyone used tequila as a fixative? } -- is the embedding worth pursuing? } Rosemary Walsh } } }
As I recall the lower range is around that for normal cells and the upper range is something like 15-40um (BIG!!!). Just hope that J. Kiernan sees your email so that you get a more refined result.
Fred Monson
} ---------- } From: Cheri Owen } Sent: Tuesday, November 13, 2001 11:09 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Range of size of neuronal nuclei } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anyone know off the top of their head what the range of size of } neuronal nuclei is? } } Cheri Owen } Dept. Emergency Medicine/Physiology } Wayne State University } } }
Position Open Microscopy and Microanalysis Research Assistant
The NCSU Analytical Instrumentation Facility (AIF) is seeking qualified applicants for a SEM Microscopist position opening December 11, 2001.
Duties and responsibilities include: Operation and maintenance of SEM/EDS instrumentation and sample preparation and analysis equipment; scheduling of access to and oversight of the above instrumentation; user training and assistance; assistance with the teaching of electron microscopy laboratory classes, and analysis of a wide variety of samples. Qualifications must include a minimum of a BS with 5 years hands on SEM/EDS experience, MS with 2 years hands on SEM/EDS experience or a Ph.D. with 1 year hands on SEM/EDS experience. Experience must be in a non biological materials related discipline. Required skills include: extensive hands on experience with SEM and related techniques and accessories (e.g. EDS, specimen preparation and associated analytical tools). Preferred qualifications include: teaching and user training; familiarity with modern electronics; and experience with vacuum systems.
Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 318A EGRC; 1010 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity and Affirmative Action Employer. ADA Accommodations: Phil Russell, prussell-at-ncsu.edu, 919-515-7501. In its commitment to diversity and equity, North Carolina State University seeks applications from women, minorities, and persons with disabilities
It's possible, that the ovaries are fixed OK, provided they were cut open when placed into the tequila. Normally, 70% ethanol would have been better, maybe they should have used for alcohol fixation. If they have not been cut open prior to fixation, then the odds are that the morphology of the internal areas of the ovary will not be very good. However, try one sample, trim to desired sized and post-fix in 10% formalin overnight, process for paraffin embedding.
Good Luck!
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Rosemary Walsh [mailto:rw9-at-psu.edu] Sent: Tuesday, November 13, 2001 7:46 PM To: Microscopy-at-sparc5.microscopy.com
Dear Listers, I am expecting the following samples for paraffin embedding sometime
soon -- "wolf ovaries have been preserved in tequila for the past 6 months or so". I first assumed that this was a joke but apparently they are serious--this is a reproductive physiology project from an ecology lab. Any comments are welcome--levity included. I can hear your laughter already! My questions: --has anyone used tequila as a fixative? -- is the embedding worth pursuing? Rosemary Walsh
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
My name is Kelly Stanard and I am a lab tech in the Image Analysis Laboratory at National Cancer Institute at Frederick, Maryland. I have been left with a Balzers freeze-fracture unit and some basic instructions but I have no idea what I am doing. Although a few of my attempts have produced decent replicas, I am very uncomfortable with the unit. What I am in need of are a few guidelines as to what all the buttons are for and when would be a good time to push them!! What I believe I have sitting here is a High Vacuum Coating Unit BAF 300 purchased in May, 1976. Quartz Crystal Thin Film Monitor QSG 201 Freeze Etching Unit Control BMS 101 Commutator Unit BCM 101 Pumping Unit Control DPA 101 Control Unit EVM 052A Microtom Movement Control BMB 101 Chamber IKR 010
If you can help me in anyway it would be greatly appreciated.
Thank you,
Kelly Stanard Image Analysis Laboratory NCI- Frederick P.O. Box B Frederick, MD 21702 301-846-1528 kstanard-at-ncifcrf.gov
The CCD on the D1 is 23.7mm x 15.6mm, so a projection lens of 1.5x should be sufficient. Do you have some kind of C-mount to Nikon mount converter? The D1 uses the same Nikon "F" mount that Nikon SLR bodies have used for years, so without a converter it will not mount directly to a C-mount.
George
George Laing National Graphic Supply
The only question now is which C-mount. Whereas the CCD for Coolpix mounts requires 1X, the CCD for the D1 is larger ... 1.5X, 2X???
C-Mount is just an adapter and wont requiere any additional lens, You'll get a direct image. You can use different c-mounts with zoom in them in order to get a larger field of view. The lens you require will depend on the chip size of your camera (DN100 has 1/2" chip size) so if you want a larger field of view you probably want to buy a C mount + TV adapter 0.45X The DXM1200 has a 2/3" chip size therefore the best combination will be C mount + TV adapter 0.6X
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Hi folks, Thanks for the replies. Apparently this was serious inquiry. Samples were taken in a remote area in Alaska. The interest is in determining the number of ovulations compared with the number of pregnancies in dominant wolves versus pseudo pregnancies in other pack females. We'll probably rehydrate to PBS and fix in 4% paraformaldehyde, process and embedd in paraffin, stain with H&E for LM to see what we have. Another option is to go directly to 10% formalin. If I can obtain some sample I intend to process for TEM "just for the record". This should take us awhile but I promise to keep you posted. Thanks again to all of you wonderful folks who responded. Not yet destined for Margaritaville! Rosemary
Here are the replies No experience and can't think of any jokes but .....
I guess tequila is about 40% ethanol or is this home-brewed stuff?? Standardisation in the future could be a problem - will they be using the same brand next time?Limited lipid extraction in that case - wash in PBS and cryosection? You could rehydrate to PBS and refix in buffered formaldehyde - could help to protect against processing. I guess that wolf ovaries are resonably large so you could try various combinations? Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
Sheesh! What an ecology lab. Were they getting the wolves drunk and then ripping out their ovaries and plunking them in tequila? I've never tried preserving anything in tequila, but it probably works for livers. Good luck (and you should charge these idiots A LOT). Mary McKee MGH Renal Unit Charlestown, MA 02129 (617)726-3696
Hi Rosemary, After 6 months or so, do you suppose that the ovaries are "Wasting Away in Margaritaville"? JME Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA phone: 506-364-2519 fax: 506-364-2505 email: jehrman-at-mta.ca
In "The collection and preservation of Animal Parasites" by G.O.W. Kruse e M.H. Pritchard (Tech. Bull. no 1, The Harold W. Manter Laboratory- University of Nebraska Press, 1982) the use of any available local brew to fix and preserve parasites is reported, if no fixative is available.Transfer of parasites to 70% EtOH as soon as the latter is available, after carefully pouring out the spirit.Bruno Dore
I have been present at the AFIP when they got specimens similar to your wolf ovaries. Some of us wanted to go do field work with the guys who came up with this as they said they use tequila alot, for many things. Our imaginations went active. You might try contacting the Histology laboratory there and find out what they do exactly. I can give you some advise a fellow histologist and I will forward yourmessage to HistoNet as someone there may be able to assist you also. The first issue is the fixation, at this point both fixation and dehydration are complete. Do you plan to dissect the specimens into blocks, if so what sizes? I have a protocol using butanol to clear the specimen prior to infiltration with paraffin. It is very gentle and would hopefully allow the perimeters to stay softer and easier to section. Let me know if I can assist you or you need the protocol. I am generally in from 9AM to 5PM. Today will be the exception we have meetings until 1PM.
Pamela A. Marcum Histology/Microscopy Product Development Manager 400 Valley Road Warrington, PA 18976 Phone: 800-523-2575 Ext 167 215-343-6484 Ext 167 Fax: 215-343-0214 E-mail: pmarcum-at-polysciences.com
My first question is, what proof was the tequila? If it was potent stuff, the ethanol level may have been high enough to act as a coagulative fixative. Many years ago I was asked to do EM of the gills of clams that had been fixed in 70% ethanol. I rehydrated them to buffer and then processed for EM as usual (glut, osmium, etc). The ultrastructure, while clearly compromised was far better than I had expected. You can try this with the ovary samples. The fact that they are destined for paraffin means that you won't have the very fine structure to worry about. You might want to use a higher than usual concentration of pfa, or even throw in a little glut to lock in whatever is left. Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
For TEM I have looked at skin and underlying connective tissue from all kinds of strange scenarios, eg. collected from a dead emu under freezing weather conditions for a number of days. Expect loss of fine detail and for certain organelles to be much more poorly preserved than others. You might take some samples and run them down to buffer then refix in buffered formaldehyde. Embed some other samples straight from the tequila. I suspect the tequila is between 40-60% ethanol. Don't waste the tequila, maybe you can start a fad, a pleasent way to increase fertility, or some other patent medicine application. Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Laboratory University of Kansas
Not too dumb on the part of the enviro's. After all, Tequila does the worm! Tequila, as I quickly discovered, can have between 35-55% ethanol. My recommendation would be to proceed with your own fixation and not worry. They used their heads when they realized they went into the field unprepared, or they had an accident, or they couldn't take the fixative with them and couldn't find any there. You treat the tissue as if it just got to you. The nitrogens are likely still waiting for the formaldehyde. Or, you could transfer them to Clarke's 3:1(Absolute ethanol:Glacial Acetic Acid), and have a really granular texture in the sections. Or Bouin's, which would also complete the job on the worm.Regards,Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu
I do a lot of ovaries here - A LOT, but they are usually rodent and are fixed in 10% formalin, Bouins or 70% ETOH not tequila (what a waste of good tequila - assuming that it was of the good variety). I hope that the ovaries were small or trimmed before they were plunked into the tequila. I'd process them mostly out of curiosity to see how they might turn out. The ultrastructure wouldn't be much to speak of but there might be some histology to see. One reply on the microscopy listserver stated that tequila is about 40% alcohol. If so I'd trim the samples if necessary and process as I usually do by starting in 70% ETOH through 80%, 95% x2, 100% x3, Xylene x 2, paraffin x 2. The replier on microscopy also said to rehydrate in PBS and refix in formalin. You could do that. In fact this might be a good idea it might help to preserve what hasn't already been destroyed. I hope Rosemary processes the wolf ovaries and gives a report on the outcome.
Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : Sr. Research Specialist University of Arizona : (office: AHSC 4212) P.O. Box 245044 : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : (FAX: 520-626-2097) (email: algranth-at-u.arizona.edu) :
I think the active ingredient would be the ethanol content of tequila. No, I am not laughing, when Lord Byron died somewhere in Mediterrean area or whereever, his body was preserved in a wine or brandy cask and shipped home (to England?) for burial.
She might want to know what the percentage of alcohol is, what proof, and work on processing from that point. I would love to know the outcome of this, I used Chinese vodka as a means to clean a scratch in skin while in that country and prevent a skin infection, it worked but I had a quart of the stuff, gave it to some person who loved the stuff in coca cola! Skin healed well, his liver suffered badly!
As for humor, it could be said the tissues were well "pickled", same as people are when they ingest too much of this stuff!! Gayle Callis MT,HT,HTL(ASCP) Histopathology Supervisor Veterinary Molecular Biology - Marsh Lab Montana State University - Bozeman 19th and Lincoln St Bozeman MT 59717-3610
406 994-6367 406 994-4303 (FAX)
It's possible, that the ovaries are fixed OK, provided they were cut open when placed into the tequila. Normally, 70% ethanol would have been better, maybe they should have used for alcohol fixation. If they have not been cut open prior to fixation, then the odds are that the morphology of the internal areas of the ovary will not be very good. However, try one sample, trim to desired sized and post-fix in 10% formalin overnight, process for paraffin embedding. Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
The C-mount interface suggestions below should work and will result in an image being available to your D1 sensor. However, if your solution requires that you invest more money into the process, you might consider additional alternatives geared more towards 35mm SLR photography to optimize your results. With the D1 being an SLR camera body, and given the pixel resolution of the sensor, for our D1 customers, we have, instead, used one of two options which, we believe, result in optimal sensor fill and end results.
1.) Diagnostic Instruments has a photographic adapter series for coupling 35mm SLR cameras to microscopes. Within that series, the PA1-12A with the NIKC-T2 adapter usually fits the bill for most Nikon scopes. Diagnostic Instruments recommends a Nikon 1.6x photoeyepiece for image projection.
2.) Alternatively, I believe Nikon now has a direct Nikon option for the D1 series of cameras. It does pretty much the same thing as Option 1, providing the 1.6x projection, except it all comes from Nikon, and might be slightly more "refined".
Diagnostic Instruments products are available through a network of dealers, most of which are also microscope dealers. Nikon products are (obviously) available from your local Nikon dealer. You might consult your Nikon dealer in regard to either solution. They can probably offer both.
PLEASE NOTE: This e-mail is in no way meant to pass judgment on the suggestions of other list members. I only provide the above info based on opinion and the approach we have taken in this situation.
-----Original Message----- } From: "tartenon-at-netscape.net"-at-sparc5.microscopy.com [mailto:"tartenon-at-netscape.net"-at-sparc5.microscopy.com] Sent: Wednesday, November 14, 2001 4:08 PM To: Microscopy-at-sparc5.microscopy.com
C-Mount is just an adapter and wont requiere any additional lens, You'll get a direct image. You can use different c-mounts with zoom in them in order to get a larger field of view. The lens you require will depend on the chip size of your camera (DN100 has 1/2" chip size) so if you want a larger field of view you probably want to buy a C mount + TV adapter 0.45X The DXM1200 has a 2/3" chip size therefore the best combination will be C mount + TV adapter 0.6X
} } Dear listers, } Some time ago one of you discussed a combination of resins that } snapped off culture dishes fairly easily. Could you post that again } as I need the information and can't seem to locate it in the } archives. I've tried several combinations of resins from different } sources on different brands of plates and the results are not great. } There also appear to be differences in the "snapability" between the } multiple well plates and the individual dishes from the same } companies, which adds to the problem. I'd really appreciate knowing } the magic combination of resin components if you could readdress the } issue. } Thanks, } Mary Gail Engle } **************** HI Mary Gail,
I was the one who posted my "magic mixture". It is this: LX112 (Ladd Industries), DDSA (Electron Microscopy Sciences), NMA (also EMS) and DMP-30 (Ladd). I use a "standard" Epon recipe, nothing secret there. After the last 100% ethanol, I pour a shallow layer of resin into the dish and insert (tubes made by cutting the ends off BEEM capsules) into the resin over areas of interest. Be sure that the molded end of the tube is facing down so that you have a perfectly flat edge. Polymerize overnight, then fill just the tubes with more resin. Return to the oven for 24-36 hr. when you remove then dish from the oven, just grasp the embedding tube with a pair of needle-nosed pliers and snap. If anything, a small amount of the dish may come off with your block, but it shoul.d not leave any cells behind. I haven't seen a difference between single and multi-well plates, although the small diameter wells often don't work well....I think its tough to get a good exchange of the last alcohol and the blocks tend to be gummy.
If you have any more questions, contact me off-list and will will elaborate.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi listers, With the agave shortage a couple of years back, tequila is getting expensive. What about gin? Or whiskey? Then we could give the fixations cute names like 'The Limey' or 'Kentucky's Finest' (respectively). The ethanol content would be about the same, and there would be less impurities (unless those impurities are what makes it a good fixative). Just a thought.
Gareth Morgan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } No experience and can't think of any jokes but ..... } } I guess tequila is about 40% ethanol or is this home-brewed stuff?? } Standardisation in the future could be a problem - will they be using the } same brand next time? } } Limited lipid extraction in that case - wash in PBS and cryosection? } } You could rehydrate to PBS and refix in buffered formaldehyde - could help } to protect against processing. } } I guess that wolf ovaries are resonably large so you could try various } combinations? } } I would be interested to hear how it goes. Go for it and educate us all! } } At 19:46 2001-11-13 -0500, Rosemary Walsh wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } I am expecting the following samples for paraffin embedding sometime } } soon -- "wolf ovaries have been preserved in tequila for the past 6 months } } or so". I first assumed that this was a joke but apparently they are } } serious--this is a reproductive physiology project from an ecology } } lab. Any comments are welcome--levity included. I can hear your laughter } } already! My questions: } } --has anyone used tequila as a fixative? } } -- is the embedding worth pursuing? } } Rosemary Walsh } } } } } } Med vänliga hälsningar/With best wishes } } Gareth } } "Close your eyes and look. What you saw at first is there no more; and what } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-impi.ki.se } } Tel +46 8 728 3734 } Fax +46 8 728 3688 } } Gareth Morgan MPhil MSc FIBMS, } Institutionen för Mikrobiologi, } Patologi och Immunologi(IMPI), H5, } Karolinska Institutet, } Dept of Biomedical Laboratory Science, } Lindhagensgatan 92, Box 12773, } S 112 96, Stockholm } Sweden } } OBS! Besöksadress: Lindhagensgatan 92 } NB! Visiting address:Lindhagensgatan 92
-- Russell McConnell Confocal Imaging Facility Technician Department of Neuroscience Tufts University School of Medicine M&V Building room #137 136 Harrison Ave. Boston, MA 02111 Tel. (617) 636-3795
Hi It all depends on which measurements you have in mind. In the simplest case, you can use topographic imaging to trace the changes in topography during oxidation. From these measurements, you can detect the formation of oxide if it is accompanied by significant morphological changes, e.g. instead of flat surface with atomic steps you see large (10-100 nm) oxide crystallites. Of course, if oxide is uniform, you wouldn't see much. If you want to get oxide thickness, you can etch it locally and measure the step heigh between etched and unetched parts. Electrostatic force microscopy and surface potential microscopies are completely different story. In principle, on the potential map you can see charged grain boundaries, trapped charges and other electroactive defects. You can also measure CPD variations between different phases. Some useful leads can be found in papers by J.W.P. Hsu, D.A. Bonnell et al and Ludeke and Cartier, among others. However, interpretation of the SSPM images can be tricky. The usual caveat is topographical artifacts, etc. You might also consider asking the same question at DI forum at spm-at-di.com Hope this helps Sergei
} } Hi, I have a question about the GaSb. } } How to use AFM/EFM to characterize the oxidation layer which is above the } GaSb surface. } } Thank you for your help! } } Xianglin Li } } University of Massachusetts } 978-934-3411
-- Sergei V. Kalinin Dept. Mat. Sci. Eng. University of Pennsylvania, 3231 Walnut St, Philadelphia PA 19104 Tel: (215) 898-3446 Fax: (215) 573-2128 E-mail: sergei2-at-seas.upenn.edu URL: http://www.seas.upenn.edu/~sergei2
Bacardi 151 proof rum made a good fixative for marine organisms ...
Phil
} Hi listers, } With the agave shortage a couple of years back, tequila is getting } expensive. What about gin? Or whiskey? Then we could give the } fixations cute names like 'The Limey' or 'Kentucky's Finest' } (respectively). The ethanol content would be about the same, and there } would be less impurities (unless those impurities are what makes it a } good fixative). Just a thought. } } Gareth Morgan wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } No experience and can't think of any jokes but ..... } } } } I guess tequila is about 40% ethanol or is this home-brewed stuff?? } } Standardisation in the future could be a problem - will they be using the } } same brand next time? } } } } Limited lipid extraction in that case - wash in PBS and cryosection? } } } } You could rehydrate to PBS and refix in buffered formaldehyde - could help } } to protect against processing. } } } } I guess that wolf ovaries are resonably large so you could try various } } combinations? } } } } I would be interested to hear how it goes. Go for it and educate us all! } } } } At 19:46 2001-11-13 -0500, Rosemary Walsh wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Listers, } } } I am expecting the following samples for paraffin embedding sometime } } } soon -- "wolf ovaries have been preserved in tequila for the past 6 months } } } or so". I first assumed that this was a joke but apparently they are } } } serious--this is a reproductive physiology project from an ecology } } } lab. Any comments are welcome--levity included. I can hear your laughter } } } already! My questions: } } } --has anyone used tequila as a fixative? } } } -- is the embedding worth pursuing? } } } Rosemary Walsh } } } } } } } } } } Med vänliga hälsningar/With best wishes } } } } Gareth } } } } "Close your eyes and look. What you saw at first is there no more; and what } } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } } } http://www.ki.se/biomedlab } } e-mail Gareth.Morgan-at-impi.ki.se } } } } Tel +46 8 728 3734 } } Fax +46 8 728 3688 } } } } Gareth Morgan MPhil MSc FIBMS, } } Institutionen för Mikrobiologi, } } Patologi och Immunologi(IMPI), H5, } } Karolinska Institutet, } } Dept of Biomedical Laboratory Science, } } Lindhagensgatan 92, Box 12773, } } S 112 96, Stockholm } } Sweden } } } } OBS! Besöksadress: Lindhagensgatan 92 } } NB! Visiting address:Lindhagensgatan 92 } } -- } Russell McConnell } Confocal Imaging Facility Technician } Department of Neuroscience } Tufts University School of Medicine } M&V Building room #137 } 136 Harrison Ave. } Boston, MA 02111 } Tel. (617) 636-3795
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Microscopy technician vacancy in Albany, California (East San Francisco Bay Area), near Berkeley, California.
An individual is needed to provide assistance and expertise in scanning electron and light microscopy on various projects in food and non-food applied research. In addition, the person will provide expertise in photography, imaging, and preparation of graphics for presentation and publication using digital cameras and Windows-based computers. Applicants will be considered at the GS-5 through GS-7 levels.
Those interested, please see the Federal website for further information about the position and how to apply.
I am looking for a supplier for Barium permanganate (for use as a TEM Stain), and I can't find anyone who carries it (Fisher, Sigma, EMS, Pella, SPI, Ladd, etc.).
Any suggestions?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I have just seen an Imax film "Cosmic Journey" which expands on the plot of "Powers of Ten" adds some graphics and presents a (speeded up!) depiction of the early development of the Universe and all that. Its worth looking out for.
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
Hello, I have been loaned a microtome (Leitz 1512) by a colleague which according to the instruction manual requires regular oiling. The microtome did not come with any oil and the price I have been quoted by a local supplier for microtome oil was about 75$US (including VAT) for 50ml (SAKURA 1447). I found the price a bit steep and am left wondering if it is absolutely necessary to buy "microtome" oil or if I can use an off the shelf oil (e.g 3 in 1 or something like that). I do not want to compromise the performance of the microtome as it is not mine and would appreciate any advice from those with experience. Thanks for any help. Sincerely, Jonathan
Jonathan Wilson (PhD) CIIMAR Rua do Campo Alegre 823 4150-180 Porto, Portugal tel: 351 22 608 0470 / 71 fax: 351 22 606 0423 e-mail: wilson_jm-at-cimar.org web: www.cimar.org
Hello, I have been loaned a microtome (Leitz 1512) by a colleague which according to the instruction manual requires regular oiling. The microtome did not come with any oil and the price I have been quoted by a local supplier for microtome oil was about 75$US (including VAT) for 50ml (SAKURA 1447). I found the price a bit steep and am left wondering if it is absolutely necessary to buy "microtome" oil or if I can use an off the shelf oil (e.g 3 in 1 or something like that). I do not want to compromise the performance of the microtome as it is not mine and would appreciate any advice from those with experience. Thanks for any help. Sincerely, Jonathan
Jonathan Wilson (PhD) CIIMAR Rua do Campo Alegre 823 4150-180 Porto, Portugal tel: 351 22 608 0470 / 71 fax: 351 22 606 0423 e-mail: wilson_jm-at-cimar.org web: www.cimar.org
We are trying to microscopy a microorganism, spiroplasma using TEM. Every time we had problem to see clearly the membrane around the organism. We tried to increase the time of Osmium (4%) fixation, 3-5 hours at RT after glutaldehyde fixation. But the membrane preservation is still terrible. I would very much appreciate any of suggestions for that.
The Integrated Microscopy Core, Department of Molecular and Cellular Biology, Baylor College of Medicine has an immediate full-time opening for an electron microscopy technician. The Integrated Microscopy Core is a busy, state-of-the-art facility with a new Hitachi H7500 equipped with Gatan’s new 2Kx2K CCD camera, deconvolution, laser scanning confocal, and 2 CCD-based upright and inverted epifluorescence microscopes.
The applicant should have at least one year of experience in various aspects of sample preparation for biological TEM. This should include fixation, embedding, ultrathin sectioning staining and darkroom procedures, TEM operation and knowledge of computers. Other duties include preparation of solutions, embedding media and excellent organizational skills.
The position offers opportunities for training in advanced microscopy techniques, including digital TEM, fluorescence, deconvolution and laser scanning confocal microscopy. Excellent communication skills and a Bachelor's degree are required. A competitive salary and benefit package commensurate with experience is offered. Please email a cover letter expressing your research/technical interests and resume to:
Michael A. Mancini, Ph.D. Assistant Professor Director, Integrated Microscopy Department of Molecular and Cellular Biology Baylor College of Medicine Houston, TX 77030 mancini-at-bcm.tmc.edu http://microscopy.bcm.tmc.edu 713 798 8952
Hi Y'All, I'm interested to know what exactly is left behind after Ag (or C) paint (used as an electrical contact on a sample stub) has dried. I have used this method of attaching samples to holders in ultra high vacuum systems and after the paint is thoroughly dried there is no outgassing in uhv to 10-10 torr. However, I wondered if anyone has any idea what the remaining constituents or composition of the paint would be after drying.
Thanks in advance,
Steve
Steve Buckingham Dir. of Process Development Excellatron Solid State LLC 1640 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
Michelle, I have good results staining respiratory mucosal tissue embedded in Araldite 502 with Alcian blue (8GS?) with or without PAS and in conjugation with Weigert's hematoxlyn. The plastic is first etched 1 to 3 minutes with sodium ethoxide, one:one, with toluene. After that I followed standard histological protocols. However, the staining times need to be increased and/or heated to compensate for the reduced number of available staining sites in semithins. The sections were spectacular I don't mind saying.
Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
Hello fellow microscopists-
Does anyone have any experience with staining thick (semi-thin) sections with Luxol Fast Blue?
I believe it's the nature of the Spiroplasma genus membrane structure rather than the poor preservation from your side. The usual glut/OsO4 fixation should be just fine, the problem is in visualization - lack of contrast. There is a good site on http://www.oardc.ohio-state.edu/spiroplasma/what.htm explaining evolutionary degeneration of the cell walls of the whole class Mollicutes. You may want to try some other contrast enhancing methods in addition to traditional heavy metal post-staining. Feel free to contact me off-line.
Alice Dohnalkova Environmental Microbiology Battelle, Pacific Northwest National Lab Richland, WA 99352 (509)372-0692
-----Original Message----- } From: Haixin Xu To: Mel Dickson Cc: microscopy-at-sparc5.microscopy.com Sent: 11/16/2001 9:23 AM
Hi Listers,
We are trying to microscopy a microorganism, spiroplasma using TEM. Every time we had problem to see clearly the membrane around the organism. We tried to increase the time of Osmium (4%) fixation, 3-5 hours at RT after glutaldehyde fixation. But the membrane preservation is still terrible. I would very much appreciate any of suggestions for that.
Steve Buckingham wrote: ========================================================== } I'm interested to know what exactly is left behind after Ag (or C) } paint (used as an electrical contact on a sample stub) has dried. I have used } this method of attaching samples to holders in ultra high vacuum systems and } after the paint is thoroughly dried there is no outgassing in uhv to 10-10 } torr. However, I wondered if anyone has any idea what the remaining } constituents or composition of the paint would be after drying. ========================================================== Our firm has had hands-on invovlement in the formulation of both silver and carbon "paints" for SEM (including UHV) applications going back to the early 1970's. I think I can comment on this topic.
The silver and carbon systems have to be discussed separately.
In the case of the silver system, of the leading brands used in microscopy, there is some considerable difference in the size and shape of the silver colloid, the suspending organic carrier and the composition and loading of polymer(s) that are present to impart to the paint different characteristics . We have learned over the years that some polymer is important in order to give the final paint layer the kind of adhesive properites users desire. More can sometimes be "too much" and some polymers tend to be more effective as an adhesive at low levels than other polymers. The shape of the colloid (e.g. flat platelet vs. round sphere) is important as well, since some sizes/shapes of the silver colloid lead to a higher likelihood of a skin forming during drying, which then acts as a transport barrier so that there is a wet region underneath. When such a skin-over-wet sample is inserted into the vacuum system, we all know the consequences.
Finally, we have found that not all silver paints found in the microscopy market have the same high purity, although we would be the first to recognize that for most SEM users, the presence of such impurities (such as contaminant SiO2 particles, otherwise known as glass frit) at low levels would not make any difference. There is a difference in the % silver loadings, something many just don't appreciate, and sometimes what appears to be a "better price" deal in fact is a product that is "cheaper" because of lower silver solids.......full disclosure on the percent silver solids at least for the SPI Supplies products are given on our website. Further evidence of the differences between products are the differences in flash points as disclosed in MSDS information.
So to now answer the question, with regard to silver, after the liquid carrier evaporates completely, with no wetness under a skin, there should be only the silver colloid, polymer at a low level, and otheriwise unintended impurities. Most people report that when the dried SPI Supplies silver paint is inserted into a UHV sustem, they find no indication of a diminuition of the quality of the vacuum.
For the carbon paint, the system is a bit easier to describe, the colloid is present much as is the silver colloid, a bit of a polymer to give the carbon paint greater adhesion properties, and a low level of impurities. Again, the carbon paints are not "all the same". There are differences in the carbon colloid size, difference in the polymer, and differences in the impurity elements that might be found. When the carbon paint dries, there is also a tendency for a skin to form, and since there are differences in the formulations, there are differences in the tendency for skin formation. But when fully dry, and there is no organic carrier underneath a skin, there should be only the carbon colloid, the polymer, and unintended impurities (which vary from product to product). We have had similar reports with regard to the SPI Carbon Paint product, that when inserted into a UHV system , there is no indication of a diminuiution of the quality of the vacuum.
Disclaimer: For nearly thirty years, SPI Supplies has formulated silver and carbon paints for SEM/EDS and other vacuum applications, and we have been able to do extensive testing on our own in-house equipment of various formulations. Now despite our obvious eagerness to sell more, I would urge consideration of the dry adhesives, mainly the double sided conductive carbon sheets and the analogous double sided silver sheet products. They too seem to be equally vacuum compatible and one eliminates the uncertainly of any kind of "wet" region underneath a skin on the surface. One also eliminates the possibility of inhalation of organic vapors. Such double sided (dry) adhesives are available from SPI Supplies as well as from the main supply sources for consumables for SEM and TEM. But like with the silver and carbon paint products, they don't all "come from the same place" and your experiences with one might not necessarily be the same as with the others. You might want to try more than one kind and then draw your own conclusions.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Erinasilk-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, November 17, 2001 at 15:31:20 ---------------------------------------------------------------------------
Email: Erinasilk-at-aol.com Name: Erin Silk
Organization: Summit Middle Schoo
Education: 6-8th Grade Middle School
Location: Boulder,Colorado,USA
Question: What power microscope do I need to see bacteria grow on samples of meat. I have two microscopes one that is up to 43x and another at up to 900x. I want to see and measure if and how much bacteria is present in my slide samples.
I am looking for a strange device, but I suppose already someone see something like hat.
A friend of mine is an optician and got a slit lamp.
He is now picking up pictures to be stored in a computer with a TV camera.
Obviously, after the frame grabber, he get an ugly quality of the image.
The slit lamp is equipped with a camera attachment and he got a camera too, but the procedure of taking a picture with a film, develop it, scan and store it is too long.
I remember of a converter device that fit in any 35 mm camera with a CCD sensor able to convert a normal film camera in a digital one.
BIMS will soon be profiled by some major analysts and newsletters along with the release of significant news regarding explosive sales for the Company. There will be huge volume and a strong increase in price for several days. The same groups that featured TIWI will begin coverage on BIMS. TIWI exploded from $ .44 to $2.41 in 4 days!! We know for certain that the same groups are going to feature BIMS and even better returns are expected.
We are very proud that we can share this information with you so that you can make a profit out of it. It is highly advisable to take a position in BIMS as soon as possible, today before the market closes or tomorrow.
The stock could easily reach $5.00 in less than a month on the strength of their upcoming contract announcements and Strong Analyst Buy Recomendations.
At 11:09 PM 11/17/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Name: Erin Silk } } Organization: Summit Middle School } } Education: 6-8th Grade Middle School } } Location: Boulder,Colorado,USA } } Question: What power microscope do I need to see bacteria grow on } samples of meat. I have two microscopes one that is up to 43x and } another at up to 900x. I want to see and measure if and how much } bacteria is present in my slide samples. } Erin -
The "900x" may do the job for you. But since anything above 400x SHOULD require what a microscopist calls an "oil immersion objective", I fear that your lenses will not give you a clear image. The best way available to you for actually seeing the bacteria will be to use a sterile swab to take a sample to smear on a slide; you'll probably need to stain them to make them visible. Getting accurate quantitative data probably won't be possible with the methods available to you. Look in catalogs like Flinn Scientific, Carolina Biological, or Edmund Scientific for kits with stains and instructions. If you can't see them, consider trying to grow the bacteria on culture medium in petrie dishes. You can find supplies in the same catalogs.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
This depends on whose paint you use. Ted Pella, SPI, EMS, etc. all use different solvents (such as methylethylketone, acetone, and others), and they use some sort of polymer (acrylic?) as a binder. The best thing is to get the MSD sheet for the particular brand you use, and call the folks from whom you bought the paint.
Phi
} Hi Y'All, } I'm interested to know what exactly is left behind after Ag (or C) } paint (used as an electrical contact on a sample stub) has dried. I have } used this method of attaching samples to holders in ultra high vacuum } systems and after the paint is thoroughly dried there is no outgassing in } uhv to 10-10 torr. However, I wondered if anyone has any idea what the } remaining constituents or composition of the paint would be after drying. } } Thanks in advance, } } Steve } } Steve Buckingham } Dir. of Process Development } Excellatron Solid State LLC } 1640 Roswell Street, Suite J } Smyrna, GA 30080 } phone 770 438 2201 } fax 617 812 5920
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Unless I'm dreadfully mistaken a microtome does not produce extreme heat (like an engine) and would not be used at -100C. The oil is required for lubrication and should cause little drag due to high viscosity. Sounds like sewing machine oil to me. If it was my microtome, I'd use some Molyslip (molybdenum disulfide), which lubricates better than oil, does not lose its properties, sticks better then grease and does not increase drag like grease. Until better reasons are provided, it appears somebody is selling snake-oil. Many years ago a top engineer from Ford Motors compared fancy oil additives with quality engine oil thus: They are mouse milk and very precious, but don't do anything that Cows-milk won't do.
On second thoughts Jonathan: I'll provide twice the volume and include freight cost. Maybe we can corner the world market in "microtome oils". Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, November 17, 2001 7:52 AM, Jonathan Wilson [SMTP:wilson_jm-at-cimar.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I have been loaned a microtome (Leitz 1512) by a colleague which } according to the instruction manual requires regular oiling. The microtome } did not come with any oil and the price I have been quoted by a local } supplier for microtome oil was about 75$US (including VAT) for 50ml (SAKURA } 1447). I found the price a bit steep and am left wondering if it is } absolutely necessary to buy "microtome" oil or if I can use an off the shelf } oil (e.g 3 in 1 or something like that). I do not want to compromise the } performance of the microtome as it is not mine and would appreciate any } advice from those with experience. } Thanks for any help. } Sincerely, } Jonathan } } Jonathan Wilson (PhD) } CIIMAR } Rua do Campo Alegre 823 } 4150-180 Porto, Portugal } tel: 351 22 608 0470 / 71 } fax: 351 22 606 0423 } e-mail: wilson_jm-at-cimar.org } web: www.cimar.org
Hi, I am trying to build fluorescence confocal microscope using a laser,a dichroic mirror, an objective(numerical aperture=0.7;working distance = 6 mm; focal length = 2 mm; focus depth 0.6 micron)which focuses into a cryostat,the emmited light is focused into the monochromator with a mirror in, and then to the CCD.
However, I did not achieve wide illumination, I've checked the spot size with the CCD but it had hardly changed even though i added a short focus lens in the exciting beam path and a diffuser. I also tried to put a longer focusing lens in the exciting beam path so that the focal point will be inside the objective but the image didn't widened.
Does anyone has an idea how to solve the problem and what lens or optics i should use in the path of the exciting laser so that i would get wide illumination. and a wide image as well.
hello everyone, I very rarely send out a blanket message - please bear with me, but you might like to consider this virus warning. best wishes, Jeremy
--- "Sanderson, Yvonne (REPP)" {Yvonne.Sanderson-at-repp.co.uk} wrote: } From: "Sanderson, Yvonne (REPP)" } {Yvonne.Sanderson-at-repp.co.uk} } To: "'jb_sanderson-at-yahoo.com'" } {jb_sanderson-at-yahoo.com} } Subject: FW: New Virus - Important } Date: Mon, 19 Nov 2001 14:19:21 -0000
} Importance: High } } Subject: New Virus - Important } Someone is sending out a very cute screensaver of the Budweiser Frogs. } } } } } } } } } } If you download it, you will lose everything! Your hard } drive will crash } } } } } } } } } } and someone from the Internet will } get your screen name and } } } } } } password! } } } } } } } DO } } } } } } } } } } NOT DOWNLOAD IT UNDER ANY } CIRCUMSTANCES! } } } } } } } } } } It just went into circulation } yesterday. Please distribute } } this } } } } } } } } } } message.This is a new, very } malicious virus and not many } } people } } } } } } know } } } } } } } } about } } } } } } } } } } it. This information was announced } yesterday morning from } } } } } } Microsoft. } } } } } } } } } } Please share it with everyone that } might access the } Internet. } } } } } } } } *************************************************************************************** } The information in this internet eMail is } confidential and is intended } solely for the addressee(s). Access, copying, } dissemination or re-use } of information in it by anyone else is unauthorised. } Any views or opinions } presented are solely those of the author and do not } necessarily represent } those of Reed Educational & Professional Publishing } or any of its affiliates. } If you are not the intended recipient please contact } } Reed Educational & Professional Publishing, Oxford, } +44 1865 311366. } *************************************************************************************** }
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You may try using low molecular weight tannic acid after osmium fixation.
Reference: Nicolae Simionescu and Maia Simionescu 1976. Galloylglycoses of low molecular weight as mordant in electron microscopy. The Journal of Cell Biology. Vol 70, 608-621
Annfook Yang
} } } Haixin Xu {xu-at-gl.umbc.edu} 11/16/01 12:23PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Listers,
We are trying to microscopy a microorganism, spiroplasma using TEM. Every time we had problem to see clearly the membrane around the organism. We tried to increase the time of Osmium (4%) fixation, 3-5 hours at RT after glutaldehyde fixation. But the membrane preservation is still terrible. I would very much appreciate any of suggestions for that.
Microscopy Society of America Undergraduate Research Scholarship Program
With this year's call for applications the MSA Undergraduate Research Scholarship Program begins its 14th year providing funding for undergraduate research. To date over 71 projects covering a wide range of topics in the physical and biological sciences have received support through this program. Over the years nearly all the scholarship recipients have maintained a strong interest in imaging sciences and have gone on to graduate school, professional school, teaching, or industry positions. The program, which is funded by MSA and by matching funds from MSA Sustaining Members, is able to support approximately 30% to 40% of applicants. This past year the MSA Executive Council approved an increased in the amount provided by MSA. This, plus matching funds, will permit funding of additional scholarships for 2002. The maximal award for the Undergraduate Scholarships is $3000 and helps to provide student stipends, supply costs, and limited travel expenses associated with the research. Additional support in the form of instrument use time, equipment purchases, etc. is generally provided by the student's supervisor and/or through the sponsoring institution. Abstracts reporting the research results, are prepared by scholarship awardees, and published in "Microscopy and Microanalysis." The program actively seeks external sources of matching funds in order to maintain the favorable levels of support both in terms of the number of projects supported and the level of support for each. We are extremely grateful for the matching support provided by MSA sustaining members and individuals. Their support over the years has enabled the program to increase both the number of awards and the maximum amount of each award.
The MSA Undergraduate Research Scholarship Program is currently solicting applications from students interested in conducting a research project which involves the use of any microscopy technique. Students should be sponsored by a member of MSA. The maximal award is $3000. The application deadline is Dec 31, 2001. Applications can be obtained from the MSA Business Office, businessoffice-at-msa.microscopy.com or call toll free at (800) 538-3672. If you have any questions or require additional information regarding the program please contact either:
Dr. Ralph Albrecht, University of Wisconsin 1675 Observatory Drive, Madison, WI 53706 (608) 263-3952 or 263-4162; (608) 262-5157 FAX; albrecht-at-ahabs.wisc.edu
Dr. Richard Ornberg, Monsanto Company Analytical Sciences Center, 800 N. Lindbergh Blvd., St. Louis, MO 63167 (314) 694-1184; (314) 694-6727 FAX; rlornb-at-ccmail.monsanto.com
This Year's MSA Undergraduate Research Scholarship Awardees and Alternates were:
Awardees: JORGE GARCIA "Frictional and Mechanical Interactions between Nanotubes and Self-Assembled Monolayers: An Atomic Force Microscope Study" University of Texas-El Paso/Univ. Wisconsin, Madison Advisor: Robert Carpick
JAYAN RAMMOHAN "Imaging Aggregation Proteins InVitro Using Atomic Force Microscopy" Case Western Reserve University Advisor: Steven Eppell
PETER KRSKO "Electron Beam Lithography for Micro-Patterning of Bioactive Surfaces" Stevens Institute of Technology Advisor: Matthew Libera
DANIEL J. MAZEAU "Three-Dimensional Structure of the Human TFIIH-IIE Complex" University of California-Berkeley Advisor: Eva Nogales
DANIEL L. PECHKIS "The Impact of Processing on the Interface between Thin Film Dielectrics and their Silicon Substrates" Southern Connecticut State University Advisor: Christine Broadbridge
EUGENE KUNG "HR-TEM Studies of Polymer-Carbon Nanotube Composites" Princeton University Advisor: Nan Yao
Alternates RENEE LOPEZ SMITH "Comparisons Between Pre-Released and Swimming Sperm Cells of the Fern Lygodium: A Correlated SEM, TEM, and Light Microscope Study" Southern Illinois University Advisor: Karen Renzaglia
ERIN GRUND "Quantitation of Glomerular Cell Number in Diabetic Patients and Normal Controls" University of Minnesota Advisor: John Basgen
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Thanks to all who replied to my posting last week about camera-to-microscope interfacing, and for the suggestions, many of which suggested contacting my local Nikon rep.
I would be perfectly happy to use Nikon parts, after all, I'm trying to arrange a marriage between a Nikon camera and a Nikon microscope, so it shouldn't be THAT difficult.
Unfortunately, in my country there is no direct Nikon presence, only local representatives, but one company handles cameras, another handles microscopes, so the question of their interfacing kind of falls into a small but deep void.
If any expert from Nikon is reading this list, I would be grateful if they would reply to me directly, so that we could engage in a constructive dialogue.
thanx
rtch
} } Hi } } We have a Nikon Labophot-POL microscope. } } In its trinocular head there sits a CF PL 5X Projection lens, above } that is a Microflex HFX-IIA Photomicrographic Attachment, and onto } that is a "35mm camera adapter A", then what is called in the } HFX-IIA manual a "Motorized dark box FX-35WA". The latter looks more } like a camera body to me. } } We want to be able to mount our Nikon D 1 body onto the microscope, } with or without the HFX-IIA, whatever it takes. } } Is there anyone out there who can tell, from the above, what I need } to do to acheive this? } } I would have thought that all we need to do would be to mount the D } 1 body directly onto the trinocular vertical tube, but then I know } almost nothing about light microscopy. }
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I just read Gib Ahistrand's short in "Microscopy Today" about cleaning rolls in a laser printer. When I have problems with rubber rolls, either ink jet printers or laser printers, I rub the rubber with a lintless cloth moistened with isopropanol. It removes the "shine" that Gib is talking about. It definitely fixes the problem on inkjet printers where the paper will not feed from the stack. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Maybe try fast freezing plus freeze substitution? Either high pressure freezing or slam freezing? This might better capture the membrane in its native state. Then a freeze sub into osmium tetroxide in acetone should give good contrast to the membrane.
David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
I am organizing a Leica confocal user/administrator listserver in order to facilitate and encourage dialogue regarding the problems, idiosyncrasies and use of the Leica SP platform for confocal and multiphoton microscopy. I am hoping that such a forum would be regarded as a logical extension of the existing microscopy listserver for Leica confocal users to voice Leica-specific questions/concerns.
A Leica LCSM/MP Listserver will allow us to archive questions and answers, and may offset some of the perceived lack of documentation regarding the software. As I have witnessed recently on the general confocal listserver, Leica is paying attention to users' sharing their dilemmas, concerns and successes. It would be advantageous for system administrators, end users and Leica representatives to continue to build momentum in this direction.
LeicaSP_LCS_MP_Users is a subscriber access list-server devoted to questions and issues unique and/or specific to the Leica SP Series Spectral Confocal and Multiphoton System.
If you would like to subscribe, please send an e-mail message to:
listserv-at-listserv.uiuc.edu
with the body of the message consisting of:
subscribe LEICASP_LCS_MP_USERS {full name}
I will be delighted to manage/maintain a Leica confocal/mp listserver resource. Let me know if you think if you have any thoughts, suggestions or concerns regarding this user forum at your convenience. Thank you.
Best Regards, Karl G.
_______________________________________________ Karl Garsha Specialist in Light Microscopy Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
I work for a company in Lafayette, Louisiana and we have an interest in obtaining some SEM and TEM images of chitin isolated from crab shell waste. Particularly, I am interested if anyone knows of institutions that provide such a service, be it private or public, the cost of such services and contact information. The chitin samples we have are in a flake form about 5 - 7mm square. None of the samples are ready for imaging, so they would need to be prepared for the respective imaging processes. We would prefer the images to be in a digital format.
Thank you in advance for any information received.
André N. Blanchard, Ph.D. Manager-Technical and Business Development The Venture Group, Inc PO Box 53631 Lafayette, LA 70505-3631 U.S.A.
A Leica listserv already exists and the reasons for the establishment of that list several years ago were similar to those being touched upon here. Traffic on that list has been essentially nil for the past two years. This tells me one of three things: a) that nobody has any Leica-specific problems that they feel can be solved by list interaction, b) that the interaction received through the list was too limited to make the list a useful resource, or c) that the list consists of "power users" who fix their own problems. I suspect the answer is a mix of a and c. Certainly a lot of Leica systems have been installed since the list was first put together, and I am the first to admit that it was never well publicized, partly for reasons that may become apparent below.
If you desire to move forward with establishment of this list (and I am not discouraging you from doing so), then I would ask you to consider a few points regarding how listservs work and who stands to benefit from the list. 1) is the list set up as "reply to all" or "reply to sender"? 2) can anyone subscribe (it would appear so) and post, or can only those people with legitimate interests in Leica systems subscribe? 3) will Leica monitor the list? will they respond to posts to the list? These two items are important. With the earlier (still existing) list, the answers are "yes" and "no". I hope you can achieve a "yes" for both questions, but you must at least achieve a "yes" for the first question, otherwise it can be a needlessly unpleasant experience for the company itself - very much a blindside tackle. If people simply want to talk about how terrible tech support or service can be, then the list is pretty useless (witness current BioRad postings) - everyone knows this is a sore point with virtually every fancy piece of equipment sold from microscopes to FACS machines. We don't need "venting" lists, particularly if the people who should be "vented upon" are not listening (if they were, we wouldn't be venting now would we??).
We are about to have a new SP2-MP-UV system installed here. My prediction is that the learning curve on this system will be no steeper than that on the TCS-NT system - fairly shallow but with a few specific headaches (really just finding out which buttons to push when). What would really be useful is a list for people who still have older instruments (e.g., TCS4D) on which the learning curve is not only very steep but also for which the repository of that knowledge is a handful of people worldwide. The final point I would like to bring up is that the hardware/software configurations of the various Leica machines sold over the years (even just SP/SP2 series) can be very different, so it can be dangerous to generalize.
Rob Palmer Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
Hi all, We have been using our Gatan duo mill for some time now with a copper exhaust line on the rotary pump. It has worked fine, but now we have someone who would like to use the iodine guns and I am wondering if we should use a copper exhaust line with a corrosive gas like iodine. Does anyone know if copper is an appropriate material to use for an exhaust line when milling with iodine? Any additional pointers for milling with iodine? TIA. Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
It is difficult to answer the question Maxine Daws raised about the operation of the gun and comumn valves in her electron microscope without having a diagram of the vacuum system; however, operating procedures for a variety of different vacuum systems (with a variety of different kinds of pumps) are described in some detail in Chapter 9 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html and http://pup.princeton.edu/titles/6484.html, for a description). If you were to read this chapter, I believe that you could figure out what the proper operating sequence might be. Otherwise, Maxine, if you will send me a diagram of your system I will see if I can answer your question.
Good luck & Happy Thanksgiving, W. C. Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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For the heck of it I did a quick & dirty EDS (thin window detector) on some old carbon and Ag paint spots, no idea of who distributed this paint. The carbon had significant peaks at C, O & P. The Ag had significant peaks at Ag, C & S. The only surprise to me was the P peak with the carbon paint. The S on the Ag spot probably came from Ag tarnishing in air. For what its worth..... Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Lab University of Kansas
} } Hi Y'All, } } I'm interested to know what exactly is left behind after Ag (or C) } } paint (used as an electrical contact on a sample stub) has dried. I have } } used this method of attaching samples to holders in ultra high vacuum } } systems and after the paint is thoroughly dried there is no outgassing in } } uhv to 10-10 torr. However, I wondered if anyone has any idea what the } } remaining constituents or composition of the paint would be after drying. } } } } Thanks in advance, } } } } Steve } } } } Steve Buckingham } } Dir. of Process Development } } Excellatron Solid State LLC } } 1640 Roswell Street, Suite J } } Smyrna, GA 30080 } } phone 770 438 2201 } } fax 617 812 5920
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA18046 for dist-Microscopy; Mon, 19 Nov 2001 16:59:09 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA18039 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 19 Nov 2001 16:58:39 -0600 (CST) Received: from postal1.lbl.gov (postal1.lbl.gov [128.3.7.82]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA18032 for {microscopy-at-sparc5.microscopy.com} ; Mon, 19 Nov 2001 16:58:26 -0600 (CST) Received: from SpamWall.lbl.gov (localhost [127.0.0.1]) by postal1.lbl.gov (8.11.2/8.11.2) with ESMTP id fAJMsqD26340 for {microscopy-at-sparc5.microscopy.com} ; Mon, 19 Nov 2001 14:54:52 -0800 (PST) Received: from lbl.gov (maokeefe.wins.lbl.gov [131.243.3.225]) by SpamWall.lbl.gov (8.11.2/8.11.2) with ESMTP id fAJMsq526332; Mon, 19 Nov 2001 14:54:52 -0800 (PST) Message-ID: {3BF98EA8.C7832787-at-lbl.gov}
I believe that this is a hoax. See: http://www.f-secure.com/hoaxes/budfrogs.shtml -Mike
Jeremy Sanderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } hello everyone, } I very rarely send out a blanket message - please bear } with me, but you might like to consider this virus } warning. } best wishes, Jeremy } } --- "Sanderson, Yvonne (REPP)" } {Yvonne.Sanderson-at-repp.co.uk} wrote: } From: } "Sanderson, Yvonne (REPP)" } } {Yvonne.Sanderson-at-repp.co.uk} } } To: "'jb_sanderson-at-yahoo.com'" } } {jb_sanderson-at-yahoo.com} } } Subject: FW: New Virus - Important } } Date: Mon, 19 Nov 2001 14:19:21 -0000 } } } Importance: High } } } } Subject: New Virus - Important } } } Someone is sending out a very cute screensaver of the } Budweiser Frogs. } } } } } } } } } } } } If you download it, you will lose everything! Your } hard } } drive will crash } } } } } } } } } } } and someone from the Internet will } } get your screen name and } } } } } } } password! } } } } } } } } DO } } } } } } } } } } } NOT DOWNLOAD IT UNDER ANY } } CIRCUMSTANCES! } } } } } } } } } } } It just went into circulation } } yesterday. Please distribute } } } this } } } } } } } } } } } message.This is a new, very } } malicious virus and not many } } } people } } } } } } } know } } } } } } } } } about } } } } } } } } } } } it. This information was announced } } yesterday morning from } } } } } } } Microsoft. } } } } } } } } } } } Please share it with everyone that } } might access the } } Internet. } } } } } } } } } } } } } } *************************************************************************************** } } The information in this internet eMail is } } confidential and is intended } } solely for the addressee(s). Access, copying, } } dissemination or re-use } } of information in it by anyone else is unauthorised. } } Any views or opinions } } presented are solely those of the author and do not } } necessarily represent } } those of Reed Educational & Professional Publishing } } or any of its affiliates. } } If you are not the intended recipient please contact } } } } Reed Educational & Professional Publishing, Oxford, } } +44 1865 311366. } } } *************************************************************************************** } } } } __________________________________________________ } Do You Yahoo!? } Everything you'll ever need on one web page from News and Sport to Email and Music Charts } http://uk.my.yahoo.com
You need an activated carbon trap between your mechanical pump and your turbo. When I was at the University of Alabama at Birmingham, students on two occasions left the iodine gun valves open. They destroyed the mechanical pump springs and rendered it useless. I found out that we were not the only ones that this happened to. Gatan fixed a bunch of mechanical pumps under warranty and came up with a fix -a carbon trap with a sight glass on the mechanical pump side. (Our second pump failure was not covered.) Inside the tube where you could see, there was a silver strip that would change to black when the trap got saturated. You regenerated the trap by replacing the charge because otherwise you would be getting iodine into the mechanical pump. I would worry that copper might react similarly to iodine as the silver. The iodine does come out the exhaust of the mechanical pump.
I thought that the fix was rather expensive at the time, but it did work. You could probably get a zeolite trap and replace the zeolite with activated carbon from a pet store that sells aquarium supplies. Ten years ago, Gatan wanted $90 for a charge of activated carbon (about a quart jar). You could get twice that amount at a pet store for less than $10.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
-----Original Message----- } From: Greg Strout [mailto:stro8031-at-msmailhub.oulan.ou.edu] Sent: Monday, November 19, 2001 5:17 PM To: Microscopy-at-sparc5.microscopy.com
Hi all, We have been using our Gatan duo mill for some time now with a copper exhaust line on the rotary pump. It has worked fine, but now we have someone who would like to use the iodine guns and I am wondering if we should use a copper exhaust line with a corrosive gas like iodine. Does anyone know if copper is an appropriate material to use for an exhaust line when milling with iodine? Any additional pointers for milling with iodine? TIA. Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
I think you need to check your supply of paints. I never had anything behind C and O on carbon paint, do not remember S on Ag either.
Vladimir
} } For the heck of it I did a quick & dirty EDS (thin window } detector) on some old } carbon and Ag paint spots, no idea of who distributed this } paint. The carbon had } significant peaks at C, O & P. The Ag had significant peaks } at Ag, C & S. The } only surprise to me was the P peak with the carbon paint. The } S on the Ag spot } probably came from Ag tarnishing in air. } For what its worth..... } Bruce } Bruce Cutler, Ph.D. } Director, Microscopy & Electronic Imaging Lab } University of Kansas } } } } } Hi Y'All, } } } I'm interested to know what exactly is left behind } after Ag (or C) } } } paint (used as an electrical contact on a sample stub) has } dried. I have } } } used this method of attaching samples to holders in ultra } high vacuum } } } systems and after the paint is thoroughly dried there is } no outgassing in } } } uhv to 10-10 torr. However, I wondered if anyone has any } idea what the } } } remaining constituents or composition of the paint would } be after drying. } } } } } } Thanks in advance, } } } } } } Steve } } } } } } Steve Buckingham } } } Dir. of Process Development } } } Excellatron Solid State LLC } } } 1640 Roswell Street, Suite J } } } Smyrna, GA 30080 } } } phone 770 438 2201 } } } fax 617 812 5920 } } } } } }
I suggest that you try Structural Research Inc. in Mobil, Alabama, (205) 649-9740. They come recommended although I have no personal experience with them.
Best regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Michael O'Keefe" To: listserver a {MAOKeefe-at-lbl.g {microscopy-at-sparc5.microscopy.com} ov} cc: Subject: Re: Fwd: FW: New Virus - Important
11/19/01 04:58 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I believe that this is a hoax. See: http://www.f-secure.com/hoaxes/budfrogs.shtml -Mike
Jeremy Sanderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } hello everyone, } I very rarely send out a blanket message - please bear } with me, but you might like to consider this virus } warning. } best wishes, Jeremy } } --- "Sanderson, Yvonne (REPP)" } {Yvonne.Sanderson-at-repp.co.uk} wrote: } From: } "Sanderson, Yvonne (REPP)" } } {Yvonne.Sanderson-at-repp.co.uk} } } To: "'jb_sanderson-at-yahoo.com'" } } {jb_sanderson-at-yahoo.com} } } Subject: FW: New Virus - Important } } Date: Mon, 19 Nov 2001 14:19:21 -0000 } } } Importance: High } } } } Subject: New Virus - Important } } } Someone is sending out a very cute screensaver of the } Budweiser Frogs. } } } } } } } } } } } } If you download it, you will lose everything! Your } hard } } drive will crash } } } } } } } } } } } and someone from the Internet will } } get your screen name and } } } } } } } password! } } } } } } } } DO } } } } } } } } } } } NOT DOWNLOAD IT UNDER ANY } } CIRCUMSTANCES! } } } } } } } } } } } It just went into circulation } } yesterday. Please distribute } } } this } } } } } } } } } } } message.This is a new, very } } malicious virus and not many } } } people } } } } } } } know } } } } } } } } } about } } } } } } } } } } } it. This information was announced } } yesterday morning from } } } } } } } Microsoft. } } } } } } } } } } } Please share it with everyone that } } might access the } } Internet. } } } } } } } } } } } } } } ***************************************************************************************
} } The information in this internet eMail is } } confidential and is intended } } solely for the addressee(s). Access, copying, } } dissemination or re-use } } of information in it by anyone else is unauthorised. } } Any views or opinions } } presented are solely those of the author and do not } } necessarily represent } } those of Reed Educational & Professional Publishing } } or any of its affiliates. } } If you are not the intended recipient please contact } } } } Reed Educational & Professional Publishing, Oxford, } } +44 1865 311366. } } } ***************************************************************************************
} } } } __________________________________________________ } Do You Yahoo!? } Everything you'll ever need on one web page from News and Sport to Email and Music Charts } http://uk.my.yahoo.com
Isn't it true some roughing pumps that are intended for this type of application are teflon coated internally for a barrier from corrosive gases?
Scott: By the way, the springs in that pump could have been replaced...
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282
Walck, Scott D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } You need an activated carbon trap between your mechanical pump and your turbo. When I was at the University of Alabama at Birmingham, students on two occasions left the iodine gun valves open. They destroyed the mechanical pump springs and rendered it useless. I found out that we were not the only ones that this happened to. Gatan fixed a bunch of mechanical pumps under warranty and came up with a fix -a carbon trap with a sight glass on the mechanical pump side. (Our second pump failure was not covered.) Inside the tube where you could see, there was a silver strip that would change to black when the trap got saturated. You regenerated the trap by replacing the charge because otherwise you would be getting iodine into the mechanical pump. I would worry that copper might react similarly to iodine as the silver. The iodine does come out the exhaust of the mechanical pump. } } I thought that the fix was rather expensive at the time, but it did work. You could probably get a zeolite trap and replace the zeolite with activated carbon from a pet store that sells aquarium supplies. Ten years ago, Gatan wanted $90 for a charge of activated carbon (about a quart jar). You could get twice that amount at a pet store for less than $10. } } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } P. O. Box 11472 (letters) } Guys Run Rd. (packages) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8515 (fax) } } "The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." } } } } -----Original Message----- } } } From: Greg Strout [mailto:stro8031-at-msmailhub.oulan.ou.edu] } } } Sent: Monday, November 19, 2001 5:17 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Exhaust line for ion milling } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } We have been using our Gatan duo mill for some time now with a copper } exhaust line on the rotary pump. It has worked fine, but now we have } someone who would like to use the iodine guns and I am wondering if we } should use a copper exhaust line with a corrosive gas like iodine. Does } anyone know if copper is an appropriate material to use for an exhaust } line when milling with iodine? Any additional pointers for milling with } iodine? TIA. } Greg } } -- } ================================================================== } Greg Strout } Electron Microscopist, University of Oklahoma } WWW Virtual Library for Microscopy: } http://www.ou.edu/research/electron/www-vl/ } e-mail: gstrout-at-ou.edu } Opinions expressed herein are mine and not necessarily those of } the University of Oklahoma } ================================================================== } } } } }
I was analyzing a situation where some electroless (Cu bath that plates w/o electricity) copper plate was peeling away from a copper substrate.
I have to clues. Oxidation and small blisters.
When I do a tape test I get a continuous sheet of electroless Cu on the back of the tape and also a fresh surface of the Cu substrate. The first clue is that with the naked eye you can see oxidation on both surfaces. To me this means there must have been trapped Oxidation during the plating process. Using SEM/EDS at low and medium kV, I did not observe any contamination on either surface, just very thin oxidation.
The next clue was that when I took a photomicrograph with the polarity of the Everhart and Thornley detector inverted I see a two types of topographies. I see the expected micro-etch of the Cu substrate, but also smooth bumps at 5-10 microns, as if the electroless plated some droplets. I think these droplets are really blisters that occurred at very beginning of the plating process. I feel that the blister cause the poor adhesion. I am not sure what caused the blisters, maybe internal stress in the plated deposit together with oxidized or very thin organic contamination of the substrate. Or possibly gas evolution. The substrate has a very even etch topography across the surface, no bumps. I would gladly e-mail these images to anyone interested in seeing this unexpected use of a replicated surface. I would also appreciate any comments.
We are looking into buying a Nikon coolpix 995 digital camera to be used to take mostly brightfield images in a Zeiss Axioplan microscope. So I would like to know how good this camera is for photomicrography. I also found out Nikon has a new coolpix 5000 in the market. Does anyone have good experience with this new one for our kind of application ?
Thanks a lot,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
I would like to receive any information about review papers on the basic principles of histochemical techniques. I am quite interested to receive references of papers on the basic characteristics of a histochemical method (reproducibility, specificity etc).
Many thanks.
Fabio Grizzi Scientific Direction Istituto Clinico HUMANITAS Milan, Italy
I would like to get in touch with Tina to ask her a question. I cannot seem to find her email address. Can anyone help?
Better yet, Tina, if you're listening, could you please reply?
Thanks.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
There is no such thing... Recently we have more and more spams and hoaxes, unfortunately. This virus warning (and the others) is a big fat hoax (http://www.symantec.com/avcenter/venc/data/buddylst.zip.html), only just for clogging the net, generating chain-mails... If you receive any "Important-Virus warning" in the future, please check the Symantec page for hoaxes first (http://www.symantec.com/avcenter/hoax.html). Don't forward these messages automatically, especially not for lists or discussion forums... Regards,
Imre
Imre Kovacs M.D., Ph.D. Research Fellow Genetics and Aging Unit Harvard Medical School Massachusetts General Hospital East Building 114, 16th Street Charlestown, MA, 02129-4404 Phone: 617-724-3253 Fax: 617-724-1823
On Tue, 20 Nov 2001 gary.m.brown-at-exxonmobil.com-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Michael, } } I suggest that you try Structural Research Inc. in Mobil, Alabama, (205) } 649-9740. They come recommended although I have no personal experience with } them. } } Best regards, } } Gary M. Brown } ExxonMobil Chemical Company } Baytown Polymers Center } 5200 Bayway Drive } Baytown, Texas 77520-2101 } phone: (281) 834-2387 } fax: (281) 834-2395 } e-mail: Gary.M.Brown-at-ExxonMobil.com
"Histological and Histochemical Methods" by John Kiernan
Most (all?) editions of "Cell and Tissue Biology" by Leon Weiss (editor) have a nice chapter on histochemistry by Helen Padykula.
Grizzi Fabio ICH wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } } I would like to receive any information about review papers on the basic } principles of histochemical techniques. I am quite interested to receive } references of papers on the basic characteristics of a histochemical method } (reproducibility, specificity etc). } } Many thanks. } } Fabio Grizzi } Scientific Direction } Istituto Clinico HUMANITAS } Milan, Italy
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reimar_gaertner-at-wsib.on.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, November 19, 2001 at 14:48:41 ---------------------------------------------------------------------------
Question: My daughter and I have come across a most curious pond-water creature that we can't identify in any of our books. It is large - about 400 um in diameter and round. It has movable internal organs that move food around - especially two pincer-like claws that appear to be macerating captured food. It therefore must be an animal - no cilia are apparent. It has a unique method of feeding: it unfolds a clear satalite-dish-like catchers mit with which it directs passing prey into it's mouth area. It can quickly retract the antena array when touched or jolted. What is this marvelous thing?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (elir-at-chem.ch.huji.ac.il) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 18, 2001 at 13:55:49 ---------------------------------------------------------------------------
Email: elir-at-chem.ch.huji.ac.il Name: Eli rothenberg
Organization: Hebrew University
Education: Graduate College
Location: Jerusalem, Israel
Question: I would like to know what is the correct setup for a wide illumination in a confocal microscopy arrangement, that is, what optics should i use in the path of the laser, if i want to get the maximum illuminated area through my objective, the setting is as follows: laser-collimated beam- dichroic mirror-objective-illuminated surface. (p.s I thought of using a short focusing lens that will focus half way through the objective so maybe that will give me a wide illumination on the surface, and/or maybe a diffuser to go along with it?)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (malone1527-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 18, 2001 at 19:17:02 ---------------------------------------------------------------------------
Email: malone1527-at-aol.com Name: Gene Cimino
Organization: None-Microscopy is a hobby for me. I am 58 years old
Education: Undergraduate College
Location: Hydes,MD,USA
Question: Is Balsam still considered a good mounting medium? If so, can it be kept fluid enough to apply by adding Xylol to it? I am curious about this as I want to make some mounts of insect structures. I know about Permount, but I wanted to try the older Balsam method.By the way,I got my BS in Biology in 1967 from Towson University.
It sounds to me very much like a rotifer: these are very common, and can survive considerable desiccation, and so are often found in roof gutters (where I remember finding them). I would guess that the "satellite dish" is the ciliated "wheel" from which the phylum gets its name - I think that the cilia are either moving too fast or simply too small to be seen at whatever magnification you are using. This generates a current, and I remember as a boy watching green spherical algae being sucked in and macerated.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
----- Original Message ----- } From: {reimar_gaertner-at-wsib.on.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, November 21, 2001 3:34 AM
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I am trying to find out what has been recently published in the field of ink-jet materials (dyes, pigments) using transmission electron microscopy. If you know of any reviews or research articles on this specific subject I would appreciate your feedback!
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Pedro Miguel Rodrigues de Almeida, Dr. Sc. Agfa-Gevaert N.V. RDM/PA-Mo - Electron Microscopy Laboratory Septestraat 27 BE-2640 Mortsel
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Dear expert microscopist, I'm a very beginner with SEM imaging and particularly with EDS Microanalysis. I'm trying to work with my own standard (Oxford ME-....) and the standard-loading job is not quite simple for me. I've got I few questions tha you might find stupid but I would appreciate any help!!! 1) The SEM is a LEO 4400 2) I work with geological sample 3) The EDS software package is ISIS 300 4) What are the best conditions for loasing the standard? i.e. 15 or 20 Kv EHT? 600 ma I-Probe or more? 50 or 100 sec. Preset Time? I have mostly pure metals and compounds plus Natural Albite, Ortoclase and Wollastonite. 5) Once I've performed the calibration using Cobalt, must I fix the counts per second to the same value I obtained for the Cobalt for any further standard acquisition? 6) Before preparing the element profiles, what's the best way to strip peaks away from the spectrum: basic strip or advanced strip? This is particularly important for silicates where Si, Al and Na peaks are close (i.e. whithin the -600/+600 ev energy windows). 7) How can I perform deconvolution of peaks (Si and Al) before saving the profiles? Is it possible to "erase" unwanted peaks from the standard spectrum without altering the background? Thank You for any answer or suggestion. I will appreciate. Melina Meli
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id XAA06086 for dist-Microscopy; Wed, 21 Nov 2001 23:35:31 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id XAA06083 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 21 Nov 2001 23:35:00 -0600 (CST) Received: from bloodwood.adelaide.edu.au (bloodwood.adelaide.edu.au [129.127.43.1]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id XAA06076 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 21 Nov 2001 23:34:48 -0600 (CST) Received: from adelaide.edu.au ([129.127.101.59]) by bloodwood.adelaide.edu.au (Netscape Messaging Server 4.15) with ESMTP id GN6SMM00.90Z for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 22 Nov 2001 16:00:22 +1030 Message-ID: {3BFC8D51.E5B293F-at-adelaide.edu.au}
Hi all,
I have been asked to remove a specimen from a sem stub turn it over and photograph the other side. The specimen is an organically preserved plant fossil and very fragile, it is attached to the sem stub using double sided black tape called STR tape made by Shinto Chemitron Co. Ltd. I have tried soaking a piece of the tape in acetone and also in ethanol to see if this would dissolve the tape. Neither did anything and the tape is still sticky. Any suggestions would be greatly appreciated
Jo Shrapnel Environmental Biology Adelaide University
Dear Listers, As my wife and I drove and hiked around the Northeast one last time to see the fall colors before driving across from New York to California, I thought of the differences among what could be seen while walking, driving, and flying. It occurred to me that these modes of transportation differed in rates by roughly factors of ten: a brisk walk is about 2 m/s; a leisurely drive is about 20 m/s; an airliner travels at about 200 m/s. As I thought further, I realized that a weather satellite at a height of about 1000 km would have an orbital speed of about 2 km/s, and scanning through a TEM grid looking for suitably crystallized outer mitochondrial membranes would happen at a speed of about 2 um/s. (I can examine the ~10 cm screen at 40 kx in about 1 s.). Also, a trip across the solar system at 0.01 c would take about a day. Other possibilities for filling in this series could be the scan speed of AFM on one end and the rate that the Hubble telescope can record a field for the deep field survey at the other. (Someone knowledgeable in astronomy could calculate the apparent speed by figuring how much distance is represented by the angular width of the Hubble field of view at a depth of 10^10 light-years and dividing by the exposure time.) Can we put together a complete list of powers-of-ten speeds, along with how these can be realized and what typically can be seen? I'll start.
?? AFM individual atoms 2 um/s TEM organelles 2 m/s walk individual rocks, trees, streams, etc. 20 m/s drive mountains, lakes, forests, etc. 200 m/s fly mountain ranges, storms, deserts, etc 2 km/s orbit weather patterns, continents 200 Mm/s ion drive? solar system ?? Hubble scan quasar field I hope you will have as much fun with this as I have. Yours, Bill Tivol
I'm not familiar with that particular product, but adhesives that remain sticky (rather than dry out) usually respond very well to naptha. Lighter fluid for zippo type cigarette lighters contains naptha and is an inexpensive and readily available source.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
-----Original Message----- } From: Joanne Colmer [SMTP:joanne.colmer-at-adelaide.edu.au] Sent: Wednesday, November 21, 2001 9:30 PM To: Microscopy-at-sparc5.microscopy.com
Hi all,
I have been asked to remove a specimen from a sem stub turn it over and photograph the other side. The specimen is an organically preserved plant fossil and very fragile, it is attached to the sem stub using double sided black tape called STR tape made by Shinto Chemitron Co. Ltd. I have tried soaking a piece of the tape in acetone and also in ethanol to see if this would dissolve the tape. Neither did anything and the tape is still sticky. Any suggestions would be greatly appreciated
Jo Shrapnel Environmental Biology Adelaide University
Dear all Most photographers are only too aware that flash intensity diminishes according to the square of the distance to the subject, making it difficult to use flash for distance photogrpahy with a lens or telescope. Some years ago I considered the possibility of illuminating a subject viewed with a telescope or telephoto lens by using the same optics to focus an electronic flash on the subject. A problem was how short would the flash duration have to be for the all illuminating light to have left the optics before image-forming light started to come back. I concluded that for an object distance of 50m, i.e. a total light-path of 100m, would be about a third of a microsecond, and gave up at that point, having no way of generating flashes that short.
An orders of magnitude table (in 3-orders of magnitude steps) for distance travelled by a light flash looks like this: 1 second 300,000 km 1 millisecond 300 km (this is typical electronic flash duration) 1 microsecond 300 m 1 nanosecond 0.3 m 1 picosecond 0.3 mm 1 femtosecond 0.3 µm (30nm)
so a light "beam" of 1 femtosecond duration, such as can be produced by lasers, is in fact a light disc!
Not a lot of people know that :-) Best wishes Chris
Date sent: Thu, 22 Nov 2001 03:10:18 -0500
Joanne
Chloroform appears to soften our sticky tape and its adhesive within a few minutes - if you have any spare STR tape you could try it first. It should be easy to separate the sample then. However I don't know whether chloroform will damage your sample, you will probably need to rinse the surface of the sample with fresh solvent and remember to use the chloroform in the fume hood.
Malcolm Haswell e.m. unit, School of Sciences, University of Sunderland, UK (+44)0191 515 2872
Joanne Colmer wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi all, } } I have been asked to remove a specimen from a sem stub turn it over and } photograph the other side. } The specimen is an organically preserved plant fossil and very fragile, } it is attached to the sem stub using double sided black tape called STR } tape made by Shinto Chemitron Co. Ltd. } I have tried soaking a piece of the tape in acetone and also in ethanol } to see if this would dissolve the tape. } Neither did anything and the tape is still sticky. } Any suggestions would be greatly appreciated } } Jo Shrapnel } Environmental Biology } Adelaide University
Dear Colleagues, A colleague of mine, working on a thesis in pathology is looking for review papers on the application of confocal microscopy in cancer research, diagnosis and prognosis. Can anyone cite useful recent references for him, or possesses or has authored a favored reference? many thanks, Jeremy Sanderson
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First posting to the list. I originally posted this on Sci.techniques.microscopy but was advised that better response may be obtained here. I'm a post doc reseacher at the Uni of B'Ham, UK.
I was wondering whether anybody had any experince of using colloidal gold in ESEM. I wish to visualise fungal extracellular matrices released onto glass or plastic surfaces in "wet" mode. If any body has any advice on this, or has used colloidal gold in ESEM and can pass on a few tips it would be most appreciated.
Regards
Steve Thomas
********************************* Dr Steve Thomas School Of Biosciences The University of Birmingham Edgbaston Birmingham B15 2TT 0121 414 5573 Email: S.Thomas-at-bham.ac.uk
Jo Shrapnel wrote: ======================================================= I have been asked to remove a specimen from a sem stub turn it over and photograph the other side. The specimen is an organically preserved plant fossil and very fragile, it is attached to the sem stub using double sided black tape called STR tape made by Shinto Chemitron Co. Ltd. I have tried soaking a piece of the tape in acetone and also in ethanol to see if this would dissolve the tape. Neither did anything and the tape is still sticky. Any suggestions would be greatly appreciated ======================================================== The adhesive system will not really "dissolve" in any common solvents. It can be swollen and loosened, such as in chloroform, which can aid in its removal, but it will not actually be dissolved away in the traditional sense . For a flat surface often times such solvent treatment is enough but for your kind of sample, I would trust it would not be enough.
But in a sense you are fortuante in that your sample of interest is not organic but is in fact inorganic in nature. You should be able to put it into a small glass petri dish, without cover, contaminating adhesive side "up", and then using an oxygen plasma, such as would be generated in the SPI Supplies® Plasma Prep™ II plasma etcher, the organics should be etched away and the substrate (relatively) untouched. I say "relatively" because eventually, in the limit, the oxygen will cause changes in the substrate, but the etch rate for the organic adhesive is many times faster than for the inorganic substrate. So you want to be careful to etch only to the extend you need to etch and to try not etching beyond that point.
Special note and disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher so we would naturally have a vested interest in expanding its use. However, the same physics found in the SPI Plasma Prep II unit also exist in several other table-top systems being manufactured such as by Denton Vacuum and Polaron (Now Quorum Technologies, formerly, VG Microtech). More about the Plasma Prep II plasma etcher can be found at URL http://www.2spi.com/catalog/instruments/etchers1.html
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We are trying to locate a journal in the medical or cellular fields with an online version that accepts videoclips. Thank you in advance for your help.
as there was a little interest in the Leonid meteors on this list server last week I thought would report my observations from Ayers Rock, Australia.
Central Australia was expected to be reasonably placed to observe the peak activity early on Monday morning, Nov 19th. The sky was clear and dark from sunset to sunrise and relatively warm (about 16°C). From the sand dunes I spent a very pleasant 5 or 6 hours under the stars and counted approximately 300 meteors in the first 90 minutes. After that the rate was too great to keep count. I estimate seeing approximately 2000 to 3000 meteors, with many others that I missed because I was looking the wrong way at the time. I can well imagine whiplash injuries being commonplace amongst those of us fortunate enough to be in the right place at the right time with the right weather conditions.
By comparison, the next morning I saw only 8 meteors in 90 minutes, and only 2 of these were Leonids.
Nestor, I managed to capture a few meteors in photos (one 15 minute exposure has over 20 meteors visible on the print). I'll bring some of the better ones to Adelaide. Cheers,
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} From: Maike Andersson [m.andersson-at-t-online.de] Sent: Mittwoch, 14. November 2001 21:27 To: Microscopy
I am trying to characterize the water pathway in plant tissue by the water soluble, apoplastic tracer sulforhodamin. In order to prevent secondary movement of the tracer, I want to freeze substitute under anhydrous conditions prior to embeddment in Spurr. What substitution media do you recommend? Does polar acetone lead to loss or secondary movement of the tracer? Is apolar diethylether a better choice and how can I overcome the very slow rewarming protocols of it? Thanks in advance
Maike Andersson Institut for Applied Botany Hamburg / Germany
Listers; The University of Calgary has a terrific job opening in the Microscopy and Imaging Facility.
Please check it out. Calgary is one of the most prime cities to live in. We are 70 miles from Banff and the best mountain activities in the world. Our province has no sales tax!
Because of government regulations we must first look at qualified Canadian citizens or Landed Immigrants and then folks from other nations. We of course are an equal opportunity employer.
Jon McGovern Manager, Microscopy and Imaging Facility University of Calgary Calgary, Alberta, Canada.
Position Available
The Microscopy and Imaging Facility of the University of Calgary requires an experienced EM technologist. The Microscopy and Imaging facility houses three transmission and three scanning electron microscopes. The flagship of these being a newly acquired FEI/Philips Tecnai F20 equipped with EDX, STEM, EELS, and Cryo imaging. This instrument is the only one of its kind in Canada. The successful candidate should have a very strong background in TEM, preferably with cryo microtomy and cryo observation. Experience in operational instruction to a wide group of users is a necessity. Overseas training on the Tecnai F20 will be provided. It is expected that the successful candidate will assist with the SEM side of the facility as required. Although there are no strict educational requirements, candidates should possess the appropriate qualifications and training for the position. Experience in operating EM’s, routine maitenance, management of the day-to-day operation of the instruments, the ability to instruct new users, and work in a multi-user environment would be an asset. This position will appeal to persons who thrive on challenges and are willing to take on demanding tasks. Applicants are invited to forward their resumes, three letters of reference and if possible examples of their expertise to:
Dr. John Reynolds (reynolds-at-ucalgary.ca) Head, Department of Cell Biology and Anatomy Faculty of Medicine University of Calgary 3330 Hospital Dr NW Calgary, Alberta Canada T2N 1N4
I've never looked at colloidal gold in an ESEM before, although I've imaged it in a FESEM, but my immediate reaction is that you would have two things working against you. One is that to image a wet or uncoated sample, you would have to introduce sufficient gas into the chamber to seriously degrade your resolution. The second is that you need to use the backscatter detector to image the gold particles, as you know, which again is significantly lower resolution than the secondary imaging mode. With these two resolution-losers combined, I seriously doubt that you could image gold particles down into the nanometer range.
But, that said, I always tell people that only way to really know for sure is to try. But, is there some compelling reason why the sample must be viewed in "wet" mode, or for that mattter, in an SEM at all? Is there a reason not label and embed your samples for TEM?
Good luck, Randy Tindall EM Core University of Missouri Columbia, MO
-----Original Message----- } From: Steve Thomas To: Microscopy-at-sparc5.microscopy.com Sent: 11/22/01 10:28 AM
Hello,
First posting to the list. I originally posted this on Sci.techniques.microscopy but was advised that better response may be obtained here. I'm a post doc reseacher at the Uni of B'Ham, UK.
I was wondering whether anybody had any experince of using colloidal gold in ESEM. I wish to visualise fungal extracellular matrices released onto glass or plastic surfaces in "wet" mode. If any body has any advice on this, or has used colloidal gold in ESEM and can pass on a few tips it would be most appreciated.
Regards
Steve Thomas
********************************* Dr Steve Thomas School Of Biosciences The University of Birmingham Edgbaston Birmingham B15 2TT 0121 414 5573 Email: S.Thomas-at-bham.ac.uk
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bob-at-rockisland.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, November 24, 2001 at 15:06:36 ---------------------------------------------------------------------------
Email: bob-at-rockisland.com Name: Bob Carter
Organization: independant researcher
Education: Graduate College
Location: Lopez Island WA 98261 USA
Question: Hi. I recently purchased a Vickers M171237 (Patholux???). It is a compound "research " type similar to the Leitz Orthomat. It has a quick change 5 objective nose, digital readout, substage and epi lighting, plus a box under the binocular with a beamsplitter, pol, and other optics I am unfamiliar with. I am restoring my Vickers for daily usage. I am looking for technical information, and user or repair guides. I completely disassembled, cleaned, reassembled, and adjusted an AO MicroStar, but this microscope is far more complicated. If you know of any resources, it would be greatly apreceated. Thanks, Bob
Bob Carter 2000 Bayshore Road Lopez Island, WA USA 98261
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Is there any direct way to quantitatively determine and measure the actual spot size of a SEM beam?
Given various KV, WD, final aperture sizes, condenser lens settings, etc., is there a way to truly find out what the real spot size is on the specimen?
In TEM we measure the actual contamination spot "burned" into the carbon film by the beam. I use a grid with calibration spheres for reference. With the appropriate organic film, a similar technique could work with SEM in spot mode?? I'm not sure how much accuracy you're looking for, but it might give you an idea.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Sunday, November 25, 2001 8:59 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser } lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g.
Gary, One technique I've seen used is to mount an aperture over a hole (on carbon is best), or better yet, a thinned foil for a sharper edge, and use a line-profile mode to scan across after focussing at a very high mag. Absorbed current can be used instead of secondary for a signal, although the latter works well, especially on a thinned foil. The curve you get will resemble a rise-time curve and can be treated in a similar manner. Mark the 10% and 90% rise points on the curve, then measure the horizontal distance between those two points in units appropriate for the mag. This is generally considered to be the beam diameter given the Gaussian distribution of electrons across the beam.
Ken Converse owner Quality Images third party SEM service Delta, PA
Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g. } } } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (twiinie-at-aolcom) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 25, 2001 at 22:36:35 ---------------------------------------------------------------------------
Email: twiinie-at-aolcom Name: Brett Chamberlin
Education: 6-8th Grade Middle School
Location: Gresham, Oregon USA
Question: We recently purchased a fairly old Bushnell Microscope. It is all metal construction with a service sticker. The service sticker reads a last serviced date of 1986.
Where might I find a manual and/or some instruction on the features of this microscope. There are some attachments that I do not understand the function of.
It says "Magna" on the front of it.
Thank you very much for any help that you can offer.
Any calculation would require factors specific for any one make of instrument. Anyway, doing it experimentally is more fun. Microanalysts need to know the actual location of the spot on the specimen and for that observe the beam spot on a daily basis. For WDS an SEM needs among other accessories a light microscope. With that and with the beam on a fluorescent mineral the beam spot is readily visible.
Usually in SEM the spot is busy scanning, so spot mode is required and for that the SEM scan coils are switched off. (Also, turn down fluorescent screen/ monitors, to avoid burn-marks. With a graticule in the eyepiece and micron- step stage movements the beam can be measured, but . . . For small spots, suitable for SEM the microscope on a microprobe will not have enough power.
The alternative is to burn spots onto a smooth specimen in a location which could be found under a high power light microscope. A grided coverslip with a light carbon film may be suitable. By parking the beam spot near line intersections the burn mark could later be found and measured. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Monday, November 26, 2001 11:59 AM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g.
This method has promise....it is not too difficult to do! Sounds sort of like a Faraday cup affair. I could try this with 35u, 70u and 100u apertures mounted on a Carbon stub.
In general terms, what IS a typical spot size? Is it Angstroms, microns, or what? I never really thought about the true spot size--only relative, based on CL setting. Higher setting, smaller spot size. If a spot is used with the SEM in spot mode, the issue is to find out how big the spot actually is. This affects spillover of data when doing x-ray probing of very thin layers in microchips.
thanks to all, gary g.
At 05:53 AM 11/26/2001, you wrote: } Gary, } One technique I've seen used is to mount an aperture over a hole (on } carbon is best), or better yet, a thinned foil for a sharper edge, and use } a line-profile mode to scan across after focussing at a very high } mag. Absorbed current can be used instead of secondary for a signal, } although the latter works well, especially on a thinned foil. The curve } you get will resemble a rise-time curve and can be treated in a similar } manner. Mark the 10% and 90% rise points on the curve, then measure the } horizontal distance between those two points in units appropriate for the } mag. This is generally considered to be the beam diameter given the } Gaussian distribution of electrons across the beam. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Gary Gaugler wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi: I am looking to replace my old Epson flat bed scanner and want a high quality scanner. This instrument is used primarily for scanning TEM negatives, 35 mm slides, some OCR and needs to be function in a multi-user environment. I have been considering the UMAX Powerlook 3000 and the Heidelberg Linoscan 2650 due to their reported 3.6 dynamic range and 42 bit pixel depth. Does anyone have experience with these machines? Is there something better?
Thanks!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
Dear Gary, The classic method is to take the highest magnification photo that will show you the resolution limit, defined as the closest two points can be and still be determined as two points instead of one. I just take a hi-mag photo of a highly detailed surface and measure the smallest detail I can see and divide by the magnification. This will be the resolution at the conditions currently set. I was always told that the resolution of the SEM was equal to the diameter of the electron beam. For a sample I use an evaporated gold film on a graphite substate, to reduce the contribution of back-scattered electrons. At 05:59 PM 11/25/01 -0800, you wrote: } } Hi all: } } Is there any direct way to quantitatively determine } and measure the actual spot size of a SEM beam? } } Given various KV, WD, final aperture sizes, condenser lens } settings, etc., is there a way to truly find out what } the real spot size is on the specimen? } } Any ideas? } } gary g. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters.
Thanks, Mike
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
I have _usually_ assumed for purposes of x-ray analysis that the SEM spot size is zero. I know it is not, but considering the interaction volume inside the specimen, the beam size is almost negligible. The various Monte Carlo programs tell me that I must consider an interaction volume on the order of a micron or more at 15 kV depending on my substrate. I am pretty sure that my beam is not half a micron in diameter, so how much will it affect the interaction volume?
I suppose the experiment to try would be to use an x-ray signal along with the secondary or absorbed current signal to check the effective spot size. And I suppose I should check the backscattered signal too while I am at it.
"Fortunately", much of my work has not involved the small structures where we are striving to keep the beam within the very thin layers. But those problems are coming along with increasing frequency. One of these days I will probably need to delve into the deconvolutions needed to determine the compositions of phases smaller than the excitation volume. I recall seeing an example of principle component analysis at MSA (2000?) where spectrum imaging allowed the discernment of a thin phase at an interface that was ordinarily undiscernible. Of course I do not have the program or equipment that those folks at Oak Ridge and NORAN had, so it will be a while before I could repeat the experiment. I wonder if anyone has developed a good, public-domain version of the routine.
Warren
At 06:20 AM 11/26/01 -0800, Gary wrote:
} Ken: } } This method has promise....it is not too difficult to do! } Sounds sort of like a Faraday cup affair. I could try } this with 35u, 70u and 100u apertures mounted on } a Carbon stub. } } In general terms, what IS a typical spot size? Is it } Angstroms, microns, or what? I never really thought } about the true spot size--only relative, based on CL setting. } Higher setting, smaller spot size. If a spot is used with } the SEM in spot mode, the issue is to find out how big } the spot actually is. This affects spillover of data when } doing x-ray probing of very thin layers in microchips. } } thanks to all, } gary g. } } } At 05:53 AM 11/26/2001, you wrote: } } Gary, } } One technique I've seen used is to mount an aperture over a hole (on } } carbon is best), or better yet, a thinned foil for a sharper edge, and } } use a line-profile mode to scan across after focussing at a very high } } mag. Absorbed current can be used instead of secondary for a signal, } } although the latter works well, especially on a thinned foil. The curve } } you get will resemble a rise-time curve and can be treated in a similar } } manner. Mark the 10% and 90% rise points on the curve, then measure the } } horizontal distance between those two points in units appropriate for the } } mag. This is generally considered to be the beam diameter given the } } Gaussian distribution of electrons across the beam. } } } } Ken Converse } } owner } } Quality Images } } third party SEM service } } Delta, PA } } } } Gary Gaugler wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for a hydrophobic heavy metal stain to stain the hydrophobic core of micelles suspended in water. I need to view the micelles with TEM. Non-water soluable hydrophobic drug molecules partition rapidly into the core; and I suspect a hydrophobic heavy metal such as a stable organo metallic would also, such as lead ethylhexanoate. Any suggestions or experience you can share with me?
We would like to dimple silver for TEM sample prep. When we used our standard methods, it dimpled exceedingly slowly (perhaps because it is soft).
Does anyone have experience in dimpling silver successfully? -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Is this a silver film on a substrate of some sort or is it a lump of Ag? If it is a piece of silver why don't you electroplish it? Far fewer artifacts.
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com NOTE CHANGES: 1-845-892-2225 Office; -6899 FAX; 1-800-352-4732, pin 8604 Pager; 1-914-453-2917 Personal Cell Phone
Alwyn Eades {jae5-at-lehigh.edu} on 11/26/2001 01:15:31 PM
To: EMNET {microscopy-at-sparc5.microscopy.com} cc:
We would like to dimple silver for TEM sample prep. When we used our standard methods, it dimpled exceedingly slowly (perhaps because it is soft).
Does anyone have experience in dimpling silver successfully? -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This is a good point; the interaction volume overwhelms the actual "spot size" under most circumstances. But since Gary asked for some actual numbers: In TEM (W filament) I generally find the spot ~200nm or less by measuring the contamination spot. As a matter of interest, I believe for NVLAP asbestos analysis a spot {250nm is required. That is, the *minimum* obtainable spot must be {250nm. Since, as Ken mentioned, the distribution is presumed Gaussian there is a bit of subjectivity as to where you pick the "edge" and how long you burn....not to mention time-dependent drift.
I have a table of "theoretical" spot sizes for my ESEM-FEG: they range from {1nm to about 40nm depending on kV and spot size selected.
Matt
On Monday, November 26, 2001 11:20 AM, Warren E Straszheim [SMTP:wesaia-at-iastate.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } I have _usually_ assumed for purposes of x-ray analysis } that the SEM spot } size is zero. I know it is not, but considering the } interaction volume } inside the specimen, the beam size is almost negligible. } The various Monte } Carlo programs tell me that I must consider an interaction } volume on the } order of a micron or more at 15 kV depending on my } substrate. I am pretty } sure that my beam is not half a micron in diameter, so how } much will it } affect the interaction volume? } } I suppose the experiment to try would be to use an x-ray } signal along with } the secondary or absorbed current signal to check the } effective spot size. } And I suppose I should check the backscattered signal too } while I am at it. } } "Fortunately", much of my work has not involved the small } structures where } we are striving to keep the beam within the very thin } layers. But those } problems are coming along with increasing frequency. One } of these days I } will probably need to delve into the deconvolutions needed } to determine the } compositions of phases smaller than the excitation volume. } I recall seeing } an example of principle component analysis at MSA (2000?) } where spectrum } imaging allowed the discernment of a thin phase at an } interface that was } ordinarily undiscernible. Of course I do not have the } program or equipment } that those folks at Oak Ridge and NORAN had, so it will be } a while before I } could repeat the experiment. I wonder if anyone has } developed a good, } public-domain version of the routine. } } Warren } } At 06:20 AM 11/26/01 -0800, Gary wrote: } } } Ken: } } } } This method has promise....it is not too difficult to do! } } } } Sounds sort of like a Faraday cup affair. I could try } } this with 35u, 70u and 100u apertures mounted on } } a Carbon stub. } } } } In general terms, what IS a typical spot size? Is it } } Angstroms, microns, or what? I never really thought } } about the true spot size--only relative, based on CL } } setting. } } Higher setting, smaller spot size. If a spot is used } } with } } the SEM in spot mode, the issue is to find out how big } } the spot actually is. This affects spillover of data } } when } } doing x-ray probing of very thin layers in microchips. } } } } thanks to all, } } gary g. } } } } } } At 05:53 AM 11/26/2001, you wrote: } } } Gary, } } } One technique I've seen used is to mount an aperture } } } over a hole (on } } } carbon is best), or better yet, a thinned foil for a } } } sharper edge, and } } } use a line-profile mode to scan across after focussing } } } at a very high } } } mag. Absorbed current can be used instead of secondary } } } for a signal, } } } although the latter works well, especially on a thinned } } } foil. The curve } } } you get will resemble a rise-time curve and can be } } } treated in a similar } } } manner. Mark the 10% and 90% rise points on the curve, } } } then measure the } } } horizontal distance between those two points in units } } } appropriate for the } } } mag. This is generally considered to be the beam } } } diameter given the } } } Gaussian distribution of electrons across the beam. } } } } } } Ken Converse } } } owner } } } Quality Images } } } third party SEM service } } } Delta, PA } } } } } } Gary Gaugler wrote: } } } } } } } ------------------------------------------------------- } } } } ----------------- } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } } } Society of America } } } } To Subscribe/Unsubscribe -- Send Email to } } } } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } } } } } } } } http://www.msa.microscopy.com/MicroscopyListserver/FA Q.htm } } } } l } } } } ------------------------------------------------------- } } } } ----------------. } } } } } } } } Hi all: } } } } Is there any direct way to quantitatively determine } } } } and measure the actual spot size of a SEM beam? } } } } Given various KV, WD, final aperture sizes, condenser } } } } lens } } } } settings, etc., is there a way to truly find out what } } } } the real spot size is on the specimen? } } } } Any ideas? } } } } gary g. } }
I am trying to find information on remote control and remote viewing on the SEM. I am currently trying to use Microsoft Net Meeting. Does anybody have a better way to do these things?
Is there a reliable method of detecting Anthrax spores within a very short time, through the use of Electron Microscopy? Do you know anyone working with this method of detection of Anthrax spores? The system I am looking into involves using sticky pads to capture spores in work areas to include heating and cooling vents. The material collected is then taken to an area where a microscope is used to determine the presence or no-presence of the spores. Within a few hours, 2-4 hours, there is a determination made if anthrax spores are in fact present.
which also presents some other solutions. This is a pdf copy of the slides I presented during a tutorial at the M&M2000 meeting. That tutorial was video taped so if you are really keen you can order a copy of the tape from the MSA Education Committee.
} Email: twiinie-at-aolcom } Name: Brett Chamberlin } } Education: 6-8th Grade Middle School } } Location: Gresham, Oregon USA } } Question: We recently purchased a fairly old Bushnell Microscope. It } is all metal construction with a service sticker. The service sticker } reads a last serviced date of 1986. } } Where might I find a manual and/or some instruction on the features } of this microscope. There are some attachments that I do not } understand the function of. } } It says "Magna" on the front of it. } } Thank you very much for any help that you can offer. } ---------------------------------------------------------------------------
Brett -
If you can't find a manual, the general advice in this book will be very helpful:
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy; it's unequalled as a basic reference for beginners. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with good illustrations) of commonly encountered organisms. Adult. RECOMMENDED
The description above was taken from the MICRO bibliography; you'll find other helpful books there. URL below.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
In a message dated 11/26/2001 10:23:39 AM US Mountain Standard Time, herro001-at-umn.edu writes:
{ { I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters. } }
Mike,
Is the autofluorescence yellowish? Rather than try and scrub a mouse pup, I would first try filtering out the offending autofluor color with the scope, assuming it probably isn't at exactly the same wavelength as the GFP signal.
It depends on your scope and what kind of filters you are using, but I would contact either the manufacturer of the scope or the fluorescence rig and inquire about narrow bandpass filters for GFP (both excitation and emission). Some filter sets have longpass filters in them that pass the required signals plus everything on either side for quite a distance out.
If you can get narrow bandpass filters, this should help a lot. If you already have narrow bandpass filter in your scope...never mind the above advice! I wonder how baby shampoo works on newborn mice...??
FYI, we have had good success using the advice of the folks at Chroma Technology Corp., especially Paul Millman. You can contact Paul at Choma by calling (800) 824-7662 in the U.S. They're experts at everything that's fluorescence-related.
Good Luck!
Bob Chiovetti GTI Microsystems Tucson, AZ Leica Exclusive Regional Dealers (AZ, NM, West TX)
Would appreciate some help with preparation of gram -ve bacteria for SEM.
I plan to do a standard gold coating but was wondering if anyone has some practical experience on fixation before proceeding with the rest of the protocol.
Many thanks in advance.
Lachlan
________________________ Lachlan McDonald EnGeneIc Pty. Ltd. 105 Delhi Road Riverside Corporate Park North Ryde NSW 2113 AUSTRALIA
} Hi, } } We are trying to salvage our 8 year old Cambridge Instruments Quantimet 570 } image analysis workstation. However there is no sign of any manuals. Does } anybody have a manual that they would be willing to photocopy for us, } donate, orcan you put me in contact with someone who might have one?? } } Any help appreciated, } } Jim } } } Jim Buckman } Department of Petroleum Engineering } Heriot-Watt University } Riccarton } Edinburgh } EH14 4AS } Scotland } } jim.buckman-at-pet.hw.ac.uk
I don't have experience with either of the two scanners you mentioned. For our customers, we usually recommend Microtek. We have had good success with the (previously discussed here) ArtixScan 2500 which is 36-bit with a 3.4 dynamic range. Resolution is 2500 dpi (optical) and the list price is $4,499. (Usually available for less.) (10,000 element CCD)
Microtek also offers (but I have yet to use) the ArtixScan 1100 which is closer to the ones you mentioned. Optical resolution is 2000 x 1000, 42-bit, and a higher reported dynamic range of 3.9. This one has a list price of $1,499. (8,000 element CCD)
Both of these use dual plate technology including Microtek's patented Emulsion Direct Imaging Technology for incredibly sharp transparency scanning. With E.D.I.T., there is no glass between the scanning lens, film, or light source, eliminating distortion and Newton rings that occur with traditional transparency adapters.
I hope this helps. If I can be of any additional assistance, please let me know.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045
-----Original Message----- } From: Ken Bart [mailto:kbart-at-hamilton.edu] Sent: Monday, November 26, 2001 10:09 AM To: microscopy-at-sparc5.microscopy.com
Hi: I am looking to replace my old Epson flat bed scanner and want a high quality scanner. This instrument is used primarily for scanning TEM negatives, 35 mm slides, some OCR and needs to be function in a multi-user environment. I have been considering the UMAX Powerlook 3000 and the Heidelberg Linoscan 2650 due to their reported 3.6 dynamic range and 42 bit pixel depth. Does anyone have experience with these machines? Is there something better?
Thanks!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
Mike, Why don't you try something like magic mending tape and do a few strips on the areas of interest. The tape is quit good at cleaning surfaces and it should be gentle enough and leave no residue. Good Luck, Russ Gillmeister Xerox ~~~~~~~~~~~~~
-----Original Message----- } From: Michael Herron [mailto:herro001-at-umn.edu] Sent: Monday, November 26, 2001 12:15 PM To: microscopy-at-sparc5.microscopy.com
All,
I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters.
Thanks, Mike
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
The answer is complex. Probably, the bottom line for practicality is "no", not easily or rapidly. It could be done with the right equipment and the right reagents.
Bacillis anthrasis spores morphologically resemble spores of other Bacillus species (e.g., B. cereus). Some of these other bacilli are ubiquitous (e.g., on food). Thus, it would be impossible to differentiate, based solely on appearance, what species the spores were.
One possibility would be to react an antibody against B. anthrasis with them and then tag that first antibody with a secondary antibody that had a marker recognizable in the electron microscope. This could be done either by scanning electron microscopy (SEM) of the surface of spores, or by transmission electron microscopy (TEM) of ultrathin sections or of negatively stained bacteria--although negative staining would not be ideal because of the large size of the organism. (Negative staining is usually used for small things like viruses).
This immunolabeling technique would not be particularly fast (a matter of hours, rather than the days it would take to grow and speciate the bacteria, but not minutes). It would also be rather cumbersome in requiring immunological reagents and operators familiar with immunolabeling and thin sectioning. It would also require that the bacteria be handled as a biohazard and be killed before examination to prevent contamination.
Another possibility for identification, or for at least preliminary indication that malice was intended, would be to examine the preparation with Xray microanalysis. Most times, weaponized B. anthrasis spores are mixed with a chemical that causes them not to form large clumps. (The smaller particle size allows them to get further into the small airways of the lung.) This powder could be detected with microprobe analysis. However, one could question whether a smart crook might use other kinds of powder with different components to disguise the dispersant. Also, one would need to kill the spores while working on them to prevent infection.
I hope this helps. While I¹m in the business of using the electron to diagnose infectious diseases (we examine almost a thousand samples a year for viruses), and welcome any new use, I want to acknowledge the limitations.
Sara Miller
(P. S. The genus and species should be either italicized or underlined; my primitive email program won¹t do this.)
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
On Mon, 26 Nov 2001, Norton, Buddy wrote:
} Date: Mon, 26 Nov 2001 19:35:31 -0600 } From: Norton, Buddy {hnorton-at-richmond.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: detecting Anthrax spores using EM? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is there a reliable method of detecting Anthrax spores within a very short } time, through the use of Electron } Microscopy? Do you know anyone working with this method of detection of } Anthrax spores? The system I am looking into } involves using sticky pads to capture spores in work areas to include } heating and cooling vents. The material collected is then } taken to an area where a microscope is used to determine the presence or } no-presence of the spores. Within a few hours, 2-4 } hours, there is a determination made if anthrax spores are in fact present. } } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } "Buddy" } } Howard B. Norton } Captain } Deputy Chief of Police - Operations } } Office: 804-289-8723 } Pager: 804-346-6263 } Fax: 804-289-8720 } } hnorton-at-richmond.edu } } 31 UR Drive } Special Programs Building } University of Richmond Police Department } University of Richmond, Virginia 23173 } } For Sale - 1985 Prowler Camper Trailer 32' } } Those who row the boat seldom have time to rock it. } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } }
Our lab. had a failure of a thermal imaging system. The system is used for thermal mapping integrated circuits. We are interested in using a contract service to do measurements for us while our hardware is being repaired. Does anyone know of a contract lab., preferably on the East Coast, that has this capability for hire? I may consider a thermal AFM if available. The system we have now is optical and low resolution compared to an AFM. Resolution is ~ 10 uM.
Regards, Peter Tomic Failure Analysis & Analytical Services Group Anadigics, Inc. Warren, New Jersey
The New England Society for Microscopy (NESM) will be holding it's 35th Annual Fall Symposium at UMASS-Lowell in Lowell, Massachusetts on Friday, Dec. 7, 2001.
The meeting will begin at 12 Noon with Registration in the Mil Conference Center-Wannalancit Building (North Campus).
There will be 3 Sessions: A special student session beginning at 1:05 pm with 4 presentations, followed by a Poster session and coffee break.
Session II (Biological) begins at 2:20 pm and has as it's speaker, Ken Moore (University of Iowa), the National MSA Speaker. His talk will be: "Application of Microscopy Techniques to the Study of Genetic Therapy Research".
Another Poster Session and Afternoon Coffee Break will follow at 3:20 pm.
Session III (Materials Science) will begin at 3:40 pm and have 2 speakers: Elen Humphreys from M.I.T. and Koichi Nishikida from Thermo Spectra-Tech.
The Annual Business Meeting will convene at 5:00 pm and will include the election of NESM officers for 2002.
A Dinner and after-dinner speaker will follow at 6 pm.
Advance registration is due by Friday, November 30th. Registration after November 30th will NOT include dinner. The registration for NESM members is $10.00 and $25.00 for non-members. Dinner is $15.00 extra.
For more detailed information, please contact Mary McCann, NESM Treasurer at mccanns-at-tiac.net or NESM's website: The New England Society for Microscopy (NESM) will be holding it's 35th Annual Fall Symposium at UMASS-Lowell in Lowell, Massachusetts on Friday, Dec. 7, 2001.
The meeting will begin at 12 Noon with Registration in the Mil Conference Center-Wannalancit Building (North Campus).
There will be 3 Sessions: A special student session beginning at 1:05 pm with 4 presentations, followed by a Poster session and coffee break.
Session II (Biological) begins at 2:20 pm and has as it's speaker, Ken Moore (University of Iowa), the National MSA Speaker. His talk will be: "Application of Microscopy Techniques to the Study of Genetic Therapy Research".
Another Poster Session and Afternoon Coffee Break will follow at 3:20 pm.
Session III (Materials Science) will begin at 3:40 pm and have 2 speakers: Elen Humphreys from M.I.T. and Koichi Nishikida from Thermo Spectra-Tech.
The Annual Business Meeting will convene at 5:00 pm and will include the election of NESM officers for 2002.
A Dinner and after-dinner speaker will follow at 6 pm.
Advance registration is due by Friday, November 30th. Registration after November 30th will NOT include dinner. The registration for NESM members is $10.00 and $25.00 for non-members. Dinner is $15.00 extra.
For more detailed information, please contact Mary McCann, NESM Treasurer at mccanns-at-tiac.net or NESM's website: http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
There is a SEM/EDS software "gunshot inspection" for particle inspection automatically. I use this system for particle analysis on hard drive disk, a 95 mm diameter disk with 0.5 um resolution in whole disk scaning. The particle size range can be set so that the software ignores the larger one or the smaller one (dirt), and locates the particles and does EDS analysis automatically. I do not think you need to do EDS, but the image capture and classification functions may be good enough for you.
Contact Oxford Instrument (I have this system) or Noran Instrument (they make the similar system) for more information. Your sample should be no problem (particles on sticky pad).
Hope this helps,
Best regards,
Zhiyu Wang Sr. Engineer Maxtor Corp. Milpitas, CA (408)432-4421
-----Original Message----- } From: zhiyu wang [mailto:zhiyuw-at-home.com] Sent: Monday, November 26, 2001 2:39 PM To: zhiyu_wang-at-maxtor.com
The problem with this method is that most Bacillus species spores will look alike by SEM . . . and Bacillus species are very common contaminants. Most Bacillus species are not pathogenic. Some of the species spores, such as Bacillus stearothemophilus are used on purpose, like in Spore Strips, which are put into autoclaves to make sure the proper kill time and temp are reached. Unless there are some very distinctive characteristics only found on Bacillus anthracis spores, SEM or any other strictly visual method will not work to prove the presence of anthrax-causing spores.
Dr.Tina S. Schwach, President Microscopy Consulting Services Inc. 651-681-0112
----- Original Message ----- } From: "Norton, Buddy" {hnorton-at-richmond.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, November 26, 2001 7:35 PM
As discussed by Sara Miller and Tina Schwach, the genus Bacillus can not be distinguished on the basis of morphology alone. One needs specific tests: (1) immunological tests such as fluorescent antibody staining and light microscopy (LM) or even antibody-labeled latex spheres that would attach to the spores (LM and EM), (2) molecular amplification techniques where unique sequences are amplified and identified. Either of these tests take hours rather than days. The gold standard is still growing the organisms from the spores since even a single spore is detectable by this method. The downside is that culturing can take many days.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
With this instrumentation, one might be able to detect particles the size of bacillus spores, but would not be able to tell the difference between spores of the different bacillus species.
Sara Miller
On Tue, 27 Nov 2001, Wang, Zhiyu wrote:
} Date: Tue, 27 Nov 2001 09:45:59 -0700 } From: Wang, Zhiyu {Zhiyu_Wang-at-Maxtor.com} } To: "'Norton, Buddy'" {hnorton-at-richmond.edu} } Cc: "'microscopy-at-MSA.Microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Subject: RE: detecting Anthrax spores using EM? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, Norton: } } There is a SEM/EDS software "gunshot inspection" for particle inspection } automatically. I use this system for particle analysis on hard drive disk, } a 95 mm diameter disk with 0.5 um resolution in whole disk scaning. The } particle size range can be set so that the software ignores the larger one } or the smaller one (dirt), and locates the particles and does EDS analysis } automatically. I do not think you need to do EDS, but the image capture and } classification functions may be good enough for you. } } Contact Oxford Instrument (I have this system) or Noran Instrument (they } make the similar system) for more information. Your sample should be no } problem (particles on sticky pad). } } Hope this helps, } } Best regards, } } Zhiyu Wang } Sr. Engineer } Maxtor Corp. } Milpitas, CA } (408)432-4421 } } -----Original Message----- } } From: zhiyu wang [mailto:zhiyuw-at-home.com] } Sent: Monday, November 26, 2001 2:39 PM } To: zhiyu_wang-at-maxtor.com } Subject: Fw: detecting Anthrax spores using EM? } } } } ----- Original Message ----- } } From: "Norton, Buddy" {hnorton-at-richmond.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, November 27, 2001 1:35 AM } Subject: detecting Anthrax spores using EM? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Is there a reliable method of detecting Anthrax spores within a very short } } time, through the use of Electron } } Microscopy? Do you know anyone working with this method of detection of } } Anthrax spores? The system I am looking into } } involves using sticky pads to capture spores in work areas to include } } heating and cooling vents. The material collected is then } } taken to an area where a microscope is used to determine the presence or } } no-presence of the spores. Within a few hours, 2-4 } } hours, there is a determination made if anthrax spores are in fact } present. } } } } } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } } } "Buddy" } } } } Howard B. Norton } } Captain } } Deputy Chief of Police - Operations } } } } Office: 804-289-8723 } } Pager: 804-346-6263 } } Fax: 804-289-8720 } } } } hnorton-at-richmond.edu } } } } 31 UR Drive } } Special Programs Building } } University of Richmond Police Department } } University of Richmond, Virginia 23173 } } } } For Sale - 1985 Prowler Camper Trailer 32' } } } } Those who row the boat seldom have time to rock it. } } } } $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Endospores are produced by many bacteria in the genera Bacillus and Clostridium. Many of these bacteria are quite common and most are harmless. EM could doubtless identify a bacterial endospore as such but could not, by itself, identify the species as Bacillus anthracis. Even so, a positive result should elevate your typical "suspicious white poweder" to the status of a "very credible biohazard threat" and elicit a vigorous response from the FBI, CDC etc.
Phase contrast light microscopy would be just as capable for this purpose, and much more convenient than EM. LM and EM would also both provide some characterization of dispersants mixed with the spores (i.e. bentonite, etc.).
Sample preparation and microscopy should be done with appropriate biosafety containment measures. Anthrax requires Biosafety II containment with special attention to elimination of aerosols.
I have designed sampling and inspection procedures for our campus. If you need further details you can contanct me off list.
At 07:35 PM 11/26/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Hi I was wondering if any one has a good electropolish recipe, using a double jet-polisher to thin a niobium sample, with out using HF if possible. The sample is needed for high resolution TEM imaging.
Thanks,
Cindy Kleier EPIC Facility Northwestern University j-kleier-at-northwestern.edu
Mike, In my experiments with skin imaging, the dryer the skin (and flakes) the brighter the autofluorescence. How would the pups respond to a little hand lotion? (no experience with this, do lotions autofluoresce?)
As an aside, has anyone else observed that the yellow stratum corneum autofluorescence (in unfixed skin cross sections) actually increases with time exposed to the light beam? Is this just localized drying, or something else (the sections are mounted in 50% glycerol)? Added fluorescent dyes photobleach as expected. Karen -------------------------------------------------------------------------------- Karen S. Zaruba BioMaterials Technology Center 3M Center Bldg. 270-1S-01 Tel: (651) 737-2971 Fax: (651) 737-5645
Michael Herron {herro001-at-umn.edu} 11/26/01 11:15 AM Please respond to herro001
To: microscopy-at-sparc5.microscopy.com cc: (bcc: Karen S. Zaruba/US-Corporate/3M/US) Subject: Whole body fluorescence microscopy
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All,
I am using a stereo microscope equipped for epiflourescent GFP excitation and emission. I am looking at live GFP transgenic mice and while the technique works quite well, there is a problem with autoflourescence of the skin surface.
These are mouse pups with little or no hair, and the skin has dander or flakes that seems to autofluor. I was wondering if anyone might know of a solution I could quench this external autofluor with? Something mild would be in order since these are delicate little critters.
Thanks, Mike
--
________________________________________________________ / Michael J. Herron, U of MN, Dept. of Pediatrics/BMT / / herro001-at-umn.edu / / 612-626-4321 Mpls MN 55455 / /_______________________________________________________/
I have gotten an old Cooke Vacuum Products CV 301 evaporator to restore but have no manuals. I received an email 'manual' on how to operate the vacuum system valves but not how to connect the transformer connections behind a clear plastic panel to do an evaporation.
There are six wing nuts with connections on a phenolic mounting board behind a plastic panel and two copper bus bars can be connected about seven different ways. I believe you connect the bottom right and bottom middle wing nuts together with one bus bar (the common ground). That connects the two 6 volt center taps together with the two right hand feed throughs in the bell jar. Also the top middle nut probably swings to either of the two bell jar connections in order to do an evaporation. Just guessing.
Volt meter readings could not sort out what the big step down transformer internal wiring was.
Please email me privately if you know how to connect them without me shorting the transformer out. The 2 transformer secondary tap SETS are labeled zero 0, 6 and 12 (volts). I would be glad to call you for your advice.
Any operating manual concerning the proper connection sequence would be nice to get in the mail. The manufacturer says he has none for this old instrument.
Paul Beauregard Senior Research Associate PPG Industries 440 College Park Drive Monroeville, PA 15146
Ron, Right you are! Oops! Actually, the beam could be even wider if the distibution wasn't Gaussian. Guess this requires a more expert source of information.
Ken Converse owner Qualtiy Images third party SEM service Delta, PA
Ronald Anderson wrote:
} Ken, } } You are only seeing the rise part of the beam width distribution curve. } Wouldn't the distance between the points be the beam radius and not the } beam diameter? } } Ron } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } NOTE CHANGES: 1-845-892-2225 Office; -6899 FAX; 1-800-352-4732, pin 8604 } Pager; 1-914-453-2917 Personal Cell Phone } } }
Hi All, In summer it is difficult to get nice Formvar-carbon films for electron microscopy grids. So we prefer to prepare them in autumn or winter, because the humidity is less important in our laboratory. My question is, according your experiences, how many times can we conserve the Formvar-carbon coated grids and at which temperature.
Thank you very much,
Robert Alain Microscopie Electronique Inrs-Institut Armand-Frappier Laval, H7V 1B7 Robert.alain-at-inrs-iaf.uquebec.ca
I have full paper technical data for q570, but it is too long to you, I have this email for q570 "ia-support-at-leica.co.uk", ask people on this adress (Richard Hayden ?), or find Leica www page and call to him with this problem, this q570 is very good image analyser
best regards
Krzysztof Jan Hübner Foundry Reserach Institute, ul. Zakopianska 73 Kraków, Poland
Hi, Niobium is polished with good results in an acid and water free solution (H and O atoms are absorbed by unprotected Nb surfaces) using a solution of 10g Magnesium Perchlorate (Mg[ClO4]2)in 1 liter Methanol (Struers Tenupol: 60 V; 85 mA; temperature -5°C; flow rate 5); ref.: T. Schober and V. Sorajic, Metallography 6 (1973) 183. Good luck, H.K. Schmid
Please, please, please DO NOT take sampling and analysis of suspected bioattack materials into your own hands. Contact the authorities. If you are indeed dealing with an agent you have a "hot zone" and have to be in full protective gear. As someone already mentioned, you have to do the work at the highest BSL the unknown might contain. Small quantities of B.anthracis would be BL2 but what if it turns out to be something worse? You are risking turning your labs into hot zones. Also you are dealing with a crime scene or a potential crime scene. Anything you do could be contaminating evidence and jeopardizing the probability of conviction. I know it is tempting to play detective on your own, but there is an infrastructure in place for dealing with these things. Anthrax is not the only agent "out there" and a course of antibiotics isn't going to do the trick for all of them.
My intention is not to slam Buddy and Mike for their comments, as for all I know they are part of the infrastructure and are working with appropriate authorities in what they do. I just don't want anyone to get it into their heads to go sleuthing into potential incidents with an "it's okay, I'm a scientist" attitude. Even the people here, who design biodetectors and train first responders across the country, call in the authorities if they suspect anything.
Dr. Erica R. Valdes US Army Edgewood Chemical Biological Center EC/B Forensic Analytical Center AMSSB-RRT-CF/Building 3160 Aberdeen Proving Ground, MD 21010
-----Original Message----- } From: Mike Dalbey [mailto:dalbey-at-biology.ucsc.edu] Sent: Tuesday, November 27, 2001 2:27 PM To: Microscopy-at-sparc5.microscopy.com
Endospores are produced by many bacteria in the genera Bacillus and Clostridium. Many of these bacteria are quite common and most are harmless. EM could doubtless identify a bacterial endospore as such but could not, by itself, identify the species as Bacillus anthracis. Even so, a positive result should elevate your typical "suspicious white poweder" to the status of a "very credible biohazard threat" and elicit a vigorous response from the FBI, CDC etc.
Phase contrast light microscopy would be just as capable for this purpose, and much more convenient than EM. LM and EM would also both provide some characterization of dispersants mixed with the spores (i.e. bentonite, etc.).
Sample preparation and microscopy should be done with appropriate biosafety containment measures. Anthrax requires Biosafety II containment with special attention to elimination of aerosols.
I have designed sampling and inspection procedures for our campus. If you need further details you can contanct me off list.
At 07:35 PM 11/26/01 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
} } } } } } Hi All, } } } } } } I have been given a set of blocks of mouse brain (corpus callosum) to } } } cut. The investigator did her own embedding in Embed-812/Araldite } } } and the blocks are actually quite nice at first blush. The } } } semi-thins are gorgeous and I can cut nice light gold-silver } } } sections, but her boss is interested in the interline structure of } } } the myelin sheaths, and wants silver-grey sections. The block won't } } } cut well that thin. I get alternating silver then gold-purple } } } sections and sections either stick to the knife edge or drag over. I } } } have tried all the standard tricks: a newly re-sharpened diamond } } } knife, smallest possible block face, faster cutting speed. I've } } } played with the meniscus. I even put the blocks back in the oven for } } } another day. We will get whatever we can out of these blocks (the } } } result of a many-week treatment), but want to know how she should } } } vary her formula to get harder blocks next time around. She is using } } } a pretty standard recipe (Mollenhauer's Mixture No. 1, 1964). It is: } } } } } } Embed 812 25 ml } } } Araldite 15 ml } } } DDSA 55 ml } } } DMP-30 1.5% } } } } } } } } } I know that there are other recipes out there, but since I'm a } } } Spurr's fan, I'm not familiar enough with them to know which ones } } } will yield what we need. We probably need just a tad more hardness } } } to enable the really thin sections. } } } } } } Thanks in advance, } } } Lee } } } -- } } } Leona Cohen-Gould, M.S. } } } Sr. Staff Associate } } } Director, Electron Microscopy Core Facility } } } Manager, Optical Microscopy Core Facility } } } Joan & Sanford I. Weill Medical College } } } of Cornell University } } } voice (212)746-6146 } } } fax (212)746-8175
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I looked up two in the Philips Number 5 book that have no HF.
Schrober and Sorajic, 1973, Metallagraphy,6,p.183. 0.05 molar magnesium pechlorate per liter of methanol. 50-70V, -5 deg C.
and another Gidley and Davis, 1967 , J Sci. Instrum. 44, p. 297. 90% methanol 10% perchloric 5.5V, -30C BE CAREFUL WITH Perchloric acid solutions!!!
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: J. Cindy Kleier [mailto:j-kleier-at-northwestern.edu] Sent: Tuesday, November 27, 2001 3:51 PM To: microscopy-at-sparc5.microscopy.com
Hi I was wondering if any one has a good electropolish recipe, using a double jet-polisher to thin a niobium sample, with out using HF if possible. The sample is needed for high resolution TEM imaging.
Thanks,
Cindy Kleier EPIC Facility Northwestern University j-kleier-at-northwestern.edu
We are interested in differentiating between arteries and veins in the adult rat pulmonary vasculature. Are there specific markers or antibodies we could use? A variety of plant lectins exists which label endothelial cells but the literature is confusing about their specificities. They may also preferentially stain only certain organs.
We are perfusing the rats, and cutting thick sections (~100 microns) on a vibratome for confocal microscopy. So staining could be carried out either at the time of perfusion or later, with the sections. Is there enough time for a probe to stick during perfusion? A fluorescent marker is required for the confocal technique.
I look forward to any suggestions. Thank you
Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8 Canada
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} Hi All, } In summer it is difficult to get nice Formvar-carbon films for electron } microscopy grids. So we prefer to prepare them in autumn or winter, } because the humidity is less important in our laboratory. } My question is, according your experiences, how many times can we } conserve the Formvar-carbon coated grids and at which temperature. } } Thank you very much, } } Robert Alain } Microscopie Electronique } Inrs-Institut Armand-Frappier } Laval, H7V 1B7 } Robert.alain-at-inrs-iaf.uquebec.ca
I have kept C coated formvar grids for negative staining for a couple of years in an a/c lab. These usually have a medium to "thickish" C coat. I have prepared these in the summer as well, and eastern KS is fairly humid. Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Lab University of Kansas
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Hi Judy,
Just a thought: Could you use a fluorescent marker for endothelial cells such as CD-31 (PeCam) and then use the autofluorescence of the elastin sheath that would be around the arteries to distinguish the ateries from the veins?
Bob Underwood U of Washington Seattle
On Wed, 28 Nov 2001, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow Microscopists: } } We are interested in differentiating between arteries and veins in the adult rat pulmonary vasculature. Are there specific markers or antibodies we could use? A variety of plant lectins exists which label endothelial cells but the literature is confusing about their specificities. They may also preferentially stain only certain organs. } } We are perfusing the rats, and cutting thick sections (~100 microns) on a vibratome for confocal microscopy. So staining could be carried out either at the time of perfusion or later, with the sections. Is there enough time for a probe to stick during perfusion? A fluorescent marker is required for the confocal technique. } } I look forward to any suggestions. } Thank you } } Judy } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital, 8Q } 30 Bond St. } Toronto, ON M5B 1W8 } Canada } } ph: 416-864-6060 x6337 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } }
Hi Listers, I would appreciate any PRIVATE feedback I can get from USERS of the following:
JEOL JEM 1230 Hitachi 7500 or 7600 Philips Technai 12 LEO 910 Quartz Imaging Intranetworking and Image Database Technology [URL:http://www.quartzimaging.com/index.html]
Thanks for the help to all.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
We have a paper in our archive of technical reports titled "A Jet Electropolishing Solution for Silicon Germanium, Tantalum, Niobium, and Tungsten Rhenium" written by Bernie Kestel. This is for our Model 550 Single Vertical Jet Electropolihser, but it may be useful anyway. I will put a copy in the mail to you.
You can view other papers and application notes on our website at www.southbaytech.com.
I hope this helps.
David
"J. Cindy Kleier" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi I was wondering if any one has a good electropolish recipe, using a } double jet-polisher to thin a niobium sample, with out using HF if } possible. The sample is needed for high resolution TEM imaging. } } Thanks, } } Cindy Kleier } EPIC Facility } Northwestern University } j-kleier-at-northwestern.edu
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
One possibility would be to make the Formvar films whenever the humidity permitted it and then coat them later (weeks, months) as needed. For small particulate particles (e.g., viruses, subcellular organelles) adherence is better with freshly carbon-coated grids. Alternativily, you could glow discharge the stored Formvar and carbon coated grids prior to use to improve spreading and adherence.
Sara Miller
On 28 Nov 2001, Bruce Cutler wrote:
} Date: 28 Nov 2001 10:20:38 -0600 } From: Bruce Cutler {bcutler-at-eureka.idl.ukans.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: TEM Conservation of Formvar-carbon coated grid } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Hi All, } } In summer it is difficult to get nice Formvar-carbon films for electron } } microscopy grids. So we prefer to prepare them in autumn or winter, } } because the humidity is less important in our laboratory. } } My question is, according your experiences, how many times can we } } conserve the Formvar-carbon coated grids and at which temperature. } } } } Thank you very much, } } } } Robert Alain } } Microscopie Electronique } } Inrs-Institut Armand-Frappier } } Laval, H7V 1B7 } } Robert.alain-at-inrs-iaf.uquebec.ca } } } I have kept C coated formvar grids for negative staining for a couple of years } in an a/c lab. These usually have a medium to "thickish" C coat. I have prepared } these in the summer as well, and eastern KS is fairly humid. } Bruce } Bruce Cutler, Ph.D. } Director, Microscopy & Electronic Imaging Lab } University of Kansas } } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I agree that sampling and analysis of "suspicous white powders" is not a casual activity and that it must be done with serious consideration of potential risks. Our activity is fully integrated with existing hazmat response procedures followed by the campus police and EH&S. This includes sample collection by EH&S personnel with appropriate personal protection.
Our procedure was created following an incident on campus that the authorities (i.e. FBI) could not effectively respond to because they were overwhelmed by such a large number of similar incidents in the aftermath of the anthrax exposures on the east coast. The FBI agent that did eventually respond had zero knowledge of microbiology and zero personal protection gear. I suspect that waiting for a response by someone prepared to deal with "the highest BSL the unknown might contain" (that would be level IV, I assume) would mean we would be waiting still, and that a major campus building would still be closed. (That material proved to be laundry detergent, by the way.) Based on this experience we concluded that, in fact, there was NOT an infrastructure in place to deal with these situations AT THE LEVEL WE ARE CURRENTLY EXPERIENCING THEM. We hope and expect this will be a very temporary situation.
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Dear Listers, I am preparing a report comparing digital cameras for TEMs. We are looking to upgrade our system intending to image negative stains of heterochromatin samples. At this point we do not have cryo capability. If any of you have experience with the digital cameras which can be upgraded for automated tomography, I would appreciate hearing from you. Rosemary Walsh
The incident we had here at Oklahoma State was handled Veterinary medicine and the State Veterinary Diagnostic lab on campus. The Haz Mat guys showed up in moon suits but vet med didn't. They came in rubber boots, mask, coveralls, goggles, gloves and cap the same as they would for a necropsy of a suspicious animal.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger } } } I agree that sampling and analysis of "suspicous white powders" is not a } casual activity and that it must be done with serious consideration of } potential risks. Our activity is fully integrated with existing hazmat } response procedures followed by the campus police and EH&S. This includes } sample collection by EH&S personnel with appropriate personal protection. } } Our procedure was created following an incident on campus that the } authorities (i.e. FBI) could not effectively respond to because they were } overwhelmed by such a large number of similar incidents in the aftermath of } the anthrax exposures on the east coast. The FBI agent that did eventually } respond had zero knowledge of microbiology and zero personal protection } gear. I suspect that waiting for a response by someone prepared to deal } with "the highest BSL the unknown might contain" (that would be level IV, I } assume) would mean we would be waiting still, and that a major campus } building would still be closed. (That material proved to be laundry } detergent, by the way.) Based on this experience we concluded that, in } fact, there was NOT an infrastructure in place to deal with these } situations AT THE LEVEL WE ARE CURRENTLY EXPERIENCING THEM. We hope and } expect this will be a very temporary situation. } } Mike Dalbey } } Lecturer in Biology } University of California, Santa Cruz } } 831-459-3674 } } dalbey-at-biology.ucsc.edu } } }
How many anthrax cases have we had in the United States in the last 50 years?
} From January 1955 to December 1999, there were 236 reported cases of anthrax, most of them cutaneous, in 30 states and the District of Columbia.
When was the last case of inhalational anthrax in the United States?
The last case of inhalational anthrax in the United States, before 2001, was in 1976 in California. A home craftsman, who worked with yarn, died. Bacillus anthracis was isolated from some of the imported yarns used by the patient.
When was the last case of cutaneous anthrax?
The last case of cutaneous anthrax, before 2001, occurred in North Dakota, in 2000
Jim Young
Those who row the boat seldom have time to rock it.
A colleague (Hossler, F, East Tennessee State Med. Sch.) and I have used a method called corrosion casting to study the rich capillary bed underlying the trasitional epithelium of the urinary bladder. We used a modified commercial preparation "MERCOX Blue" for the preparation. The blue dye is, fortuitously, fluorescent. In a report given at the recent Philadelphia meeting of MSA (M&A, Vol6, Suppl 2, pp562-563), Czymmek et al. described the use of confocal microscopy in an analysis of corrosion casts. They used a Zeiss LSM 510 confocal (exciting the dye with the HeNe(Red) laser (633nm)) and detecting via a 660nm longpass emission filter. I have repeated this on an Olympus FV-300 using both the HeNe(R) as above with casts of pig bladder wall prepared by one of the above authors (Hossler). I have a particularly useful z-projection of a large, low power, image stack that I would be willing to send to requestors (69k, jpg) as an example of what can be accomplished with the application of confocal microscopy applied to low-power microanatomic studies of fluorescent objects. I have recommended the use of Evans Blue (C.I. 23860, CAS # 314-13-6) on several occasions in the past but have seen no results. This dye is known to form covalent bonds with albumin stoichimetrically and to be fluorescent in THAT form. It has been used in studies of extravasation around injury sites as well as for vascular volume studies. There is one reported incident of complicity in animal carcinogenesis (bolus, i.p. in rats) mentioned by IPCS INCHEM (http://www.inchem.org/documents/iarc/iarc/iarc121.htm).
Hope this is of some use.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
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} } I am developing a research proposal to investigate organic carbon } } colloids as a mechanism for contaminant transport of hydrophobic } } pollutants such as pesticides in surface and ground waters. The } } colloids generally range in size from 0.001 to 1 micron in size. I } } would like to characterize these organic colloids. I know that } } considering the low molecular weight of the carbon that this can be a } } problem for a lot of the microscopes and and complementary analytical } } devices. I was wondering if you had any suggestions for how I might } } characterize these organic colloids? } } } } Thank you for your help, } } Carol Creasey } } creasey-at-cats.ucsc.edu } } --
I have the chance of receiving a surplus PGT EDS detector. It has not been used for two years. My question is would the detector's window still be good? It has been in storage and the detector window has not been kept cold during these past two years. Any advice is appreciated.
Applications are invited for a full-time, non-tenure track position to operate a new electron microprobe/SEM facility in the Department of Geoscience at UNLV. The successful candidate will maintain a user-friendly facility for faculty, students, and visiting users, establish standard operating procedures (some to fulfill DOE quality assurance requirements), and solicit work from outside users. The candidate may conduct research as time permits; however, the primary focus of the position is to maintain the facility and to assist UNLV and outside users. The laboratory currently serves a wide range of research activity in geoscience, biology, chemistry, engineering, and materials science. Minimum requirements include a Master's degree in geology, geochemistry, or an appropriate related field, and electron microprobe and/or SEM experience. Review of applications will begin immediately and will continue until the position is filled. Salary will be commensurate with qualifications and experience. Position is contingent upon funding.
Interested applicants should send a CV, letter of interest detailing your interest in the position as described above, and names, addresses, phone numbers, and email addresses of at least three professional references to Dr. Jean Cline, Department of Geoscience, UNLV, 4505 Maryland Parkway, Box 454010, Las Vegas, NV 89154-4010; phone: 702 895 3262; fax: 702 895 4064, email: jcline-at-nevada.edu.
For information on the Geoscience department at UNLV visit our Web page at http://www.unlv.edu. UNLV is an Equal Opportunity/Affirmative Action employer. Persons are selected on the basis of ability without regard to race, color, sex, age, national origin, sexual orientation, religion, disability or veteran status.
Jean S. Cline Associate Professor, UNLV Department of Geoscience 4505 Maryland Parkway, Box 454010 Las Vegas, NV 89154-4010
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"Histochemistry" by Richard Horobin.
"Histological and Histochemical Methods" by John Kiernan
Most (all?) editions of "Cell and Tissue Biology" by Leon Weiss (editor) have a nice chapter on histochemistry by Helen Padykula.
Grizzi Fabio ICH wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear All } } I would like to receive any information about review papers on the basic } principles of histochemical techniques. I am quite interested to receive } references of papers on the basic characteristics of a histochemical method } (reproducibility, specificity etc). } } Many thanks. } } Fabio Grizzi } Scientific Direction } Istituto Clinico HUMANITAS } Milan, Italy
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
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Reimar,
It sounds to me very much like a rotifer: these are very common, and can survive considerable desiccation, and so are often found in roof gutters (where I remember finding them). I would guess that the "satellite dish" is the ciliated "wheel" from which the phylum gets its name - I think that the cilia are either moving too fast or simply too small to be seen at whatever magnification you are using. This generates a current, and I remember as a boy watching green spherical algae being sucked in and macerated.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
----- Original Message ----- } From: {reimar_gaertner-at-wsib.on.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, November 21, 2001 3:34 AM
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (elir-at-chem.ch.huji.ac.il) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, November 18, 2001 at 13:55:49 ---------------------------------------------------------------------------
Email: elir-at-chem.ch.huji.ac.il Name: Eli rothenberg
Organization: Hebrew University
Education: Graduate College
Location: Jerusalem, Israel
Question: I would like to know what is the correct setup for a wide illumination in a confocal microscopy arrangement, that is, what optics should i use in the path of the laser, if i want to get the maximum illuminated area through my objective, the setting is as follows: laser-collimated beam- dichroic mirror-objective-illuminated surface. (p.s I thought of using a short focusing lens that will focus half way through the objective so maybe that will give me a wide illumination on the surface, and/or maybe a diffuser to go along with it?)
I apologize if this message has been posted already. I responded with this information several days ago and it did not come back to my Inbox as a message distributed to the List. I wasn't sure if it had made it out or not.
Ken:
I don't have experience with either of the two scanners you mentioned. For our customers, we usually recommend Microtek. We have had good success with the (previously discussed here) ArtixScan 2500 which is 36-bit with a 3.4 dynamic range. Resolution is 2500 dpi (optical) and the list price is $4,499. (Usually available for less.) (10,000 element CCD)
Microtek also offers (but I have yet to use) the ArtixScan 1100 which is closer to the ones you mentioned. Optical resolution is 2000 x 1000, 42-bit, and a higher reported dynamic range of 3.9. This one has a list price of $1,499. (8,000 element CCD)
Both of these use dual plate technology including Microtek's patented Emulsion Direct Imaging Technology for incredibly sharp transparency scanning. With E.D.I.T., there is no glass between the scanning lens, film, or light source, eliminating distortion and Newton rings that occur with traditional transparency adapters.
I hope this helps. If I can be of any additional assistance, please let me know.
*Kind Regards, *Dave Hall *Resolution Technology, Inc - (614) 921-0045
-----Original Message----- } From: Ken Bart [mailto:kbart-at-hamilton.edu] Sent: Monday, November 26, 2001 10:09 AM To: microscopy-at-sparc5.microscopy.com
Hi: I am looking to replace my old Epson flat bed scanner and want a high quality scanner. This instrument is used primarily for scanning TEM negatives, 35 mm slides, some OCR and needs to be function in a multi-user environment. I have been considering the UMAX Powerlook 3000 and the Heidelberg Linoscan 2650 due to their reported 3.6 dynamic range and 42 bit pixel depth. Does anyone have experience with these machines? Is there something better?
Thanks!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
Has anyone successfully processesed tissue (in our case part of a rat brain) for TEM just after studying brain circulation by fluorescence microscopy? We use rhodamine and fluorescein tagged AB 40 and the experiment runs for about 1 1/2 hours. At that point, I fix the tissue with glutaraldehyde and osmium and embed it in epon-araldite. The problem is that membranes disappear. I'd been warned that would happen and it did. Is there any way to protect the membranes or any other stain or method that might help?
Thanks for any help! Margie Bryant Anatomy Dept. Univ. of South Florida Medical School 12901 Bruce D. Downs Blvd Tampa, Fl. 33612 email address: mbryant-at-com1.med.usf.edu
Dear Listers, Consolidation of our EM facilities has resulted in a surplussing a 1984 Philips 410 LS. It has been factory maintained and remains in excellent condition. The department is asking $30,000, or best offer. For more information or sales inquiry, please contact: Dr. Lesnick Westrum Dept. Neurological Surgery University of Washington (206) 543-5434 westrum-at-u.washington.edu
Regards, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax ************************************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
I heard on the radio that the economy is in a recession, so I figured I had better get busy and try to save money and recycle some supplies. I also had a big box of used SEM stubs and thought I could start there.
The stubs are left over from various projects, some have silver paint, some with double stick tape, etc. I thought I could loosen things up a bit by soaking them in water and/or an Alconox solution. This fit in very well with my plan for recycling since the plan was to let them soak until the recession is over.
Since there was no way to know when things would return to normal, I put some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. Says it is safe for aluminum and other things.
Now I have a problem. The stubs turned pretty ugly. Big black stains all over the place. Looks like some reaction between the Al and the Alconox. Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the black stuff. I can polish off the black stuff, but that wasn't part of the plan. This was supposed to be simple.
I don't think the black stuff affects the functioning of the stub, it just looks ugly and means I have to explain to some users that they are just as good as the few shiny ones left in the drawer (stubs that is, not users).
So tell me oh mighty metal and materials experts, what is the black stuff, why did it appear, and is there any easy way to get rid of it? Or is this some kind of plan to pull the economy back up to speed by making me buy a bunch of shiny new SEM stubs?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I need practical advice for a colleague on conjugating fluorophores to primary antibodies for immunofluorescence microscopy. She would like to use Alexa and/or Cyanine labels. Are there particular products/vendors that people have used with success? What are the problems and pitfalls to avoid?
Thanks in advance!
Dr. David P. Bazett-Jones Senior Scientist Programme in Cell Biology, Research Institute The Hospital for Sick Children 555 University Avenue Toronto, ON M5G 1X8 Canada
You will want to check if the window is OK, but the fact that the detector has been in storage warm for 2 years does not make it likely that it is broken.
If I had the chance (and needed the detector, of course!), I would accept it. I would pump it (for which you will need a pumping adapter) and see if you get a vacuum (do this *BEFORE* you put liq. N2 in it). You can get by with a rotary pump vacuum, but if you can get a high vacuum (we have a port plumbed into one of our evaporaters, with a valve, so we can pump dtectors), so much the better. If it pumps down, let it pump for a few hours, fill with nitrogen, then plug the pumping port and remove the adapter. Let it cool overnight, then fire it up. If you have access to a Fe55 radioactive source you won't even need to put it on the microscope to check it out, but I guess that you probably don't have one of those!
I have a pumping adapter for very old Kevex detectors with US threads on the plug - more recent detectors, and all those from Link have metric threads. I don't know about PGT, but if yours has US threads, you are welcome to borrow my adapter - I don't think they vary from manufacturer to manufacturer.
Good luck!
Tony Garratt-Reed
At 01:16 PM 11/29/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our experience is that 0.3 mm is better. 0.5 mm will work, but the width of the hologram for a given fringe spacing will be reduced. It depends on what you mean by "normal holography"...
Larry
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-- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I would ask advise concerning cover glass thickness specs. The objectives on my microscope are specified for .17 mm thick cover slips. I understand this is quite important.
I guess the answer must be obvious, but given that the idea mounting mediun would have the same refractive index as the cover slip, and I think the oil (in case of an immersion situation), how should one deal with this? the objects should be in direct contact with the cover slip?
Obviously this is not possible in all cases. Should one such as myself (unfortunate to have objectives with no correction collar) use even thinner cover slips to compensate for any distance between the object and the bottom side of the cover slip?
If I might pose another perhaps equally obvious question, is the reasoning for use of hanging drop preparations also the need to keep the specimen at this exact distance?
Then what does water do when doing wet mounts?
Sorry for wasting bandwidth on such ridiculous questions,
Alan Davis
-- Alan E. Davis, Science Instructor Marianas High School PMB 30, Box 10006, Saipan, MP 96950 Northern Mariana Islands adavis-at-saipan.com
"An inviscid theory of flow renders the screw useless, but the need for one non-existent." ---Lord Raleigh(aka John William Strutt),or else his son, Jr., who was also a scientist.
We have a collaborator who is having problems using colloidal gold-lectins for the post-embedding labeling of protozoa (lots of non-specific binding, even to Epon and Lowicryl embedding media). Does anyone have any good protocols for the use of these conjugates?
Thanks in advance,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
I don't know how you could get rid of those ugly stains, but you can probably get your clients to accept the blackened stubs without a murmur by saying that you "antiqued" then after seeing Martha Stewart do it :-)
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, November 29, 2001 8:34 PM
I have seen black "smut" on Al alloys etched with alkalis. Although I never analyzed it, I would guess it was undesolved Al alloying elements (Fe, Cu, etc). A quick dip in a medium strength nitric acid solution, followed by water rinse cleaned it right off.
Woody
} } I heard on the radio that the economy is in a recession, so I } figured I had } better get busy and try to save money and recycle some } supplies. I also had } a big box of used SEM stubs and thought I could start there. } } The stubs are left over from various projects, some have } silver paint, some } with double stick tape, etc. I thought I could loosen things } up a bit by } soaking them in water and/or an Alconox solution. This fit in } very well } with my plan for recycling since the plan was to let them } soak until the } recession is over. } } Since there was no way to know when things would return to } normal, I put } some into an ultrasonic cleaner with Fisher Ultrasonic } Cleaning solution. } Says it is safe for aluminum and other things. } } Now I have a problem. The stubs turned pretty ugly. Big black } stains all } over the place. Looks like some reaction between the Al and } the Alconox. } Stubs in plain water are not so bad. Ultrasonic cleaner can't } remove the } black stuff. I can polish off the black stuff, but that } wasn't part of the } plan. This was supposed to be simple. } } I don't think the black stuff affects the functioning of the } stub, it just } looks ugly and means I have to explain to some users that } they are just as } good as the few shiny ones left in the drawer (stubs that is, } not users). } } So tell me oh mighty metal and materials experts, what is the } black stuff, } why did it appear, and is there any easy way to get rid of } it? Or is this } some kind of plan to pull the economy back up to speed by } making me buy a } bunch of shiny new SEM stubs? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
For those of us with "text-only" email, Larry means 0.3 micrometers.
At 11:16 PM -0500 11/29/01, Larry Allard wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Shu-You: } } Our experience is that 0.3 mm is better. 0.5 mm will work, but the } width of the hologram for a given fringe spacing will be reduced. } It depends on what you mean by "normal holography"... } } Larry } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have used the Alexa kits from Mol Probes to conjugate their visible fluors to protein-G-purified IgG. I have always gotten satisfactory results, although my degree-of-labeling was always less than optimal as reported by Mol Probes. We do almost exclusively laser confocal microscopy, and I can say that the primaries I made are very useful with that approach. The visible Alexa fluors are very bright and very resistant to bleaching. I cannot comment on how they would work in standard epifluorescence, but my guess is that if one has a decent camera (i.e., short exposure times), they should work equally well for this approach. The only practical pitfall in the conjugation process is separation of unbound dye from conjugate. Mol Probes includes a simple gravity-run molecular sizing column in the kit with which to do this. However, if efficiency of labeling is low, it can be difficult to see the band you want and, as with any such separation approach, you will definitely experience some loss of material regardless of how critically you run the column. Also, if you have never set up a column before, this resin is very "gloply" and not trivial to pack. So, I thought, "why not dialyze away the unbound dye?" and Mol Probes says you can do this (they supply the column because you get faster results and they can justify a higher cost for the kits...). As I am never in a huge hurry, I tried this. As far as I could see, there was NO difference in the color intensity of the dialysed material! This suggests that I had 100% labeling (!). The upside on that particular prep was that, as Mol Probes also points out, if your immunofluorescence sample prep is good then you really don't have to worry a lot about unbound dye as it will be washed out any (only the conjugate should stick). This was the case for me with that dialyzed prep.
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Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
Commercially supplied stubs are anodized, which makes them more resistant to oxidizing, but with detergent that barrier breaks down. The dark stuff has to be aluminium oxide, which is much more stable then plain Al. You could argue that those mounts have character and that the appearance does not matter.
I used to recycle a percentage of mounts, but solvents and detergent are just too messy. I prefer a fine flat file (could also use sanding paper, a medium to coarse grade). Peel off any easily removed sticky material and then give the surface of the mount a few strokes on the file. A double layer of paper towel under the file collects most of the rubbish and protects the bench.
Its an ok job, if you don't expect to do that full-time for the rest of your working life. If you are entrepreneurial, leave the set-up with some old mounts on the bench and hide the new mounts. This is doubly effective if some influential people come to contemplate your funding problems while cleaning dirty mounts.
With a bit of tinkering you could have your lab's economy run counter-cyclical to the one Alan Greenspan is worried about. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, November 30, 2001 10:35 AM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I heard on the radio that the economy is in a recession, so I figured I had } better get busy and try to save money and recycle some supplies. I also had } a big box of used SEM stubs and thought I could start there. } } The stubs are left over from various projects, some have silver paint, some } with double stick tape, etc. I thought I could loosen things up a bit by } soaking them in water and/or an Alconox solution. This fit in very well } with my plan for recycling since the plan was to let them soak until the } recession is over. } } Since there was no way to know when things would return to normal, I put } some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. } Says it is safe for aluminum and other things. } } Now I have a problem. The stubs turned pretty ugly. Big black stains all } over the place. Looks like some reaction between the Al and the Alconox. } Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the } black stuff. I can polish off the black stuff, but that wasn't part of the } plan. This was supposed to be simple. } } I don't think the black stuff affects the functioning of the stub, it just } looks ugly and means I have to explain to some users that they are just as } good as the few shiny ones left in the drawer (stubs that is, not users). } } So tell me oh mighty metal and materials experts, what is the black stuff, } why did it appear, and is there any easy way to get rid of it? Or is this } some kind of plan to pull the economy back up to speed by making me buy a } bunch of shiny new SEM stubs? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu }
I have been asked to prepare some cross sections of hair for inspection with an optical microscope and not having much experience doing this I have gotten some advice locally and tried a few things with an old microtome, but without much success.
I am wondering if someone on this list could offer me some advice on how to prepare the hair for the sectioning. So far working with paraffin has not be successful as the hair seems to not cut well leaving either a hole in the wax strip or a raised and irregular hair.
Alternately I would be interested in talking to a lab near Connecticut that could do this sectioning for me.
Thanks very much for any help you may be able to offer!
Richard Shalvoy Arch Chemicals Cheshire, CT 203-271-4394 rbshalvoy-at-archchemicals.com
} } } I need practical advice for a colleague on conjugating fluorophores to } primary antibodies for immunofluorescence microscopy. She would like to } use Alexa and/or Cyanine labels. Are there particular products/vendors } that people have used with success? What are the problems and pitfalls } to avoid? } } Thanks in advance! } } Dr. David P. Bazett-Jones **************** Dear David, The Alexa family of dyes are sold by Molecular Probes, and if you buy them uncongugated, they come with instructions for congugating them to your Ab. I haven't done it personally, but my coworkers have and tell me its easy.
I have not financial interest in Molecular Probes, I'm just a satisfied customer. Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Jonathan Krupp wrote: ============================================================= I heard on the radio that the economy is in a recession, so I figured I had better get busy and try to save money and recycle some supplies. I also had a big box of used SEM stubs and thought I could start there.
The stubs are left over from various projects, some have silver paint, some with double stick tape, etc. I thought I could loosen things up a bit by soaking them in water and/or an Alconox solution. This fit in very well with my plan for recycling since the plan was to let them soak until the recession is over.
Since there was no way to know when things would return to normal, I put some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. Says it is safe for aluminum and other things.
Now I have a problem. The stubs turned pretty ugly. Big black stains all over the place. Looks like some reaction between the Al and the Alconox. Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the black stuff. I can polish off the black stuff, but that wasn't part of the plan. This was supposed to be simple.
I don't think the black stuff affects the functioning of the stub, it just looks ugly and means I have to explain to some users that they are just as good as the few shiny ones left in the drawer (stubs that is, not users).
So tell me oh mighty metal and materials experts, what is the black stuff, why did it appear, and is there any easy way to get rid of it? Or is this some kind of plan to pull the economy back up to speed by making me buy a bunch of shiny new SEM stubs? =================================================================== In the typical laboratory that "accummulates" no-longer-needed SEM mounts, the "history" of what was originally there on the mounts (e.g. the samples) is often times lost with the passing of the months if not also from the natural turnover of both employees and/or students. We have ourselves considered offering some kind of a "recycling" program for SEM mounts, but the inability to really know what we would be getting would pose to our employees an unacceptable safety risk. Furthermore, from the standpoint of shipping, some might argue that phenomenologially, "waste" mounts are akin in lots of ways to a hazardous waste, which would impose restrictions on how they could be shipped (if indeed they could be shipped easily at all).
Also, some laboratories have a way of accidentally or inadvertently "co- mingling" or mixing up used beryllium mounts with aluminum mounts, and once a part of the kind of "mess" that was described, they look no different from orginary aluminum mounts. Hence you do want to be further cautious when talking about polishing down or grinding, because you absolutely don't want to get into a beryllium dust exposure situation, which can surely occur if the polishing pads or grinding papers dry out.
Disclaimer: SPI Supplies offers new aluminum mounts for all SEMs, and we would like to see more new mounts being used. But we offer the above comment not to encourage the purchase of more new mounts, but only to point out that if you did want to "recycle" used SEM mounts, you do want to be careful not to mix up beryllium mounts with the aluminum and you also want to have some awareness of what kinds of samples, might be on the no-longer- needed mounts.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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"David P. Bazett-Jones" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I need practical advice for a colleague on conjugating fluorophores to } primary antibodies for immunofluorescence microscopy. She would like to } use Alexa and/or Cyanine labels. Are there particular products/vendors } that people have used with success? What are the problems and pitfalls } to avoid? } } Thanks in advance! } } Dr. David P. Bazett-Jones } Senior Scientist } Programme in Cell Biology, Research Institute } The Hospital for Sick Children } 555 University Avenue } Toronto, ON M5G 1X8 } Canada } } TEL: 416 813-2181 } FAX: 416 813-2235 } EMail: david.bazett-jones-at-sickkids.ca
Hi Alan, First, the mountant's RI is matched to that of the specimen. That of the immersion oil to that of the glass, although I cheat when I use Cargille oils of non-standard RI to view some difficult coverglassed specimens in order to alter the phase dynamics of the system. Second, the 0.17mm coverglass represents the average thickness of the No 1 coverglass that one normally uses for the purpose of mounting specimens for microscopy. Further, the glass components of the objective have been ground in expectation of that thickness of glass between the objective and the specimen, whether the objective is immersive or not. Even though the use of oil immersion to view a blood smear without a coverglass is common, the oil immersion objective almost always 'says' that it expected a coverglass to be there. Third, with respect to specimen thickness, there is nothing that can change the built-in working parameters of the objectives, thus, you won't be able to use oil immersion to any good effect on a whole mount of any specimen much thicker than 10um. A 40x objective is stressed beyond its limits when one tries to view a whole mounted 24-hr chick embryo. In other words, the working distance, which is not usually given, defines the limit of the distance between the face of the objective and the object one wishes to view. If one places physical impediments between face and specimen that exceed the working distance, then the specimen cannot be viewed by the scientist and is crushed by the freshman biology student who doesn't know about it. On the other side of that is the fact that were the working distance of the oil immersion lens too long, the oil might not be able to bridge the gap between objective and coverglass. Fourth, the information on the barrel of an objective is part of the description of the optical properties of the objective. Its design parameter set, if you will. The 0.17 correction tells much about working distance. The numerical aperture tells a lot about the optical components used inside the objective. Fifth, I always thought that the hanging drop was to provide a method for viewing live material in a confined environmentally friendly volume (cage).
} ---------- } From: Alan Davis } Sent: Thursday, November 29, 2001 5:27 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: cover glass thickness } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would ask advise concerning cover glass thickness specs. The objectives } on my microscope are specified for .17 mm thick cover slips. I understand } this is quite important. } } I guess the answer must be obvious, but given that the idea mounting } mediun would have the same refractive index as the cover slip, and I } think the oil (in case of an immersion situation), how should one deal } with this? the objects should be in direct contact with the cover slip? } } Obviously this is not possible in all cases. Should one such as myself } (unfortunate to have objectives with no correction collar) use even } thinner cover slips to compensate for any distance between the object and } the bottom side of the cover slip? } } If I might pose another perhaps equally obvious question, is the reasoning } for use of hanging drop preparations also the need to keep the specimen at } this exact distance? } } Then what does water do when doing wet mounts? } } Sorry for wasting bandwidth on such ridiculous questions, } } Alan Davis } } -- } Alan E. Davis, Science Instructor } Marianas High School } PMB 30, Box 10006, } Saipan, MP 96950 } Northern Mariana Islands } adavis-at-saipan.com } } } "An inviscid theory of flow renders the screw useless, but the need } for one non-existent." ---Lord Raleigh(aka John William Strutt),or } else } his son, Jr., who was also a scientist. } }
} Commercially supplied stubs are anodized, which makes them } more resistant to } oxidizing
It is my understanding that anodized aluminum has much thicker oxide layer than a natural oxide film, and, as a result, it should lead to the coloring of the metal (which we do not see for the stubs). Im I wrong?
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
The old AO Spencer rotary microtome manual recommends Pike Oil and a Neutral grease. I haven't used Pike oil, but I usually purchase a light machine oil. For use in colder environments such as when used for cryotomy, this manual recommended a lighter Hamilton watch oil. Clock oil will do. Others have suggested Singer sewing machine oil. I have used "Bear Oil" from Norton ("Formerly Pike Oil") which now appears to be Norton Part # 07660 787940 0, 4.5oz "Sharpening Oil" [URL: http://www.nortonconsumer.com/catalog/displaytier.asp?tier_id=D110050050010& display=all].
Hope this helps. Getting the proper lubricating oil is always a problem for these old machines.
P.S.1 I just purchased a new volume of "3 to 1" oil, and was surprised at its viscosity which I consider to be higher than in previous purchases.
P.S.2 A good rule is that if the oil hole contains a wick, the oil should be really light! Whatever, the lubricant should NOT be so light that it leaks out of the bearing.
Regards to all,
Fred Monson
} ---------- } From: Jonathan Wilson } Reply To: Jonathan Wilson } Sent: Friday, November 16, 2001 4:51 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: microtome oil } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I have been loaned a microtome (Leitz 1512) by a colleague which } according to the instruction manual requires regular oiling. The microtome } did not come with any oil and the price I have been quoted by a local } supplier for microtome oil was about 75$US (including VAT) for 50ml } (SAKURA } 1447). I found the price a bit steep and am left wondering if it is } absolutely necessary to buy "microtome" oil or if I can use an off the } shelf } oil (e.g 3 in 1 or something like that). I do not want to compromise the } performance of the microtome as it is not mine and would appreciate any } advice from those with experience. } Thanks for any help. } Sincerely, } Jonathan } } Jonathan Wilson (PhD) } CIIMAR } Rua do Campo Alegre 823 } 4150-180 Porto, Portugal } tel: 351 22 608 0470 / 71 } fax: 351 22 606 0423 } e-mail: wilson_jm-at-cimar.org } web: www.cimar.org } } }
I can't put cute subject headings anymore, so I'm trying not to be too negative ;-).
I'm working in a lab where they have been adding a hypo clearing agent called Orbit Bath into the Kodak rapid fix. Has anybody heard of doing this? I have never heard of this and I can't get the Orbit Bath without promising my first born and pulling a few teeth.
I'd rather go back to the tried and true method of developing the Kodak 4489 in HRP or D-19, fixing in Kodak Rapid Fix (using A & B), dunking in Hypo clear and washing as usual.
If someone can come up with a good reason to keep continuing this practice, or even an explanation as to why they even started it, I'm all ears.
Help clear me of my negative thoughts,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
No. 1 coverslips have a range of 0.13 to 0.17 (for Fisher) or 0.13 to 0.16 (for Corning brand).
No. 1 1/2 range from 0.16 to 0.19 for both Corning and Fisher.
Having used a micrometer on both sizes and brands, I find the most 0.17 coverslips in No. 1 1/2 boxes.
Tom
} } } Hi Alan, } First, the mountant's RI is matched to that of the specimen. That } of the immersion oil to that of the glass, although I cheat when I use } Cargille oils of non-standard RI to view some difficult coverglassed } specimens in order to alter the phase dynamics of the system. } Second, the 0.17mm coverglass represents the average thickness of } the No 1 coverglass that one normally uses for the purpose of mounting } specimens for microscopy. Further, the glass components of the objective } have been ground in expectation of that thickness of glass between the } objective and the specimen, whether the objective is immersive or not. Even } though the use of oil immersion to view a blood smear without a coverglass } is common, the oil immersion objective almost always 'says' that it expected } a coverglass to be there. } Third, with respect to specimen thickness, there is nothing that can } change the built-in working parameters of the objectives, thus, you won't be } able to use oil immersion to any good effect on a whole mount of any } specimen much thicker than 10um. A 40x objective is stressed beyond its } limits when one tries to view a whole mounted 24-hr chick embryo. In other } words, the working distance, which is not usually given, defines the limit } of the distance between the face of the objective and the object one wishes } to view. If one places physical impediments between face and specimen that } exceed the working distance, then the specimen cannot be viewed by the } scientist and is crushed by the freshman biology student who doesn't know } about it. On the other side of that is the fact that were the working } distance of the oil immersion lens too long, the oil might not be able to } bridge the gap between objective and coverglass. } Fourth, the information on the barrel of an objective is part of the } description of the optical properties of the objective. Its design } parameter set, if you will. The 0.17 correction tells much about working } distance. The numerical aperture tells a lot about the optical components } used inside the objective. } Fifth, I always thought that the hanging drop was to provide a } method for viewing live material in a confined environmentally friendly } volume (cage). } } } ---------- } } From: Alan Davis } } Sent: Thursday, November 29, 2001 5:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: cover glass thickness } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would ask advise concerning cover glass thickness specs. The objectives } } on my microscope are specified for .17 mm thick cover slips. I understand } } this is quite important. } } } } I guess the answer must be obvious, but given that the idea mounting } } mediun would have the same refractive index as the cover slip, and I } } think the oil (in case of an immersion situation), how should one deal } } with this? the objects should be in direct contact with the cover slip? } } } } Obviously this is not possible in all cases. Should one such as myself } } (unfortunate to have objectives with no correction collar) use even } } thinner cover slips to compensate for any distance between the object and } } the bottom side of the cover slip? } } } } If I might pose another perhaps equally obvious question, is the reasoning } } for use of hanging drop preparations also the need to keep the specimen at } } this exact distance? } } } } Then what does water do when doing wet mounts? } } } } Sorry for wasting bandwidth on such ridiculous questions, } } } } Alan Davis } } } } -- } } Alan E. Davis, Science Instructor } } Marianas High School } } PMB 30, Box 10006, } } Saipan, MP 96950 } } Northern Mariana Islands } } adavis-at-saipan.com } } } } } } "An inviscid theory of flow renders the screw useless, but the need } } for one non-existent." ---Lord Raleigh(aka John William Strutt),or } } else } } his son, Jr., who was also a scientist. } } } }
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
The thickness of a natural alumina film is in the order of some tens of nanometers. An anodized layer, on the other hand, can reach the micrometer level. However, it is porous. Usually, we can fill those pores simply by immersing parts in boiling or hot water. The film is nearly transparent or translucent. The color we see is actually the color of the dye used during pore filling for decoration purpose. There are many colors we can choose.
Kai Lorcharoensery
Materials Science & Engineering Lehigh University Bethlehem, PA
It is my understanding that anodized aluminum has much thicker oxide layer than a natural oxide film, and, as a result, it should lead to the coloring of the metal (which we do not see for the stubs). Im I wrong?
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Anodizing of aluminum can be done in many colors, as well as a clear coating. I would not expect SEM stubs to be anodized, as the aluminum oxide coating which forms is an insulator. This would inhibit the removal of the electron charge from the sample. The native oxide film is no where near the thickness of the usual anodized film, and can easily be broken. The thickness of the anodized film can be controlled, but it is still an effective insulator. I am fairly certain that none of the stubs I have ever used were anodized.
} From: "Paula Sicurello" {patpxs-at-gwumc.edu} } } I'm working in a lab where they have been adding a hypo clearing agent called Orbit Bath into the Kodak rapid fix. Has anybody heard of doing this? I have never heard of this and I can't get the Orbit Bath without promising my first born and pulling a few teeth. } } I'd rather go back to the tried and true method of developing the Kodak 4489 in HRP or D-19, fixing in Kodak Rapid Fix (using A & B), dunking in Hypo clear and washing as usual. } } If someone can come up with a good reason to keep continuing this practice, or even an explanation as to why they even started it, I'm all ears. } Some people use Orbit bath in the fix to cut the time in the fixer. I know of no advantage to it except it speeds up the process a little but not much. The main advantage is for fixing fiber paper so the paper spends less time in the fix and there is less time for fixer to diffuse into the paper.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
I'm certain that every mount sold by ProSciTech is anodized, and by their very appearance, all other such commercially sold mounts I have seen are anodized too. True the anodized film is very thin. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, December 01, 2001 9:07 AM, Darrell Miles [SMTP:milesd-at-US.ibm.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Anodizing of aluminum can be done in many colors, as } well as a clear coating. I would not expect SEM stubs to be } anodized, as the aluminum oxide coating which forms is an } insulator. This would inhibit the removal of the electron } charge from the sample. The native oxide film is no where } near the thickness of the usual anodized film, and can easily } be broken. The thickness of the anodized film can be } controlled, but it is still an effective insulator. I am fairly } certain that none of the stubs I have ever used were anodized. } } Regards, } Darrell
Tom, The window should be OK IF: 1.) it was protected from mechanical damage (covered effectively) and
2.) the vacuum in the dewar had not deteriorated much while it was cold, ie there was still a vacuum after it warmed up.
The question really is: is the SiLi crystal still good? Upon warming up, it may have gotten contaminated by all of the junk coming out of the zeolite cry-sorption material.
Basically, many detectors do store fairly well, but I would not invest much, if any, money in aquiring one unless is was proved to be good. On the other hand, if you can get it for free, the mounting is correct for your microscope and you're willing to have it rebuilt, it could be a good deal. Contact
"Jim Nicolino" {JNicolino-at-xraydetectors.com}
for possible cost. He's done several rebuilds for some of my customers.
Ken Converse owner Quality Images third party SEM service Delta, PA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have the chance of receiving a surplus PGT EDS detector. It has not been } used for two years. My question is would the detector's window still be } good? It has been in storage and the detector window has not been kept } cold during these past two years. Any advice is appreciated. } } Tom Bargar } tbargar.unmc.edu } } } }
True, the most likely defect of a soft-vacuum, stored SiLi detector would be a contaminated crystal. This "may" be worth fixing in the USA. In Australia, with the added high freight cost, such detectors become ornaments. At one point I had three SiLi detectors in use and two attractive, high-technology vases with dried flower arrangements. Cheers Jim Darley ProSciTech
On Saturday, December 01, 2001 11:27 PM, Ken Converse [SMTP:qualityimages-at-netrax.net] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Tom, } The window should be OK IF: } 1.) it was protected from mechanical damage (covered effectively) and } } 2.) the vacuum in the dewar had not deteriorated much while it was } cold, ie there was still a vacuum after it warmed up. } } The question really is: is the SiLi crystal still good? Upon warming } up, it may have gotten contaminated by all of the junk coming out of } the zeolite cry-sorption material. } } Basically, many detectors do store fairly well, but I would not invest } much, if any, money in aquiring one unless is was proved to be good. On } the other hand, if you can get it for free, the mounting is correct for } your microscope and you're willing to have it rebuilt, it could be a } good deal. Contact } } "Jim Nicolino" {JNicolino-at-xraydetectors.com} } } for possible cost. He's done several rebuilds for some of my customers. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } } "tbargar-at-unmc.edu"-at-sparc5.microscopy.com wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I have the chance of receiving a surplus PGT EDS detector. It has not been } } used for two years. My question is would the detector's window still be } } good? It has been in storage and the detector window has not been kept } } cold during these past two years. Any advice is appreciated. } } } } Tom Bargar } } tbargar.unmc.edu } } } } } } } }
Do you need to observe the hair cross-sections in great detail, or are you just looking at general morphology? If the latter, then you can cross-section hairs for microscopy by pulling the hairs through a small hole (0.5 or so mm diameter) in a flat metal sheet (can't remember exactly, but maybe 1 mm thick - about the same size as a microscope slide). You will probably need to mix the hairs with cotton so they go through and then stay tight in the hole - they need to be jammed fairly hard in the hole for the next part. Then you just cut off the hairs either side with a sharp razor blade so you get a flat cross-section, mount the metal slide on top of a regular slide, apply a drop of oil (or preferred mountant) to the hairs in the hole, coverslip and observe.
This is one way I've seen zoologists identify hairs and characterise differences. } } I have been asked to prepare some cross sections of hair for inspection with } an optical microscope and not having much experience doing this I have } gotten some advice locally and tried a few things with an old microtome, but } without much success. } } I am wondering if someone on this list could offer me some advice on how to } prepare the hair for the sectioning. So far working with paraffin has not } be successful as the hair seems to not cut well leaving either a hole in the } wax strip or a raised and irregular hair. } } Alternately I would be interested in talking to a lab near Connecticut that } could do this sectioning for me. } } Thanks very much for any help you may be able to offer! } } Richard Shalvoy } Arch Chemicals } Cheshire, CT } 203-271-4394 } rbshalvoy-at-archchemicals.com
cheers, rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
I would certainly hope that SEM stubs are NOT anodized. Not only does anodizing create an insulating surface, but it also creates a surface with very poor vacuum properties -- anodized parts will pump down slowly due to the gas trapped in the porosity.
Fred Schamber ASPEX, LLC
Jim at Proscitech wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm certain that every mount sold by ProSciTech is anodized, and by their very } appearance, all other such commercially sold mounts I have seen are anodized } too. True the anodized film is very thin. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Saturday, December 01, 2001 9:07 AM, Darrell Miles [SMTP:milesd-at-US.ibm.com] } wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Anodizing of aluminum can be done in many colors, as } } well as a clear coating. I would not expect SEM stubs to be } } anodized, as the aluminum oxide coating which forms is an } } insulator. This would inhibit the removal of the electron } } charge from the sample. The native oxide film is no where } } near the thickness of the usual anodized film, and can easily } } be broken. The thickness of the anodized film can be } } controlled, but it is still an effective insulator. I am fairly } } certain that none of the stubs I have ever used were anodized. } } } } Regards, } } Darrell
I have question: Is it possible to attach EDS detector to old EM400 WITHOUT goniometer ? 1) What one need to change - polepieces to have a hole ? apertures for low bkgnd ? or maybe whole goniometer has to be installed ? 2) Does maybe someone have such detector for give/sell away ??
regards
Krzysztof Herman EO Service Labsoft, ul.Ba¿ancia 45A 02-892 Warszawa, tel/fx: (+48 22)6449753, 6449750 mobile: (+48 601)307456 www.labsoft.com.pl
I am interested in gaining insight into the role of microscopic artifacts in the history of biology. May I impose on list members to contribute particularly glaring examples of misinterpreation of biological facts due to improper microscopic technique? I apologize if this is off-topic or a waste of bandwidth.
Let me provide the first example of a misinterpretation and request your assistance in learning whether this was due to improper use of the microscope, or perhaps even malfeasance: Sidney Hickson's early work on Millepora spp. (Cnidaria:Hydrozoa) fire corals. It seems an incredible lapse, a wholly fabricated natural history account, one that persisted for a considerable period in the fabric of the mythology of biological knowledge. I am interested because reproduction of Millepora platyphylla is the subject of incompleted thesis research of mine.
Hickson published a report on reproduction of "Millepora" around the end of the 19th Century. (Anong his other errors he synonomyized all species of Millepora as ecomorphs of one, M. alcicornis.) In this report, which I do not have available at this time, he included several plates of drawings depicting a putative sequence of reproductive events in this organism. We now understand that his depiction is not even close to the way that Millepora spp. (which were later redesignated as proper individual species through painstaking work by Boschma---notwithstanding the issues recently raised by molecular work) reproduce. The depiction involved dozens of drawings, and a sequence of events based on a misinterpretation of what are apparently artifacts.
Hickson (of Cambridge University) worked extensively in the field, including Indonesia and the Philippines. Was his microscopic work done in the field? Are members of this list enlightened as to Hickson's methods?
Hickson's erroneous drawings of the medusae of Millepora lived on for over 3/4 of a century in virtually every Invertebrate Zoology textbook published until the late 1980s or 1990s. His erroneous description of the medusa of Millepora as lacking a velum led to the designation of a separate branch of hydromedusae by Mayer, as the only hydrozoan medusa without a velum. My unpublished observations in the 1980s as well as published observations by John Lewis of McGill University showed that the medusae of Millepora spp. clearly possess a velum.
I apologize for monopolizing the bandwidth. I hope this is as fascinating a topic for others as for myself, and not considered off-topic.
Alan Davis
-- Alan E. Davis, Science Instructor Marianas High School PMB 30, Box 10006, Saipan, MP 96950 Northern Mariana Islands adavis-at-saipan.com
"An inviscid theory of flow renders the screw useless, but the need for one non-existent." ---Lord Raleigh(aka John William Strutt),or else his son, Jr., who was also a scientist.
We wish to set up a film thickness monitor to satisfy the wants of a user who has to work to ISO 9000 rules. We have a used Varian Auto deposition system Model 985-7009 which has become separated from its manual and which has problems.
Does anyone out there have a copy which we could have/get a copy of?
Thanks. Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
You are hoping in vain, commercial mounts are anodized, because: 1 The layer is too thin to insulate even at low voltages. SEM have high volts (even specimen currents are many and not fractions of volts) and low currents. No problem. 2 Otherwise the mounts would oxidise and I believe that too is not the best conductor. 3 Al too is porous, if you really care use polished stainless. 4 Does it matter? It would in an ion pumped TEM, but in an SEM? 5 All other suppliers do anodize. That is a better reason than you may think! 6 If we did not, people would buy other supplier's mounts because ours would look "cheap"
Now, I have a question: How long have you and some other people been using anodized mounts and not realised that the nice shiny finish is not a polish. Have you never seen machine finished Al???
If you really want to make a point, place a hundred commercial mounts into the bottom of the SEM chamber and then repeat that experiment with machine finished Al mounts. Record vacuua at various points and run the experiment three times. Report the results. I would be interested to learn if there is any difference. Those hundred plain Al mounts made in a machine shop would be a trifle expensive (you could acid strip the anodizing), but cheaper than most experiments.
Christmas is coming, so lets be charitable: anodized mounts are a good deal and they are very suitable for SEM work. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Monday, December 03, 2001 12:14 PM, Frederick Schamber [SMTP:schamber-at-aspexllc.com] wrote: } I would certainly hope that SEM stubs are NOT anodized. Not only does } anodizing } create an insulating surface, but it also creates a surface with very poor } vacuum } properties -- anodized parts will pump down slowly due to the gas trapped in } the } porosity. } } Fred Schamber } ASPEX, LLC } } Jim at Proscitech wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } I'm certain that every mount sold by ProSciTech is anodized, and by their } } very } } appearance, all other such commercially sold mounts I have seen are } } anodized } } too. True the anodized film is very thin. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ABN: 99 724 136 560 www.proscitech.com } } } } On Saturday, December 01, 2001 9:07 AM, Darrell Miles } } [SMTP:milesd-at-US.ibm.com] } } wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Anodizing of aluminum can be done in many colors, as } } } well as a clear coating. I would not expect SEM stubs to be } } } anodized, as the aluminum oxide coating which forms is an } } } insulator. This would inhibit the removal of the electron } } } charge from the sample. The native oxide film is no where } } } near the thickness of the usual anodized film, and can easily } } } be broken. The thickness of the anodized film can be } } } controlled, but it is still an effective insulator. I am fairly } } } certain that none of the stubs I have ever used were anodized. } } } } } } Regards, } } } Darrell
I looking at purchasing a microtome for sectioning polymers, primarily PE. Although I am familiar with microtomes used in biological studies, I have no experience in sectioning polymers. Has anyone sectioned polymers with repeated success? What features are desired for sectioning polymers? Is there a specific knife angle that works well?
KD Derr SEM/Polymer Technician Gas Technology Institute 1700 S Mount Prospect Rd Des Plaines, IL 60018 (847) 768-0505 (phone) (847) 768-0569 (fax) kd.derr-at-gastechnology.org
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I have never used Orbit bath nor Hypo. In fact the former is totally new to me. I have heard of Hypo in the '60s. I was curious and went to the expert. I phoned Kodak Scientific Support (800-225-5352) to find out whether developing the 4489 film in D19 and fixing in Kodak Rapid Fixer is adequate. The answer is "Yes" What about Hypo? He asked, "Who makes it?" He assured me that nothing else is needed after Rapid Fix. Just wash the negs. in water will do.
AnnFook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Room 2091, Bldg. 20, Central Experimental Farm, Ottawa, Ontario Canada K1A 0C6
Tel: 1-613-759-1638 Fax: 1-613-759-1701
e-mail: yanga-at-em.agr.ca
} } } "Paula Sicurello" {patpxs-at-gwumc.edu} 11/30/01 03:25PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Listers,
I can't put cute subject headings anymore, so I'm trying not to be too negative ;-).
I'm working in a lab where they have been adding a hypo clearing agent called Orbit Bath into the Kodak rapid fix. Has anybody heard of doing this? I have never heard of this and I can't get the Orbit Bath without promising my first born and pulling a few teeth.
I'd rather go back to the tried and true method of developing the Kodak 4489 in HRP or D-19, fixing in Kodak Rapid Fix (using A & B), dunking in Hypo clear and washing as usual.
If someone can come up with a good reason to keep continuing this practice, or even an explanation as to why they even started it, I'm all ears.
Help clear me of my negative thoughts,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
To "Anodize" is to subject a metal to an electrolytic action as the anode of a cell in order to coat/deposit a protective or decorative film.
Most aluminium anodization that I have seen creates a dark THICK oxide film (tens to hundreds of nm thick) and is insulating.
I've never seen this on an aluminium SEM stub. Certainly many I've used are electropolished and some (the ones we make here in our machine shop ) are simply milled and mechanically polished. All will at a minimum have a native oxide. All are conductive, and yes (Jim) I've seen polished (electrochemical, mechanical) as well as all ranges of "machine" finished products. These are NOT anodized, they are all simply oxidized by leaving them sit in air. The native oxide which forms is a few tens of angstroms thick and is stable, by definition this is not anodization. I have never heard of anyone calling this process "anodization" should that be what you are referring to.
So... now I'm curious and I would request that the various suppliers who like reply to this list to fill us all in on what you do.
1.) How many of you anodize your stubs (ie. coat), vs electropolish, vs mechanically polish. 2.) If you intentionally anodize, (ie. deposit/coat) your stub with an electrodeposited layer how thick is your film, and what do you anodize it with. 3.) It might also help to define the "anodization" process (if you do this) so that we are all understand what each of you mean in the context of how you make the stubs we purchase.
Orbit bath is very useful. It should not, however, be added to the fixer. The purpose of the hypo clearing agent is to remove the residue of fixer from the negatives after fixing. I strongly recommend that Orbit not be added to the fixing bath.
Good luck,
Gary M. Brown
Jim at Proscitech To: "'Darrell Miles'" {milesd-at-US.ibm.com} , {jim-at-proscitech "Microscopy-at-sparc5.microscopy.com" .com} {Microscopy-at-sparc5.microscopy.com} cc: Subject: RE: Cleaning SEM stubs 12/01/01 05:16 AM Please respond to "jim-at-proscitech .com"
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I'm certain that every mount sold by ProSciTech is anodized, and by their very appearance, all other such commercially sold mounts I have seen are anodized too. True the anodized film is very thin. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, December 01, 2001 9:07 AM, Darrell Miles [SMTP:milesd-at-US.ibm.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Anodizing of aluminum can be done in many colors, as } well as a clear coating. I would not expect SEM stubs to be } anodized, as the aluminum oxide coating which forms is an } insulator. This would inhibit the removal of the electron } charge from the sample. The native oxide film is no where } near the thickness of the usual anodized film, and can easily } be broken. The thickness of the anodized film can be } controlled, but it is still an effective insulator. I am fairly } certain that none of the stubs I have ever used were anodized. } } Regards, } Darrell
Orbit Bath was, as you say, a hypo-clearing agent designed to decrease water wash times while increasing removal of residual fixer from photographic materials after processing. I believe it is no longer made. We recently sent a BUNCH of this stuff to our chemical recycling folks here on campus, because we do so little silver-based photographic work anymore. We retained a small stock.
Although also used for films, Orbit and similar chemicals had their heydey before the routine use of resin-coated papers, which don't absorb chemicals readily. The fiber-based materials used commonly in the past were (and are) very elegant papers and for my money are still the best photographic papers made. However, for anything other than fine-arts use, resin coated papers have rightfully taken over, with the advantages of decreased processing times and chemical consumption.
By way of interesting trivia, I had once heard that some of the fixer-removal products got their start after someone discovered that washing photo materials in seawater after processing (maybe because it was the only water they had available in an emergency) was more efficient than fresh water. Does anyone know if this has a basis in fact or is it just urban myth?
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Paula Sicurello [mailto:patpxs-at-gwumc.edu] Sent: Friday, November 30, 2001 2:26 PM To: microscopy-at-sparc5.microscopy.com
Hi Listers,
I can't put cute subject headings anymore, so I'm trying not to be too negative ;-).
I'm working in a lab where they have been adding a hypo clearing agent called Orbit Bath into the Kodak rapid fix. Has anybody heard of doing this? I have never heard of this and I can't get the Orbit Bath without promising my first born and pulling a few teeth.
I'd rather go back to the tried and true method of developing the Kodak 4489 in HRP or D-19, fixing in Kodak Rapid Fix (using A & B), dunking in Hypo clear and washing as usual.
If someone can come up with a good reason to keep continuing this practice, or even an explanation as to why they even started it, I'm all ears.
Help clear me of my negative thoughts,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We have been using carbon grating replicas for calibrating the magnification of our TEM images for many years. We are in the process of revising our imaging and morphometry protocols. I am not happy with the precision and longevity of the carbon grating replicas. Are there more precise and/or stronger standards for TEM magnification calibration? The final magnifications of our images range from 3,000 -20,000 X.
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-umn.edu
Fred Schamber wrote: =============================================================== I would certainly hope that SEM stubs are NOT anodized. Not only does anodizing create an insulating surface, but it also creates a surface with very poor vacuum properties -- anodized parts will pump down slowly due to the gas trapped in the porosity. ================================================================ Fred is certainly correct.
SPI Supplies has offered "lathe finish" aluminum SEM mounts for nearly thirty years and while we have offered different types of surface finishes, none have ever involved the use of an anodization process. I think the same would be true for the other major manufactures of SEM mounts but I would not want to speak for them since I really don't know exactly what they might or might not have offered over the years. To apply a nonconductive layer to an SEM mount would surely, to me at least, seem quite counter productive.
I think that there is a problem with semantics here. Let me quote from my trusty Webster's New Word Dictionary:
----------------------- an•o•dize to put a protective, often colored, oxide film on (a light metal) by an electrolytic process in which the metal serves as an anode. ----------------------
I was happy to see that my memory was not failing me.
The nornal, naturally occurring oxide layer that forms on bare exposed aluminum is just that: an oxide layer. It is very thin, and generally not enough to cause anyone real complications with charging. If this is the layer people are talking about, then so far as I know, no one (well almost no one) would consider this to be an "anodized" layer. After all, we are talking about a layer intentionally applied, not one that forms naturally upon exposure to air.
I can not comment about what some other firm(s) might be doing, but if they are indeed anodizing their aluminum mounts, they would be growing a nice oxide layer, much thicker than the naturally occurring kind, and one that certainly would exhibit lower conductivity characteristics.
SPI Supplies and to the best of my knowledge, the other leading firms offering SEM mounts, offer aluminum (often times an alloy rather than pure aluminum) that is either "lathe finish", or "polished" or an intermediate finish, called by SPI Supplies as its Luster™ finish. But in no case is an oxide layer being intentionally grown onto the aluminum surface using any kind of anodization process. Details about the SEM mounts produced by SPI Supplies can be found on the SPI Supplies website given below.
Actually, what is involved here is a bit more than just semantics. The naturally formed oxide layer is many times thinner than an anodized layer, and if my memory is right, that naturally occurring layer is on the order of 5 nm or less. Most anodized layers we have seen over the years (by cross- section TEM, for example) are on the order of 500 - 1000 nm (or more), and all exhibit high porosity (and therefore surface area). Until now, I have not been aware that anyone would call the naturally formed oxide layer to be an anodized layer, but perhaps I am not keeping up with the more modern use of the term. Someone help me out if I am wrong about this.
Chuck
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I will be the first to admit that I am not an expert on either SEM stubs or anodizing, but I have had very negative experience with the latter in vacuum systems. I once designed some relatively small black-anodized parts into a SEM and spent considerable time trying to find out why we had a degradation in pumping speed -- finally, someone wiser than me sanded off the anodizing and the vacuum was fine. So I have, in effect, done the experiment you describe -- ordinary machined aluminum parts are vastly superior to anodized aluminum parts in terms of their vacuum properties in my experience (and I did some followup experiments in a test system employed specifically to measure vacuum properties of materials and confirmed that a little bit of anodizing is a surprisingly bad thing). If one actually did the experiment you propose with one hundred anodized stubs, my bet is that you would be shocked (but in order to do the experiment, I think you would first have to have MOST stubs anodized). It is not a question of the mere "roughness" of the surface.Apparently the anodizing process creates very tiny interstices which hold the dye, and this is what causes the problem -- or so I have assumed. I suppose it could be the dye itself -- I never tried the experiment on "clear anodized" parts.
When I look at catalog advertisements for aluminum SEM stubs, I always see reference to various degress of polishing, and don't recall ever seeing reference to anodizing. Reading the description, it certainly sounds like they are just pure uncoated aluminum -- I've never looked at the ones you sell.
} From an electrical standpoint, I also really wonder how anodizing could work. I know that anodizing must be avoided on electrical cabinetry because one cannot establish reliable contacts (a different surface treatment process is used on aluminum instead). I'm sceptical about the "thin coating" concept since it is my understanding that it is precisely the insulating layer which protects the surface from oxidation. Any amount of insulation on a SEM stub would be a big problem. (Of course, the same thing can be said about the thin oxide layer that naturally forms when aluminum is exposed to air.) But it is my understanding that anodizing is a process which modifies the surface, not a simple "coating" which can be "thin".
My negative opinions about anodizing for vacuum systems aren't just mine -- in fact, I thought they were "common knowledge". For example, I refer you to the very informative vacuum information site maintained by Roy Schmaus at the University of Alberta http://nyquist.ee.ualberta.ca/~schmaus/vacf/index.html Under Basics/An Introduction to Materials for use in Vacuum we find the following statement: "Aluminum that will be exposed to vacuum should never be anodized due to serious outgassing problems." Under "References" on the same page he notes: "I recently had a call from someone who has used Anodized Aluminum as an INSULATOR down into the ultra high vacuum region. Admittedly the pieces used were very small and INITIAL OUTGASSING WAS A PROBLEM but low costs were a major advantage." [my EMPHASIS added]
So given my own negative experience and the congruent remarks of others who work in this field, I have long understood that anodizing was something to always avoid in vacuum systems -- and I've ASSUMED that applied to SEM stubs. Am I wrong?
How DO polished stubs stay so shiny? I've wondered the same. However, anodized parts I have seen always have a kind of "matte" appearance, rather than a high gloss in any case.
So, though it is entirely possible that I will learn something from this discussion that I did not know previously (it would hardly be the first time), my limited grasp of the subject still convinces me that SEM stubs should NOT be anodized. But how do they retain their surface finish and conductivity? Your response suggests that you manufacture SEM stubs and call out an anodized surface -- did I read this correctly? Or are you purchasing stubs which you assume are anodized? I really would be interested if someone who is in the business of manufacturing these things could comment directly to this -- without disclosing proprietary information of course.
"Charity" in view of the season notwithstanding, I consider this a matter of considerable practical importance. If I am in error about this, I want to know it, since a non-insulating, non-outgassing, but still protective anodizing surface would be a very good thing. If I am correct, then others reading this exchange should know that anodizing is something that should be scrupulously avoided in vacuum systems (SEM and otherwise). This is the kind of thing where a little misinformation can cause a great deal of grief, so getting to the bottom of this will presumably be a good thing for everyone. I'm prepared to learn from new facts, if someone can put them on the table.
Fred Schamber ASPEX, LLC
Jim at Proscitech wrote:
} You are hoping in vain, commercial mounts are anodized, because: } 1 The layer is too thin to insulate even at low voltages. SEM have high volts } (even specimen currents are many and not fractions of volts) and low currents. } No problem. } 2 Otherwise the mounts would oxidise and I believe that too is not the best } conductor. } 3 Al too is porous, if you really care use polished stainless. } 4 Does it matter? It would in an ion pumped TEM, but in an SEM? } 5 All other suppliers do anodize. That is a better reason than you may think! } 6 If we did not, people would buy other supplier's mounts because ours would } look "cheap" } } Now, I have a question: How long have you and some other people been using } anodized mounts and not realised that the nice shiny finish is not a polish. } Have you never seen machine finished Al??? } } If you really want to make a point, place a hundred commercial mounts into the } bottom of the SEM chamber and then repeat that experiment with machine finished } Al mounts. Record vacuua at various points and run the experiment three times. } Report the results. I would be interested to learn if there is any difference. } Those hundred plain Al mounts made in a machine shop would be a trifle } expensive (you could acid strip the anodizing), but cheaper than most } experiments. } } Christmas is coming, so lets be charitable: anodized mounts are a good deal and } they are very suitable for SEM work. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } } On Monday, December 03, 2001 12:14 PM, Frederick Schamber } [SMTP:schamber-at-aspexllc.com] wrote: } } I would certainly hope that SEM stubs are NOT anodized. Not only does } } anodizing } } create an insulating surface, but it also creates a surface with very poor } } vacuum } } properties -- anodized parts will pump down slowly due to the gas trapped in } } the } } porosity. } } } } Fred Schamber } } ASPEX, LLC } } } } Jim at Proscitech wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } I'm certain that every mount sold by ProSciTech is anodized, and by their } } } very } } } appearance, all other such commercially sold mounts I have seen are } } } anodized } } } too. True the anodized film is very thin. } } } Cheers } } } Jim Darley } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } } ABN: 99 724 136 560 www.proscitech.com } } } } } } On Saturday, December 01, 2001 9:07 AM, Darrell Miles } } } [SMTP:milesd-at-US.ibm.com] } } } wrote: } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Anodizing of aluminum can be done in many colors, as } } } } well as a clear coating. I would not expect SEM stubs to be } } } } anodized, as the aluminum oxide coating which forms is an } } } } insulator. This would inhibit the removal of the electron } } } } charge from the sample. The native oxide film is no where } } } } near the thickness of the usual anodized film, and can easily } } } } be broken. The thickness of the anodized film can be } } } } controlled, but it is still an effective insulator. I am fairly } } } } certain that none of the stubs I have ever used were anodized. } } } } } } } } Regards, } } } } Darrell
Surely the most famous misinterpretations of microscopical observations are those recorded in drawings made by preformationists in the 18th Century of the "homunculus" in the head of human sperm. Several of these drawings are commonly reprinted in textbooks as cautionary examples.
For a discussion of a common histological artifact see "Multinucleate Plant Cells" by Burkholder and Mc Veigh (1941) in vol. 68 of the Bulletin of the Torrey Botanical Club p. 395.
You should also check out what the web has to offer on the history of Royal Rife and the Rife Microscope.
Finally, evidence seems to be accumulating that the "nanobacteria" observed by EM in the martian meteorite are artifacts.
Mike Dalbey
Lecturer in Biology University of California, Santa Cruz
Some subscribers to the list did not see this message the first time I posted it. At Nestor's suggestion, I am re-posting it, with apologies to those who did see it the first time!
Tony G-R
-------------
The New England Society for Microscopy (NESM) will be holding it's 35th Annual Fall Symposium at UMASS-Lowell in Lowell, Massachusetts on Friday, Dec. 7, 2001.
The meeting will begin at 12 Noon with Registration in the Mil Conference Center-Wannalancit Building (North Campus).
There will be 3 Sessions: A special student session beginning at 1:05 pm with 4 presentations, followed by a Poster session and coffee break.
Session II (Biological) begins at 2:20 pm and has as it's speaker, Ken Moore (University of Iowa), the National MSA Speaker. His talk will be: "Application of Microscopy Techniques to the Study of Genetic Therapy Research".
Another Poster Session and Afternoon Coffee Break will follow at 3:20 pm.
Session III (Materials Science) will begin at 3:40 pm and have 2 speakers: Elen Humphreys from M.I.T. and Koichi Nishikida from Thermo Spectra-Tech.
The Annual Business Meeting will convene at 5:00 pm and will include the election of NESM officers for 2002.
A Dinner and after-dinner speaker will follow at 6 pm.
Advance registration is due by Friday, November 30th. Registration after November 30th will NOT include dinner. The registration for NESM members is $10.00 and $25.00 for non-members. Dinner is $15.00 extra.
For more detailed information, please contact Mary McCann, NESM Treasurer at mccanns-at-tiac.net or NESM's website: http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
---------- From: Griffiths, Vern Sent: Friday, November 30, 2001 11:34 AM To: 'microscopy-at-sparc5.microscopy.com' Subject: RE: Cleaning SEM stubs
We have a metallographic lab next door. I've cleaned and reused hundreds of stubs. After I've accumulated 50 - 100 stubs, I put them in an ultrasonic cleaner in acetone for five to ten minutes. This gets off most carbon paint and adhering specimen stuff and the like. Then I take the stubs, one at a time, and polish/grind them on a 320 grit silicon carbide paper on a standard metallographic polishing wheel. I generally do the sides and one end, sometimes (rarely) both ends and they look like new. A relatively negligible amount of material is removed so that I have stubs on which this operation has been performed many times. Each stub may require 30 - 60 seconds total so that you may find this time consuming but for us it's worth the effort. I don't know what the black stuff you mention is, but aluminum will corrode in aqueous solutions and the Alconox is probably alkaline enough to aggravate the situation. Vern Griffiths, Montana Tech ---------- From: jmkrupp-at-cats.ucsc.edu {mailto:jmkrupp-at-cats.ucsc.edu} [SMTP:jmkrupp-at-cats.ucsc.edu] {mailto:[SMTP:jmkrupp-at-cats.ucsc.edu]} Sent: Thursday, November 29, 2001 5:35 PM To: Microscopy-at-sparc5.microscopy.com {mailto:Microscopy-at-sparc5.microscopy.com} Subject: Cleaning SEM stubs ------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html}
I heard on the radio that the economy is in a recession, so I figured I had better get busy and try to save money and recycle some supplies. I also had a big box of used SEM stubs and thought I could start there.
The stubs are left over from various projects, some have silver paint, some with double stick tape, etc. I thought I could loosen things up a bit by soaking them in water and/or an Alconox solution. This fit in very well with my plan for recycling since the plan was to let them soak until the recession is over.
Since there was no way to know when things would return to normal, I put some into an ultrasonic cleaner with Fisher Ultrasonic Cleaning solution. Says it is safe for aluminum and other things.
Now I have a problem. The stubs turned pretty ugly. Big black stains all over the place. Looks like some reaction between the Al and the Alconox. Stubs in plain water are not so bad. Ultrasonic cleaner can't remove the black stuff. I can polish off the black stuff, but that wasn't part of the plan. This was supposed to be simple.
I don't think the black stuff affects the functioning of the stub, it just looks ugly and means I have to explain to some users that they are just as good as the few shiny ones left in the drawer (stubs that is, not users).
So tell me oh mighty metal and materials experts, what is the black stuff, why did it appear, and is there any easy way to get rid of it? Or is this some kind of plan to pull the economy back up to speed by making me buy a bunch of shiny new SEM stubs?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu {mailto:jmkrupp-at-cats.ucsc.edu}
John M. Basgen wrote: =============================================================== We have been using carbon grating replicas for calibrating the magnification of our TEM images for many years. We are in the process of revising our imaging and morphometry protocols. I am not happy with the precision and longevity of the carbon grating replicas. Are there more precise and/or stronger standards for TEM magnification calibration? The final magnifications of our images range from 3,000 -20,000 X. ================================================================ Have you seen the MAG*I*CAL™ TEM Calibration Specimen as described on the SPI website, page URL http://www.2spi.com/catalog/standards/magical.html
It seems to get around most of the problems associated with the carbon grating replicas and although nothing is forever, it is quite robust in comparison and should last quite a long time.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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As far as the finish goes, I have machined aluminum myself, and have left a far "shinier", superior finish on the aluminum, compared to what the normal SEM stubs show. I must admit that I have never had any from ProSciTech, but from your statement, they are very similar. It must be more than the finish that makes you spend the money to have them anodized.
True, the freshly machined aluminum forms a native oxide fairly fast, but it is nowhere near the thickness of the oxide formed by anodizing.
I'm not trying to be uncharitable, I am just surprised that someone would go to the expense of anodizing SEM stubs, which also forms an insulating film. I have made my own mounts for specific uses which are not anodized, and they don't look any different from the commercially acquired ones, even after years of use. Of course, the proper alloy needs to be used, or you WILL have vacuum problems.
Food Structure & Functionality Forum Symposium 2002 May 5 to 8, 2002, Palais des Congres de Montreal, Montreal, Quebec, Canada
An international symposium leading Food Structure & Functionality studies through the 21st century
"webaddress at the AOCS site (http://www.aocs.org/member/division/fsff/index.htm)", bulletin board (http://www.aocs.org/ubbcgi/ultimatebb.cgi
held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
The mandate of the Food Structure & Functionality Forum : "To promote global collaboration between Food and Agriculture professionals in Structure and Functionality disciplines by facilitating and providing a forum for exchange of knowledge, expertise and research findings". The symposium has two themes: * new and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Technical Program Schedule (as of December 3rd, 2001)
Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl)
Monday, May 6th-Morning Opening of symposium - Opening remarks
Plenary Speaker and presentation of Division Achievement Award
Dairy Applications Session. Chairs: Mark Auty, Dairy Products Research Centre, TEAGASC, Ireland (mauty-at-moorepark.teagasc.ie ) and Harjinder Singh, Massey University, NZ (H.Singh-at-massey.ac.nz)
Some Observations of a Microscopist, Author, and Reviewer on Structural Studies of Milk Products. M. Kalab, Agriculture and Agri-Food Canada, Canada
Effect of Emulsifiers and Processing Conditions on Microstructure of Milk Fat/Sunflower Oil Blends. S. Martini1, M. Cerdeira1, C. Puppo1, R.W. Hartel2, and M.L. Herrera1,3, 1CIDCA, UNLP, CONICET, Argentina; 1University of Wisconsin-Madison, USA; 3University of Buenos Aires, Argentina
Texturization of Dairy-Based Spreads. Y. Shi, B. Liang, and R.W. Hartel, University of Wisconsin-Madison, USA
Localization of Whey and Casein in Cheeses Using Microscopy and Immunochemistry Techniques. Y. Wang and D. Pechak, Kraft Foods, U.S.A
Manufacturing Yoghurt Structures with a Predicted Consumer Preference. M Langton and A. Astrom, SIK, Sweden
Dynamic Confocal Imaging of Tension and Fracture in Composite Food Materials. D.P Ferdinando1, K.P Plucknett2, and V. Normand3, 1Unilever Research, UK; 2DERA, UK; 3Firmenich SA, Switzerland
TBA - Topic: Dairy powders/caramels. C. Attapattu, University of Wisconsin, USA
Monday, May 6th - Afternoon Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl ) and David Pechak (Dpechak-at-kraft.com)
Protein Polysaccharide Interactions. C.G. De Kruif, NIZO Food Research, Netherlands (keynote speaker)
To Be Announced. B. Campbell, Kraft Foods, USA
Structure in Heat Treated Low_Fat Emulsions. R. Ofstad and V. Hoest, MATFORSK, Norway
Fatty Acid Salts-Induced Gels of Food Proteins: Their Rheological Properties and Structural Changes of the Proteins During Gelation. N. Yuno-Ohta, Nihon University, Japan
Wheat Gluten Proteins. A.S. Tatham, IACR Long Ashton research Station, United Kingdom
Interfacial Composition and Stability of Oil in Water Emulsion formed with Mixtures of Milk Proteins and Polysaccharides. H. Singh and Y. Hemar, Institute of Food, Nutrition and Human Health, Massey University, NZ
Dedicated Poster Session
Division Board Meeting
Tuesday, May 7th - Morning Agricultural Applications of Microscopy and Imaging Session/ joint with Feed Microscopy Division. Topic: Food Contamination contacts: Mark Auty, Dairy Products Research Centre, TEAGASC (mauty-at-moorepark.teagasc.ie ) and Marge McCutcheon, West Virginia Department of Agriculture, USA (Feed Microscopy Division)
Contaminants in Food Processing. D. Kittleson, General Mills Technology East , USA
How to approach contaminant identification. M. Auty, Dairy Products Research Centre, Ireland
Identification of plant material. D.F. Wood, USDA, USA
To Be Announced. J. Makowski, Windsor and Associates, USA
Species identification of Animal Hair by Using Atomic Force Microscopy. C.W. Cruywagen, University of Stellenbosch, South Africa.
Quanitation of Total Fat and Fat Quality in Cheese and Dairy Products Using Membrane Separation Technology. V.C. Gordon, Safety Associates, Inc., USA
Detection and Differentiation of APRALAN, PAYLEAN, PULMOTIL and TYLAN in Animal Feeds using microscopy. P. Klink. Elanco Animal Health, a Division of Eli Lilly and Company, USA.
Additonal speakers to be announced.
Division Luncheon and round table (expert) discussion. Topic to be announced
Tuesday, May 7th - Afternoon Microbiology and Food Session Chairpersons: Judy Arnold and Ida Yates, USDA, ARS, Russell Research Center, USA
Prevention of Bacterial Fouling on Food Equipment Surfaces. J.W. Arnold, USDA, ARS, Russell Research Center, USA
Probiotics and Their Use in Food Animal Production. R. Droleskey, USDA, ARS, SPARC, USA
The Effect of High Pressure Steriliztion on Listeria Inoculated Seafood. K.R.S. Schneider and M.V.W. Wood, University of Florida, USA
Food Microstructure Investigations by Atomic Force Microscopy. J. Thornton, Digital Instruments/Veeco Metrology Group, USA
Controlling Growth of the Toxigenic Fungus, Fusarium Verticillioides. I. Yates and J. Arnold, Russell Research Center, ARS, USDA, USA
Division Members Meeting (immediately following the afternoon session)
Wednesday, May 8th- Morning Ingredients and Food Processing Session- Chairpersons: Diana Kittleson, General Mills Technology East, USA; and Bernhard Tauscher, Federal Research Center for Nutrition, Germany
Non-Invasive Quality Determination of Fruit and Vegetables: Application of a Multi-Wavelength NIR-Diode Laser Array. B. Tauscher, Federal Research Center for Nutrition, Germany
Characterisation of the Application of Novel Oil Structurants. E. Floeter, F. Gandolfo, and W. Hogervorst, Unilever Research Vlaardingen, The Netherlands
High Pressure Processing. E. Ting and E. Raghubeer, Flow International, USA
Microstructure of Rice Starch Isolates. D.F. Wood1, A.M.Ibanez_Carranza2, and C.F. Shoemaker2, 1USDA, ARS, WRRC, USA; 2University of California, USA
To be announced, D.W. Stanley, Department of Food Science, University of Guelph, Canada
To Be Announced. F. Escher, B. Conde-Petit, ETH, Switzerland
To Be Announced. M. Michel, Nestec Ltd., Nestle Research Center, Switzerland
To Be Announced. M. Salmenkallio-Marttila , VTT Biotechnology, Finland
To Be Announced. J. Boye, Agriculture and Agri-Food Canada, Canada.
Wednesday, May 8th - Afternoon New Methods and Techniques for Food Structure and Functionality Analysis SessionChairpersons: Kathy Groves, Leatherhead Food Research Association, England; and Maud Langton, SIK, Sweden
Quantifying Microstructures through Image Analysis. G.M.P. van Kempen1, M. van Ginkel2, C.L. Luengo Hendriks2, L.J. van Vliet2, and S. Singleton3, 1Unilever Research Vlaardingen, Netherlands; 2Delft University of Technology, The Netherlands; 3Unilever Research Colworth House, Great Britain
Cryo-TEM of Biopolymers in Comparision with Other TEM-techniques. M. Langton, A. Altskar, and A.-M Hermansson, SIK, Sweden
Freeze-substitution and low temperature embedding of dairy products for electron microscopy. A.K. Smith and H.D. Goff, Department of Food Science, University of Guelph, Canada
MicroRheology: preliminary results of structural behaviour of foods under deformation. M. Paques, Y. Nicolas, Wageningen Centre for Food Sciences/Unilever Vlaardingen, The Netherlands.
Recent Advances in our Understanding of the Relationship Between Crystallization Behavior, Microstructure and Rheological Properties of Fat Crystal Networks. A. Marangoni, University of Guelph, Canada
Spectroscopic Prediction of Rheological Properties in Grains. F. Meadows and F. Barton USDA, ARS, QARU, USA
Staining Techniques for Detection of Components in Fish Muscle. K. Hanneson Eggen and G. Enersen, Matforsk, Norway
Changes in plant tissue after pulsed electric field treatment. M. Fincan, P. Dejmek, Dept. of Food Engineering, Lund University, Sweden
Closure of Symposium
Posters
Relationships between Microstructure and Rheological Properties of Model Lipid Systems. B. Liang, Y. Shi, and R.W., Hartel Univ. of Wisconsin-Madison, USA
Minor Biomolecules from the Olive Drupe to Olive Oil: The Technology and the Well-being Effects. N. Uccella, CIRASAIA-Mediterranean Agrifood Research Centre, Calabria University, Italy
Formation and Physical Properties of ?-Fat Gel. I: Macroscopic and Microscopic Observations. K. Higaki1, Y. Sasakura1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. Ii: In situ Observation of Gel-Formation Processes. K. Higaki1, Y. Sasakura1, I. Hachiya1, S. Ueno2, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. III: Rheological Properties. K. Higaki1, T. Koyano1, I. Hachiya1, and K. Sato2, 1Meiji Seika Kaisha Ltd., Japan; 2Faculty of Applied Biological Science, Hiroshima University, Japan
Formation and Physical Properties of ?-Fat Gel. IV: Why and How? K. Sato1, K. Higaki2, T. Koyano2, and I. Hachiya2, 1Faculty of Applied Biological Science, Hiroshima University, Japan; 2Meiji Seika Kaisha Ltd., Japan
Exchange in Semi-Solid Triglyceride systems measured by NMR Spectroscopy: Effect of Partial Glycerides on Exchange Rates. P. Smith1, N. Haghshenas1, I. Furo2, and B. Bergenstahl3, 1YKI Institute for Surface Chemistry, Sweden; 2Royal Institute, Sweden; 3Lund University, Sweden.
Effect of Shear Rate on Fat Crystallization Kinetics. P.H. Rousset and V. Mooser, Nestle Research Center, Switzerland.
Utilizing Polarized Light Microscopy to Characterize the Effects of Tween60 on the Physical Properties of a Model Plastic Fat System. J.W. Litwinenko and A.G. Marangoni, University of Guelph, Canada.
AnnFook Yang may have gone to the experts, but do they know their own products?
The material known today as Sodium Thiosulphate (or sodium thiosulfate), with the composition Na2S2O3.5H2O was formerly known as sodium hyposulphite. As it was the principal ingredient in fixers, the conventional abbreviation for the fixing bath was "Hypo". The chemistry hasn't changed - just the name has been forgotten, but not by Kodak, who still manufacture (or at least, did last time we ordered it!) "Hypo-Clear", which is a rapid washing agent.
Kodak's current Rapid Fixer (which isn't listed on their Web site, but is referred to in the literature accompanying their current black-and-white photography products, and which we and others still buy in significant quantities) uses Ammonium Thiosulphate rather than the sodium salt. The same literature also specifies the use of Kodak Fixer, which presumably therefore is still available, and which I imagine is still a sodium thiosulphate (or "Hypo"!) based fixer, but I don't have any on hand, so can't be sure of that.
Interestingly, Ilford's web pages also do not mention their black-and-white fixers - only by going to the product specification pages can one find mention of the Ilford line of B&W chemicals - as asides in the processing recommendations. Is this some sort of conspiracy to render the market obsolete by denying users any information about the products, thus ensuring that the demand will wither away?
For our film - SO163 in our case - we use the exact sequence that the Kodak support person recommended, as we have for the past 25 years. For paper, on the rare occasions when we make photographic enlargements, we add the step of soaking in Hypo-Clear.
Tony Garratt-Reed
} I have never used Orbit bath nor Hypo. In fact the former is totally new } to me. I have heard of Hypo in the '60s. I was curious and went to the expert. } I phoned Kodak Scientific Support (800-225-5352) to find out whether } developing the 4489 film in D19 and fixing in Kodak Rapid Fixer is } adequate. The answer is "Yes" } What about Hypo? He asked, "Who makes it?" } He assured me that nothing else is needed after Rapid Fix. Just wash the } negs. in water will do. } } } } } } AnnFook Yang } EM Unit, } Eastern Cereal and Oilseed Research Centre, } Room 2091, Bldg. 20, } Central Experimental Farm, } Ottawa, Ontario } Canada K1A 0C6 } } Tel: 1-613-759-1638 } Fax: 1-613-759-1701 } } e-mail: yanga-at-em.agr.ca } } } } } "Paula Sicurello" {patpxs-at-gwumc.edu} 11/30/01 03:25PM } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Catherine, a microtome devoted to polymers should have a cryo chamber since a lot of polymers are easier to section at sub-zero (C) temperatures. Definitely recommend a 35 deg knife. Other than that, just use common strategy for microtomy of biological samples (with some patience for trial/error temp setting). You can contact me off-line for our instrument info. Good luck,
Alice.
Alice Dohnalkova Battelle, PNNL Richland, WA (509)372-0692
-----Original Message----- } From: "Catherine.Derr-at-gastechnology.org"-at-sparc5.microscopy.com [mailto:"Catherine.Derr-at-gastechnology.org"-at-sparc5.microscopy.com] Sent: Monday, December 03, 2001 6:20 AM To: Microscopy-at-sparc5.microscopy.com
I looking at purchasing a microtome for sectioning polymers, primarily PE. Although I am familiar with microtomes used in biological studies, I have no experience in sectioning polymers. Has anyone sectioned polymers with repeated success? What features are desired for sectioning polymers? Is there a specific knife angle that works well?
KD Derr SEM/Polymer Technician Gas Technology Institute 1700 S Mount Prospect Rd Des Plaines, IL 60018 (847) 768-0505 (phone) (847) 768-0569 (fax) kd.derr-at-gastechnology.org
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Catherine Derr wrote: ====================================================== I looking at purchasing a microtome for sectioning polymers, primarily PE. Although I am familiar with microtomes used in biological studies, I have no experience in sectioning polymers. Has anyone sectioned polymers with repeated success? What features are desired for sectioning polymers? Is there a specific knife angle that works well? ====================================================== We have been sectioning polyethylene for over thirty years in our laboratory . If you are talking about thin sectioning polyethylene, unpigmented, then in many ways, the sectioning is not going to be greatly different from your biological samples.
There are several things that are going to be very important for you:
1] PE sectioning can not be done at room temperature, only cryo.
2] polyethylene is very soft, and the key is to do the sectioning with minimum distortion. That means diamond knives, and the smaller the angle the better. We usually are using a 35° knife angle.
3] polyethylene is either injection molded (into parts) or melt extruded (into film). The results you get from sectioning along the injection or extrusion direction might not necessarily be the same as when you section transverse to that section. So you want to give some thought as to which view would give you the most useful information considering your objectives and then it would be most important to always section all samples at the same angle relative to the machine direction. Otherwise, differences in section angle could be "interpreted" as differences between samples but such would be false interpretations.
4] pure unfilled polyethylene are normally pretty unremarkable by thin section TEM because there is nothing really to "see" or to give any contrast . Unless you were trying something exotic with the sections, such as negative staining of the lamellar or spherulytic structures, your main concern would be in the sectioning of the inorganic additive particles which in general would be much harder than the polyethylene. The presence of these additives would impart small damge points on the knife edge which would then show up as annoying striations in the final section (and image). Therefore you don't want to use your good (and expensive) life science knives for such sectioning, but a good materials science knife (which can be obtained through SPI or the other firms offering diamond knives.
5] We have found that when comparing microtomes from the two major ultramicrotome manufacturers, one can get comparable results using either system. Just remember that for this kind of work, a good cryo system is needed.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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I would like to do 3D reconstructions out of my .avi or .tif series, is it possible to do so with the Confocal Assistant ? If not, what easy-to-handle software is available?
Thank you
Albert Cardona Genetics Department University of Barcelona Av. Diagonal, 645 08028 Barcelona, Spain
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As a follow up to your posting on anodised componenets in vacuum - decorative anodising is often wiped with a light oil or polish, after dyeing, to give that nice shiny look.
Now that really isn't good for the vacuum!!!!
Regards, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
For crying into a big bucket. Under the onslaught I have no doubt that normal heavy duty anodized surface (like a handrail) are unsuitable for SEM mounts. } Most aluminium anodization that I have seen creates a dark } THICK oxide film (tens to hundreds of nm thick) and is insulating. (Nestor).
But I never suggested that the mounts were heavily anodized; on the contrary I stated that the coating on our mounts is very thin. Furthermore, I stated that the appearance of our mounts is identical to those from other suppliers. Recently, because of a lost shipment we had to buy some American-made mounts and I'm sure that no customer would have noted.
I am not an expert on anodizing either, but when many years ago I sent samples to our manufacturer, I was immediately advised - "they are anodized". So we had ours "anodized", afterall the samples had come from Chuck!
I find it amazing that, seemingly same listers believed that I would supply dull, thick anodized mounts which don't conduct and outgas catastrophically. My previous two communications, made it clear that that just wasn't so.
The issues may be: Is our supplier electropolishing, but calls that anodizing? Or how would in practice a very thin layer of anodizing differ from the naturally occurring oxide film? Or what is the end result of electro-polishing? Could this in fact add a thin anodized layer? Is a very thin anodized film shiny or dull?
I would have liked answers to those questions, which could have resolved the differences. At this point I know that the mounts are not just machined Al; they had a treatment and according to the manufacturer they are "anodized". There is of course the possibility that our manufacturer never applied any process. But why would he? The charge for "anodizing" is paltry. Thank you for your vigorous discussion. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, December 04, 2001 4:01 AM, Frederick Schamber [SMTP:schamber-at-aspexllc.com] wrote: } Jim, } I will be the first to admit that I am not an expert on either SEM stubs or } anodizing, but I have had very negative experience with the latter in vacuum } systems. I once designed some relatively small black-anodized parts into a } SEM and spent considerable time trying to find out why we had such bad } "outgassing" -- finally, someone wiser than me sanded off the anodizing and } the vacuum was fine. So I have, in effect, done the experiment you describe } -- ordinary machined aluminum parts are vastly superior to anodized aluminum } parts in terms of their vacuum properties in my experience (and I did some } followup experiments in a well-controlled apparatus designed specifically to } measure vacuum properties of materials and confirmed that a little bit of } anodizing is a very bad thing). It is not a question of the mere "roughness" } of the surface. Apparently the anodizing process creates very tiny } interstices which hold the dye, and this is what causes the problem -- or so } I have assumed. I suppose it could be the dye itself -- I never tried the } experiment on "clear anodized" parts. } When I look at catalog advertisements for aluminum SEM stubs, I always see } reference to various degress of polishing, and don't recall ever seeing } reference to anodizing. } From an electrical standpoint, I also really wonder how anodizing could work. } I know that anodizing must be avoided on electrical cabinetry because one } cannot establish reliable contacts (a different surface treatment process is } used on aluminum instead). I'm sceptical about the "thin coating" concept } since it is my understanding that it is precisely the insulating layer which } protects the surface from oxidation. Any amount of insulation on a SEM stub } would be a big problem. (Of course, the same thing can be said about the thin } oxide layer that naturally forms when aluminum is exposed to air.) But it is } my understanding that anodizing is a process which modifies the surface, not } a simple "coating" which can be "thin". } My negative opinions about anodizing for vacuum systems aren't just mine -- in } fact, I thought they were "common knowledge". For example, I refer you to the } very informative vacuum information site maintained by Roy Schmaus at the } University of Alberta } ~http://nyquist.ee.ualberta.ca/~schmaus/vacf/index.html } {http://nyquist.ee.ualberta.ca/schmaus/vacf/index.html} } Under Basics/an Introduction to Materials for use in Vacuum we find the } following statement: "Aluminum that will be exposed to vacuum should never be } anodized due to serious outgassing problems." Under "References" on the same } page he notes: "I recently had a call from someone who has used Anodized } Aluminum as an insulator down into the ultra high vacuum region. Admittedly } the pieces used were very small and initial outgassing was a problem but low } costs were a major advantage." [emphasis added] } So given my negative experience and the congruent remarks of others who work } in this field, I have long assumed that anodizing was something to avoid at } all costs in vacuum systems -- and I've assumed that applied to SEM stubs. Am } I wrong? } How do polished stubs stay so shiny? I've wondered the same. However, anodized } parts I have seen always have a kind of "matte" appearance, rather than a } high gloss in any case. } So, though it is entirely possible that I will learn something from this } discussion that I did not know previously (it would hardly be the first } time), my limited grasp of the subject still convinces me that SEM stubs } should NOT be anodized. But how do they retain their surface finish and } conductivity? Your response suggests that you manufacture SEM stubs and call } out an anodized surface -- did I read this correctly? Or are you purchasing } stubs which you assume are anodized? I really would be interested if someone } who is in the business of manufacturing these things could comment directly } to this -- without disclosing proprietary information of course. } "Charity" in view of the season notwithstanding, I consider this a matter of } considerable practical importance. If I am in error about this, I want to } know it, since a non-insulating, non-outgassing, but still protective } anodizing surface would be a very good thing. If I am correct, then others } reading this exchange should know that anodizing is something that should be } scrupulously avoided in vacuum systems (SEM and otherwise). In either case, } getting to the bottom of this will presumably be a good thing for everyone. } I'm prepared to learn from new facts, if someone can put them on the table. } Fred Schamber } ASPEX, LLC } } Jim at Proscitech wrote: } You are hoping in vain, commercial mounts are anodized, because: } 1 The layer is too thin to insulate even at low voltages. SEM have high volts } } (even specimen currents are many and not fractions of volts) and low currents. } } No problem. } 2 Otherwise the mounts would oxidise and I believe that too is not the best } conductor. } 3 Al too is porous, if you really care use polished stainless. } 4 Does it matter? It would in an ion pumped TEM, but in an SEM? } 5 All other suppliers do anodize. That is a better reason than you may think! } } 6 If we did not, people would buy other supplier's mounts because ours would } look "cheap" } Now, I have a question: How long have you and some other people been using } anodized mounts and not realised that the nice shiny finish is not a polish. } Have you never seen machine finished Al??? } If you really want to make a point, place a hundred commercial mounts into the } } bottom of the SEM chamber and then repeat that experiment with machine } finished } Al mounts. Record vacuua at various points and run the experiment three times. } } Report the results. I would be interested to learn if there is any difference. } } Those hundred plain Al mounts made in a machine shop would be a trifle } expensive (you could acid strip the anodizing), but cheaper than most } experiments. } Christmas is coming, so lets be charitable: anodized mounts are a good deal } and } they are very suitable for SEM work. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } ABN: 99 724 136 560 www.proscitech.com } On Monday, December 03, 2001 12:14 PM, Frederick Schamber } [SMTP:schamber-at-aspexllc.com] wrote: } } I would certainly hope that SEM stubs are NOT anodized. Not only does } } anodizing } } create an insulating surface, but it also creates a surface with very poor } } vacuum } } properties -- anodized parts will pump down slowly due to the gas trapped in } } } } the } } porosity. } } } } Fred Schamber } } ASPEX, LLC } } } } Jim at Proscitech wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } } } } } } -----------------------------------------------------------------------. } } } } } } I'm certain that every mount sold by ProSciTech is anodized, and by their } } } } } } very } } } appearance, all other such commercially sold mounts I have seen are } } } anodized } } } too. True the anodized film is very thin. } } } Cheers } } } Jim Darley } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } } ABN: 99 724 136 560 www.proscitech.com } } } } } } On Saturday, December 01, 2001 9:07 AM, Darrell Miles } } } [SMTP:milesd-at-US.ibm.com] } } } wrote: } } } } ------------------------------------------------------------------------ } } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } } } } } On-Line Help } } } } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } Anodizing of aluminum can be done in many colors, as } } } } well as a clear coating. I would not expect SEM stubs to be } } } } anodized, as the aluminum oxide coating which forms is an } } } } insulator. This would inhibit the removal of the electron } } } } charge from the sample. The native oxide film is no where } } } } near the thickness of the usual anodized film, and can easily } } } } be broken. The thickness of the anodized film can be } } } } controlled, but it is still an effective insulator. I am fairly } } } } certain that none of the stubs I have ever used were anodized. } } } } } } } } Regards, } } } } Darrell
The consensus on my question was HUH? Most of the EM world has used a hypo clearing agent after the fixer step to reduce the washing time. Though I did get a reply stating that is was used originally to reduce the time in fixer and it was supposed to make for better archival properties.
I will be returning to the tried and true method as soon as it gets here.
Thanks to all who replied.
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Just to clear up a couple of statements someone made. There is no such thing as a "clear" anodic film. And dies are not used to give anodic films colour ..The colour comes from metals in the anodic layer. Thickness of the layer is also a factor.
Paul D. Nolan Electron Optics
Alcan International Limited Kingston Research and Development Centre P.O.Box 8400, 945 Princess Street Kingston, Ontario K7L 5L9
The native oxide thickness of aluminum metal, depending upon processing, is typically on the order of 30 to 50 angstroms. You can measure this thickness very easily by making a simple XPS measurement of the Al2p core level.
I'm looking for a double sided adhesive to stick samples (aluminum) onto glass slides. I want to be able to remove them with relative ease and replace them for storage. Conductivity is not an issue At present i am using those little double sided sticky press down adhesives used for SEM stubs ..they are a little to sticky for my liking.
Any suggestions?
Paul D. Nolan Electron Optics
Alcan International Limited Kingston Research and Development Centre P.O.Box 8400, 945 Princess Street Kingston, Ontario K7L 5L9
} With any instrument purchase the best thing to do is go to the } manufacturers, or better yet have them come to you, and see how the } instrument works on your samples. I have extensive experience sectioning } PE and I have always used Reichert-Jung, now Lieca, ultramicrotomes. That } does not mean others would not do the job. The main thing is to get a } system that works well when cryo-ultramicrotoming. As stated in another } response you may well have to cryotome your samples and if you are buying } for polymer work you definitely want to get a system that will work at } both room temp and cryo conditions. How to section PE can differ } depending on the type of PE, high density or low density. If you have } HDPE then you can often pre-stain with Chlorosulfonic Acid and room temp } microtome. Otherwise you may have to stain with RuO4 before or after } cryomicrotoming. There are many good references on these } techniques. Good luck on your purchase.
} I looking at purchasing a microtome for sectioning polymers, primarily } PE. Although I am familiar with microtomes used in biological studies, I } have } no experience in sectioning polymers. Has anyone sectioned polymers with } repeated success? What features are desired for sectioning polymers? Is } there a specific knife angle that works well? } } KD Derr } SEM/Polymer Technician } Gas Technology Institute } 1700 S Mount Prospect Rd } Des Plaines, IL 60018 } (847) 768-0505 (phone) } (847) 768-0569 (fax) } kd.derr-at-gastechnology.org
Stephen McCartney Research Associate 2108 Hahn Hall Materials Institute Virginia Tech Blacksburg, VA 24061-0344 USA
Being a Plant Pathologist, the best I can think of is the story of Pierce's disease of grapes, which was of unknown etiology for a long time. Forgive me for not seeking details from the literature.
At one time it was proposed to be caused by a virus. Extensive light microscope work missed the true cause- a bacterium. After elucidation of the pathogen, I am told that upon review of work previously done, the bacterium WAS there to be seen, but was missed.
Upon a similar vein, MLO's in plants were not recognized in TEM work until a human or animal pathologist saw micrographs of MLO's in plant tissue, and was readily able to say what they were.
Anecdotal, but food for thought.
Lesson? I better keep my eyes and my mind open.
Maureen Petersen Dept. Plant Pathology University of Florida
Dear Listers, Given that my curiosity is up per the plating/no plating of the stubs, my posting is geared more towards the actual reusing of stubs. For the past eight years I have been subjecting a set of 50 or so stubs to a sand blasting treatment. You may question the feasibility of a sandblaster BUT what is important is the media used to blast with. glass beads in the range of 170-325 mesh are used to effectively remove residue, glue, tape, and any material that resides on the stub. This particular media results in no residue and no tolerance change, although a pure water treatment and ultrasonic wash are utilized. I have had no trouble at all even after this amount of time.
Robert Fowler Quality Assurance Technician (Failure Analysis) TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.tdk.com
I embed the hair in epoxy after the usual fixation and dehydration steps. Then section with a glass knife, mount on a slide, and stain with toluidene blue. Joyce Craig Chicago State University
You are right about an anodized aluminum finish. It is an excellent insulator, not at all the type of finish you want on an SEM specimen mount, as conductivity is critical. I think it may also create vacuum problems.
The mounts (stubs) we manufacture are machined on very high speed turning equipment from high grade aluminum bar stock. After machining, we put them through a rigorous part cleaning process using special cleaning solutions. Typical chemical based solutions tend to leave a film on the mounts that both insulates and degrades vacuum. Event our turning equipment uses a special, non-petro based cutting lubricant to avoid the potential for any contamination. Each mount is then carefully inspected and wiped down to ensure that they are completely clean and dry, then stored in air tight packing material.
Because our machines operate at such high speeds, our process eliminates the need for polishing. However, mounts that are made on more conventional lathes or machines may require a final polishing to achieve the same results.
I hope this helps.
Best regards, Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: zaluzec-at-sparc5.microscopy.com [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Monday, December 03, 2001 9:32 AM To: Microscopy-at-sparc5.microscopy.com
As I recall from my metallurgy
To "Anodize" is to subject a metal to an electrolytic action as the anode of a cell in order to coat/deposit a protective or decorative film.
Most aluminium anodization that I have seen creates a dark THICK oxide film (tens to hundreds of nm thick) and is insulating.
I've never seen this on an aluminium SEM stub. Certainly many I've used are electropolished and some (the ones we make here in our machine shop ) are simply milled and mechanically polished. All will at a minimum have a native oxide. All are conductive, and yes (Jim) I've seen polished (electrochemical, mechanical) as well as all ranges of "machine" finished products. These are NOT anodized, they are all simply oxidized by leaving them sit in air. The native oxide which forms is a few tens of angstroms thick and is stable, by definition this is not anodization. I have never heard of anyone calling this process "anodization" should that be what you are referring to.
So... now I'm curious and I would request that the various suppliers who like reply to this list to fill us all in on what you do.
1.) How many of you anodize your stubs (ie. coat), vs electropolish, vs mechanically polish. 2.) If you intentionally anodize, (ie. deposit/coat) your stub with an electrodeposited layer how thick is your film, and what do you anodize it with. 3.) It might also help to define the "anodization" process (if you do this) so that we are all understand what each of you mean in the context of how you make the stubs we purchase.
we inadvertantly ordered several thousand 4x5 inch glassine envelopes (for EM negatives). As we are not using this size and I would hate to waste them, anybody could have them for free. I am at the University of Chicago. Please contact me offline.
Paul Is there a distinction between an anodic film and an anodised film??
Organic dyes are in fact widely used to colour anodized aluminium, though I am prepared to concede that I am not up to date with techniques for anodizing, and may have missed recent innovations involving metal-doping of the oxide film. The aluminium oxide film formed on anodized aluminium at first has a frosted appearance, which is presumably caused by light scattering from multiple voids and micro-rough surfaces. The film is rendered more translucent by various post-anodizing treatments, following dyeing if desired. But once this treatment is carried out, the film is clear - i.e. you can see right through to the base metal, aluminium oxide being one of nature's clear, light-transmitting materials. Most people have experienced lightfastness problems in dyed anodized aluminium products, a syndrome which I am not aware occurs in metal-doped alumina such as ruby. I have seen the anodizing process being carried out, including dyeing with organic dyes, without any other metal than aluminium in sight. Was I dreaming this?
Chris
} Just to clear up a couple of statements someone made. } There is no such thing as a "clear" anodic film. } And dies are not used to give anodic films colour ..The colour comes from } metals in the anodic layer. Thickness of the layer is also a factor. } } Paul D. Nolan } Electron Optics } } Alcan International Limited } Kingston Research and Development Centre } P.O.Box 8400, 945 Princess Street } Kingston, Ontario K7L 5L9 } } Tel: (613) 541-2066 } Fax: (613) 541-2134 } paul.nolan-at-alcan.com } }
Robert, I also bead-blast stubs. Works very well as long as you're not looking for a smooth surface, then you have to grind and polish afterwards, but the bead blasting DOES clean that top surface in a hurry. On Al I usually only blast at 35psi to keep the erosion down.
I have several pieces of Al that have a lot 1/8" holes drilled in them. Load'em up with stubs and blast away! It's fast, it's fun and reasonably ecologically sound. Follow with a BRIEF ultrasonic bath in JOY, rinse with tap, DI or distilled water, depending on how particular you are, and dry. Alconox and some of the other heavy-duty lab cleaners have a pH that is far too high for Al, although they work great on glass.
Ken Converse owner Quality Images third party SEM service Delta, PA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } Given that my curiosity is up per the plating/no plating of the stubs, my } posting is geared more towards the actual reusing of stubs. For the past } eight years I have been subjecting a set of 50 or so stubs to a sand } blasting treatment. You may question the feasibility of a sandblaster BUT } what is important is the media used to blast with. glass beads in the range } of 170-325 mesh are used to effectively remove residue, glue, tape, and any } material that resides on the stub. This particular media results in no } residue and no tolerance change, although a pure water treatment and } ultrasonic wash are utilized. I have had no trouble at all even after this } amount of time. } } Robert Fowler } Quality Assurance Technician (Failure Analysis) } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.tdk.com } } } }
Michael R. Nesta wrote: =============================================================== You are right about an anodized aluminum finish. It is an excellent insulator, not at all the type of finish you want on an SEM specimen mount, as conductivity is critical. I think it may also create vacuum problems.
The mounts (stubs) we manufacture are machined on very high speed turning equipment from high grade aluminum bar stock. After machining, we put them through a rigorous part cleaning process using special cleaning solutions. Typical chemical based solutions tend to leave a film on the mounts that both insulates and degrades vacuum. Event our turning equipment uses a special, non-petro based cutting lubricant to avoid the potential for any contamination. Each mount is then carefully inspected and wiped down to ensure that they are completely clean and dry, then stored in air tight packing material.
Because our machines operate at such high speeds, our process eliminates the need for polishing. However, mounts that are made on more conventional lathes or machines may require a final polishing to achieve the same results
"Paul.Nolan-at-alcan.com"-at-sparc5.microscopy.com wrote: } Just to clear up a couple of statements someone made. } There is no such thing as a "clear" anodic film. } And dies are not used to give anodic films colour ..The colour comes from } metals in the anodic layer. Thickness of the layer is also a factor. } Paul D. Nolan
The anodizing process for aluminium indeed does require a dye. There are plenty of companies selling the dyes etc.. http://www.caswellplating.com/frames.asp?bottom=/anodizedye.htm http://www.plating-supplies.com/Anodizing.html http://www.davistl.com/services/anodizing_plating.htm
Some pages actually showing the process: http://www.focuser.com/atm/anodize/anodize.html http://hem.passagen.se/ballista/anod_eng.html
And one does get the "clear" anodization by using same process, but not using dye.
There are as well other methods for getting, for example titanium, colourfull surface films. These are used in jewelry etc. and some do refer to these as anoziding (anodic polarization). However, the aluminium anodizing does as common term refer to the dye process described above.
Not sure if this one fits the bill but it is a similar one to the grape disease story by Maureen Petersen.
The one I mean is the discovery/description of Helicobacter pylori in human gastric biopsies in the early 80's (when it was given the name Campylobacter pylori). It had been there all of the time but had been disregarded as 'stuff'.
I seem to recall a similar story regarding the finding of Actinomyces in cervical smear material.
We tend to see what we expect might possibly be there and consciously or unconsciously ignore the rest as 'noise' - perhaps a natural action but also potentially dangerous.
I have not looked up the original references either but a search on www.google.com will provide much information.
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
We usually tend to avoid issues between suppliers which are controversial, but based on our long experience in the business in this area, we shall. Ladd, being the oldest EM supplier and first company to provide commercail SEM mounts, has never anodized our mounts. We continue to machine polish them as we have done thru the years. Based on our long history we feel that anodizing would be ineffective at best and perhaps just increase cost unnecessarily.
John Arnott
Disclainer: Ladd Research sells EM supplies, including SEM mounts. --
**** Please Note Our New Address, Fax and Phone Numbers ****
LADD RESEARCH 83 Holly Court Williston, VT 05495 USA
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Quality Since 1955
zaluzec-at-sparc5.microscopy.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } As I recall from my metallurgy } } To "Anodize" is to subject a metal to an electrolytic action as the } anode of a cell in order to coat/deposit a protective or decorative film. } } Most aluminium anodization that I have seen creates a dark } THICK oxide film (tens to hundreds of nm thick) and is insulating. } } I've never seen this on an aluminium SEM stub. Certainly many I've used are } electropolished and some (the ones we make here in our machine shop ) } are simply milled and mechanically polished. All will at a minimum } have a native oxide. } All are conductive, and yes (Jim) I've seen polished } (electrochemical, mechanical) as } well as all ranges of "machine" finished products. These are NOT anodized, they } are all simply oxidized by leaving them sit in air. The native oxide } which forms } is a few tens of angstroms thick and is stable, by definition this is not } anodization. I have never heard of anyone calling this process "anodization" } should that be what you are referring to. } } So... now I'm curious and I would request that the various suppliers } who like reply } to this list to fill us all in on what you do. } } 1.) How many of you anodize your stubs (ie. coat), vs electropolish, } vs mechanically polish. } 2.) If you intentionally anodize, (ie. deposit/coat) your stub with } an electrodeposited layer } how thick is your film, and what do you anodize it with. } 3.) It might also help to define the "anodization" process (if you do } this) so that we are all } understand what each of you mean in the context of how you make the } stubs we purchase. } } Nestor } Your Friendly Neighborhood SysOp.
--
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LADD RESEARCH 83 Holly Court Williston, VT 05495 USA
On-line Catalog: http://www.laddresearch.com
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Listers, I know this topic has come up previously but many of the responses have gone directly to the inquirer rather than to the list. I am looking for information about video rate CCD camera for TEM below $50,000 in cost. The intended use for the camera is teaching and assisting investigators in viewing samples in a service lab situation. It is not meant for capture of high quality images but capture of lower quality images "for the record" would be helpful. I am already in touch with Gatan concerning such cameras so would like to hear about other options. I would appreciate hearing from users regarding their recommendations and experiences but do not mind hearing directly from venders. Thank you, Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
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Ken, If a smoother finish is in order then I also use a more aggressive media (60 - 120 mesh). This media gives a much smoother "glossy" finish but I personally have never had a reason to have the surface that smooth.
Robert Fowler Quality Assurance Technician (Failure Analysis) TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.tdk.com
Ken Converse {qualityimages-at-n To: "robert.fowler-at-tdktca.com" etrax.net} -at-sparc5.microscopy.com, "MSA, listserver" {Microscopy-at-sparc5.microscopy.com} 12/04/2001 05:34 cc: PM Subject: Re: Aluminum Stubs
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Robert, I also bead-blast stubs. Works very well as long as you're not looking for a smooth surface, then you have to grind and polish afterwards, but the bead blasting DOES clean that top surface in a hurry. On Al I usually only blast at 35psi to keep the erosion down.
I have several pieces of Al that have a lot 1/8" holes drilled in them. Load'em up with stubs and blast away! It's fast, it's fun and reasonably ecologically sound. Follow with a BRIEF ultrasonic bath in JOY, rinse with tap, DI or distilled water, depending on how particular you are, and dry. Alconox and some of the other heavy-duty lab cleaners have a pH that is far too high for Al, although they work great on glass.
Ken Converse owner Quality Images third party SEM service Delta, PA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } Given that my curiosity is up per the plating/no plating of the stubs, my } posting is geared more towards the actual reusing of stubs. For the past } eight years I have been subjecting a set of 50 or so stubs to a sand } blasting treatment. You may question the feasibility of a sandblaster BUT } what is important is the media used to blast with. glass beads in the range } of 170-325 mesh are used to effectively remove residue, glue, tape, and any } material that resides on the stub. This particular media results in no } residue and no tolerance change, although a pure water treatment and } ultrasonic wash are utilized. I have had no trouble at all even after this } amount of time. } } Robert Fowler } Quality Assurance Technician (Failure Analysis) } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.tdk.com } } } }
One of my customers needs to remove AFM cantilevers from SEM stubs after examination without breaking them. Standard SEM adhesives are too sticky for him too.
We cut a square from the adhesive section of a Post-It note, and used double-stick tape to attach it to the stub with the Post-It adhesive facing up. It is sticky enough to hold his small samples in place - if your samples are not too large or heavy, it might be worth a try.
The Department of Energy has issued a solicitation for Small Business Innovative Research (SBIR) proposals related to electron beam instrumentation. This is a great opportunity for small businesses to apply for research funding to help develop their creative ideas for instrumentation and methods that can help advance the field. This is the second year the solicitation includes electron beam instrumentation and it is hoped that a strong set of proposals will be received.
The web site detailing the solicitation is: http://sbir.er.doe.gov/sbir/Solicitations/FY%202002/BES.htm#T9 (This will take you directly to the portion dealing with electron beam instrumentation.)
Electron beam instrumentation is included in technical topic 9, "NEUTRON AND ELECTRON BEAM INSTRUMENTATION." Part b covers Electron Beam Microcharacterization Facilities.
In short, grant applications are sought:
-to develop stages and holders with new capabilities for in situ experiments in the transmission electron microscope.
-to develop electron sources for scanning transmission electron microscopy with brightness on the order 109 Amp/cm2/steradian or higher.
-for systems for automated data collection and processing.
-for improved x-ray and electron detectors.
The closing date is Jan 15, 2002 but please be sure to check the web site to verify this!!
Please distribute this information to your colleagues that may have an interest in this area and encourage them to think creatively. If there are any questions, please contact the SBIR program office or you may contact me off-line.
Best regards, Dean
--------------------------- Dean J. Miller Materials Science Division Argonne National Laboratory Argonne, IL 60439
We use cleaved edge specimens of III-V multilayers. We can measure layer thicknesses in-house to a good accuracy (~0.1%) using high resolution X-ray diffraction, whereas the mag-i-cal is (I believe) only guaranteed to +/- 2%. Measurement errors from TEM negatives puts the error in calibration up to about 0.5%. The thin area on the samples is miniscule and they are very robust. A drawback is that you need to be able to tilt the sample to 45 degrees. As discussed some time ago on this listserver, you need to be careful to eliminate lens hysteresis effects if you want to make your measurements accurate.
Cheers,
Richard Beanland Marconi Caswell Ltd. Towcester, Northants, NN12 8EQ UK
e-mail richard.beanland-at-marconi.com
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: 03 December 2001 18:53 To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
John M. Basgen wrote: =============================================================== We have been using carbon grating replicas for calibrating the magnification of our TEM images for many years. We are in the process of revising our imaging and morphometry protocols. I am not happy with the precision and longevity of the carbon grating replicas. Are there more precise and/or stronger standards for TEM magnification calibration? The final magnifications of our images range from 3,000 -20,000 X. ================================================================ Have you seen the MAG*I*CAL(tm) TEM Calibration Specimen as described on the SPI website, page URL http://www.2spi.com/catalog/standards/magical.html
It seems to get around most of the problems associated with the carbon grating replicas and although nothing is forever, it is quite robust in comparison and should last quite a long time.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Anodizing. Seems like everyone knows what it is, until they compare notes!
The furor drove me to crack a wonderful older textbook, Thin Film Processes, edited by John L. Vossen and Werner Kern, Academic Press 1978.
} From my reading the reason for the confusion is clear: there really are a lot of ways to anodize aluminum, some of which result in a clear coating, some colored, some porous, some hard. The most common and least expensive method is the sulfuric acid process with follow-up dying described so excellently on the web pages Kristian Ukkonen refers us to.
For anyone who is interested, here are some excerpts on various ways of anodizing aluminum, from the chapter by Frederick Lowenheim on "Deposition of Inorganic Films from Solution".
"Although H2SO4 is the most common anodizing electrolyte, many others are in use. For special effects or attainment of special properties, chromic acid anodic coatings are opaque, limited to about 10 um in thickness...
"Phosphoric acid anodizing has been used as a basis for plating on Al, though it has been largely superseded by the zincate and other processes.
"Oxalic acid coatings are yellow, and somewhat harder than conventional H2SO4 coatings. Mixtures of oxalic and sulfuric acid produce hard coatings, competing with low-temperature H2SO4 processes.
"Sulfonated organic acids, combined with H2SO4, produce so-called 'integrally colored' coatings on specific alloys. Various shades are used in architectural applications...
"Boric acid electrolytes, often with additions of borax, produce thin barrier oxide coatings used for electrical capacitors.
"Many anodic coatings on Al can be colored, either with organic dyes or mineral pigments. Postanodic treatments, especially that known as sealing, are important in determining the final properties of the coating..."
Libby Shaw
} From: Kristian Ukkonen {kristian.ukkonen-at-iki.fi} } } } "Paul.Nolan-at-alcan.com"-at-sparc5.microscopy.com wrote: } } Just to clear up a couple of statements someone made. } } There is no such thing as a "clear" anodic film. } } And dies are not used to give anodic films colour ..The colour comes from } } metals in the anodic layer. Thickness of the layer is also a factor. } } Paul D. Nolan } } The anodizing process for aluminium indeed does require a dye. } There are plenty of companies selling the dyes etc.. } http://www.caswellplating.com/frames.asp?bottom=/anodizedye.htm } http://www.plating-supplies.com/Anodizing.html } http://www.davistl.com/services/anodizing_plating.htm } } Some pages actually showing the process: } http://www.focuser.com/atm/anodize/anodize.html } http://hem.passagen.se/ballista/anod_eng.html } } And one does get the "clear" anodization by using same } process, but not using dye. } } There are as well other methods for getting, for example } titanium, colourfull surface films. These are used in } jewelry etc. and some do refer to these as anoziding } (anodic polarization). However, the aluminium anodizing } does as common term refer to the dye process described } above. } } Kristian Ukkonen. } *************************************************************** Elisabeth L. Shaw, Facility Coordinator Surface and Spectroscopy Labs Analytical Shared Experimental Facilities MIT Center for Materials Science and Engineering
Address: MIT Room 13-4149 Tel: 617-253-5045 77 Massachusetts Ave. Email: elshaw-at-mit.edu Cambridge, MA 02139 Fax: 617-258-6478 http://web.mit.edu/cmse/www/ ***************************************************************
This query is directed specifically at users of the Hitachi S4700 FESEM, but may also apply more widely to users of FESEM's.
We get a curious result (to me) when flashing the tip. We were advised to flash until the emission current reading read about 20uA on the "le" meter. The flash intensity adjustment behind the scope was initially set to accomplish this in one or two flashes when flashing at an intensity of "2" in the Setup-Column menu.
That is what happened for quite some time, however for a while now flashing has resulted in values between 7-12 on the "le" meter. The initial flash briefly degrades the vacuum at the gun as expected as contamination is driven off, but subsequent flashes do not, also as expected. However, subsequent flashes usually LOWER the "le" meter value, rather than raise it toward our goal of 20. This is the really puzzling part to me. Originally, the value increased with each flash, and if an excessive number of flashes were required, we would adjust the intensity pot in the rear of the scope.
We normally run the scope at an emission of 10uA, with kV's of 1.0-10.0. All vacuum readings are normal, and the scope performs beautifully in terms of resolution and brightness.
My questions are:
1) Why is the "le" value decreasing with multiple flashes?
2) How dangerous are multiple flashes to the tip?
3) Does the condition described above indicate any problem with the tip and/or scope?
4) Is flashing actually accomplishing anything if the vacuum gauge shows no contamination being driven off?
5) Is it safe to keep turning up the intensity pot on the rear of the scope? (Something tells me no.)
6) Do I simply not understand what's going on? (Something tells me yes.)
Thanks in advance!
Puzzled in Missouri, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
FYI: ctfexplorer has been updated and it is still free for all. New features/improvements: - extended microscope list - "Euro-bug" removed (problem with Windows locales which use a comma instead of a point for a decimal separator).
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98/NT4/2000.
Please give it a try. Please DO direct your suggestions and comments to sidorov-at-yahoo.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer (always use this link. it will direct you to the right location). Direct link: http://www.maxsidorov.com/ctfexplorer
Enjoy, __________________________________ Max Sidorov, Ph.D. sidorov-at-yahoo.com
----------Additional Info---------- ctfExplorer is a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfexplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
Features - Calculates 1-Dimensional CTF - Calculates 2-Dimensional CTF - Calculates Defocus Map - Calculates point-to-point resolution, Lichte defocus and info limit - Shows the effects of 2-Fold and 3-Fold astigmatism - Allows to change Defocus, 2-Fold and 3-Fold astigmatism in real time - Shows what happens to 1D CTF in different directions when there's astigmatism - Displays the damping envelopes - Allows to select a microscope from a list of microscopes - Allows to create a custom microscope - Allows to change HT, Cs, Cc, Energy Spread, Convergence for any microscope - 2 modes of operation: CTF Explorer (to see everything) and CTF Plotter - Compares 2 microscopes or 2 settings for 1 microscope - Saves 1D CTF, 2D CTF and "Defocus Map" as bitmaps or metafiles - Exports 1D plots to tab-delimited text format
What you are looking for sounds like our product numbers 60550, Double Coated Aluminum tape. Please contatc me or check our website for more details.
John Arnott
Disclaimer: Ladd Reserch sells EM supplies. This product may be available from other vendors. --
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Paul.Nolan-at-alcan.com-at-sparc5.microscopy.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hello } } I'm looking for a double sided adhesive to stick samples (aluminum) onto } glass slides. } I want to be able to remove them with relative ease and replace them for } storage. } Conductivity is not an issue } At present i am using those little double sided sticky press down adhesives } used for SEM stubs ..they are a little to sticky for my liking. } } Any suggestions? } } Paul D. Nolan } Electron Optics } } Alcan International Limited } Kingston Research and Development Centre } P.O.Box 8400, 945 Princess Street } Kingston, Ontario K7L 5L9 } } Tel: (613) 541-2066 } Fax: (613) 541-2134 } paul.nolan-at-alcan.com
Hello, I have another vacuum problem with our Electronscan E3 ESEM after swapping out a failing rotary pump. The vacuum was working fine and then after being in standby for a couple days, we went into wet mode and had a vacuum failure with the message: "Error cannot Determine state" on the console. Resetting gives the same message. Power cycling the console gives an "E2 can't pump column error", followed by the cannot determine state message.
Powering everything off and restarting, it seems the diffusion pumps won't start up now. Everything was working fine, but now I think there is a problem with the vacuum control unit or a sensor. Is there a way of resetting the VCU, or some other troubleshooting I can do? Thanks for any advice. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Thanks for the comment, Mike. Just a bad wording from my side. MAx
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov] Sent: Wednesday, December 05, 2001 10:56 AM Cc: sidorov-at-yahoo.com
Sorry about the subject name, it is a bit misleading. In using a Leica SP2 confocal, I have noticed that it is possible to see the cover slip and slide. Why/how is this possible? The setup is essentially this: a coverslip is placed on a slide, which is then placed on the stage (there is nothing between them, except of course for miscellaneous small dust particles and air). The slide is then scanned with light from a HeNe laser (633nm). When the light detection (wavelengths directed to the PMT's) is set to include 633nm (i.e. 610-650nm), the signal is of course very bright. I am assuming that this is due to reflection. What I am confused about is that if the light detection doesn't include 633nm (sampling from 660-710nm), there is still signal present, and it is in the same pattern as is seen when looking at the reflected light. It is important to note that this pattern is connected to the coverslip and slide somehow, as moving the stage will cause a corresponding change in the image produced by the system. I realize that I may be missing something fairly obvious, but any help would be greatly appreciated.
Thanks, Russell
-- Confocal Imaging Facility Technician Tufts Medical School 136 Harrison Ave. Boston, MA 02111 Tel. (617) 636-3795
Leslie has a great idea. I've used it myself for wafer pieces. Another thing that I've used is the double-sided carbon tape, but have reduced the surface adhesion by touching it repeatedly with my fingers or something else that is strong enough to be pried off without damage. This makes the tape tacky, but without the death grip is has when it's new.
Leslie Eibest wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Paul; } } One of my customers needs to remove AFM cantilevers from SEM } stubs after examination without breaking them. Standard SEM } adhesives are too sticky for him too. } } We cut a square from the adhesive section of a Post-It note, } and used double-stick tape to attach it to the stub with the Post-It } adhesive facing up. It is sticky enough to hold his small samples in } place - if your samples are not too large or heavy, it might be worth } a try. } } Leslie Eibest } SEM lab
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) SC Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I have commented on advantages of using the MAG-I-CAL sample in the past and will not do so again here.
I want to comment on Richard's response below. Before I do so, I want to say that I am an overly enthusiastic supporter and believer in the Small Angle Cleavage Technique (SACT) or MicroCleave(TM) Technique and that South Bay Technology is the only company that makes it and I don't work for them. (I hope that this helps keep any feathers from being ruffled.)
Richard, SACT is a marvelous way of preparing III-V samples very quickly and it is an excellent way of calibrating layer thickness values in the TEM when used in conjunction with high resolution X-ray diffraction. I have a PDF file on the SBT web site (https://www.southbaytech.com) that illustrates how this is done. Search for the Microcleave Kit (model number 520) and then look at the application note number 61. The advantage that the SACT method has is that you can get all the thickness that you need from very thin to very thick. By setting up the appropriate imaging conditions, thickness of the layers can be measured very accurately. Similar to what you have said in your response, you can use them to calibrate the TEM. This is what John McCaffrey did when he created the Mag-i-cal sample. BTW, application note #226 is a poster of the technique that John and I put together and is also available on the SBT web site.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-marconi.com] Sent: Wednesday, December 05, 2001 9:39 AM To: Microscopy-at-sparc5.microscopy.com
We use cleaved edge specimens of III-V multilayers. We can measure layer thicknesses in-house to a good accuracy (~0.1%) using high resolution X-ray diffraction, whereas the mag-i-cal is (I believe) only guaranteed to +/- 2%. Measurement errors from TEM negatives puts the error in calibration up to about 0.5%. The thin area on the samples is miniscule and they are very robust. A drawback is that you need to be able to tilt the sample to 45 degrees. As discussed some time ago on this listserver, you need to be careful to eliminate lens hysteresis effects if you want to make your measurements accurate.
Cheers,
Richard Beanland Marconi Caswell Ltd. Towcester, Northants, NN12 8EQ UK
e-mail richard.beanland-at-marconi.com
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: 03 December 2001 18:53 To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
John M. Basgen wrote: =============================================================== We have been using carbon grating replicas for calibrating the magnification of our TEM images for many years. We are in the process of revising our imaging and morphometry protocols. I am not happy with the precision and longevity of the carbon grating replicas. Are there more precise and/or stronger standards for TEM magnification calibration? The final magnifications of our images range from 3,000 -20,000 X. ================================================================ Have you seen the MAG*I*CAL(tm) TEM Calibration Specimen as described on the SPI website, page URL http://www.2spi.com/catalog/standards/magical.html
It seems to get around most of the problems associated with the carbon grating replicas and although nothing is forever, it is quite robust in comparison and should last quite a long time.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (microscope-at-tin.it) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, December 5, 2001 at 09:25:51 ---------------------------------------------------------------------------
Email: microscope-at-tin.it Name: Massimo
Organization: None
Education: Graduate College
Location: Torino; Italy
Question: 05 December 2001
Let introduce myself. I am an Italian amateur naturalist. I have a light microscope and I have arranged an iris diaphragm on the collector lens of illuminator system to performing an illumination according to Koehler method. I do not understand well the last procedure step:
{ { Step 4. Now lift the eyepiece out of the body tube and look down the tube at the back lens of the fully-lighted objective. (This is best accomplished by the use of a pinhole eyepiece; an eyepiece with a tiny hole) while looking down the microscope tube, slowly open and close the substage condenser aperture iris diaphragm. It will be seen that closing the aperture iris diaphragm "cuts into" the periphery of the back lens of the objective. For excellent illumination and contrast, approximately º-1/3 of the back lens should be occluded, thus leaving 2/3-3/4 of the back lens illuminated. Then replace the eyepiece in the tube. This step too must be repeated each time a different objective is turned into place on the nosepiece.} }
So just a few questions:
how much would the hole diameter be of the pinhole eyepiece? And in which way would I have to use it? Or what would I expect by seeing throught the hole?
There are some photos or web location where I could see the back lens to become occluded by closing or opening the aperture iris diaphragm? And to understand the meaning of this operation.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (robert.s.mattingly-at-grc.nasa.gov) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, December 5, 2001 at 14:00:05 ---------------------------------------------------------------------------
Email: robert.s.mattingly-at-grc.nasa.gov Name: Bob Mattingly
Organization: NASA Glenn Research Center
Education: Undergraduate College
Location: Cleveland, Ohio, USA
Question: I have heard of an adapter for mounting a Nikon 995 Digital Camera to an Olympus SZH-ILLD Macroscope. I cannot find any information on the manufacturer of an adapter to meet my application. Are you aware of any suppliers? Thanks in advance for your help with this issue.
Our Leica Orthoplan 2 needs to be decontaminated. Fungi have grown on the front surface mirrors in the internal prism of the main stand.
In the past, we had to return the stand to Germany for repair but are wondering if anyone knows of a qualified Leica repair center in the US. Barring this, we would purchase a second Orthoplan 2 main stand if one were available.
Thank you.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
It is a while since our E3 departed, so my memory of the symptoms is a bit rusty. However, a problem we had from time to time was the "U" bend in the vibration isolation block in the pumping lines filling with oil, and thereby preventing any pumping from taking place, once the column has been pumped to a modest vacuum. This would cause the vacuum pumps to fail to start as you described.
The remedy was to disconnect the rubber hoses, pour out the oil, then reassemble and start up.
I do recall seeing the error messages you describe, but whether they were associated with this particular fault I can't now remember.
Tony Garratt-Reed.
At 10:58 AM 12/5/2001 -0800, Gordon Vrololjak wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
They make nice adapters for the 990 to most any scope.
gary g.
At 03:29 PM 12/5/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all who responded to this query. There is much more to this than meets the eye. Rather typical in this field it seems.
Finding a sharp edge seems easy enough. Not so. Sharp is sharp but really sharp is hard to find. At 200KX, sharp is not so sharp. I used some xxxx maker's apertures only to find that they were not so sharply defined. I tried two others and found that they too were not sharp. I finally found one of my Amray 70u Au coated apertures to be sharp enough to use. I had to tilt it to make a truly sharp edge. The trick is to increase contrast, decrease brightness and slow scan the image at 200KX. Then, do the line scan. I go the following results:
With my Amray FESEM, higher CL# means smaller spot size. So the spot size numbers seem to jive.
This is no small feat to measure!! The key is to do the slow scan first and then do the line scan. Otherwise, there is too much mechanical noise and human interference (coughing) which corrupts the results.
I'm going to do CL15 and CL20 tomorrow and see what I get.
I intuitively see the rationale for how this works. It becomes more obvious as I work with it. The Gaussian aspects are surprising yet rather logical. A 3D plot of a spot would surely look like this. -- good stuff.
A recent example image is at http://photoweb.net/spot-23.tif
Thanks.
gary g.
At 03:53 PM 11/26/2001, you wrote: } The SEM spot size can be measured by doing a line scan across a sharp edge. } For more details see the SEM lab workbook by Lyman et al., 1990. } } Krassimir Bozhilov. } } } } } } Hi all: } } } } Is there any direct way to quantitatively determine } } and measure the actual spot size of a SEM beam? } } } } Given various KV, WD, final aperture sizes, condenser lens } } settings, etc., is there a way to truly find out what } } the real spot size is on the specimen? } } } } Any ideas? } } } } gary g. } } } }
Hi, Alan and Listers- } } I am interested in gaining insight into the role of microscopic artifacts } in the history of biology.
Although I 'm sure there are examples of misinterpretation of artifacts that have been perpetuated for many years, I can offer here a story of the flip side. This is about what appeared to be a well-known artifact that turned out to be, in fact, a surprising real structure. It's also an embarrassing personal story!
An undergraduate student was working with some colleagues who were studying the behavior and neurophysiology of the escape response of several species of planktonic copepods (small crustaceans). They had reams of data and were beginning to characterize the responses of the animals to different kinds of stimuli. Each species had it's own characteristic response, but a general picture was beginning to form. There appeared to be two different classes of response though, one type particularly puzzling. The undergrad spent some time in the EM facility with me, using SEM to describe the sensory setae. She expressed an interest in trying TEM and so we went through fixation, embedding, sectioning, etc. I had worked on the tiny beasts for close to 10 years and had found some interesting structures that I worked on from time to time. Copepods were difficult to fix, some species more so than others. So I had concentrated on the "pretty" ones and temporarily given up on the "ugly" ones - the ones with all those myelin body artifacts that I just couldn't seem to avoid. Finally it was time to try to work on them again, so I gave this one particularly difficult species to the undergrad to try to fix and section. She came to me with micrographs, wondering what in the world were those squiggles she saw. I patiently explained that myelin bodies were a fixation artifact, produced when lipids became mobile during fixation, then reformed in these "onion bodies" . I sent her back to work to try again, and she came up with the same results. "What if they aren't an artifact?" she kept asking. I explained that invertebrates do not have such membranous structures; they were artifactual. I brought out the books and papers about myelin body artifacts. I brought out the books and papers that said invertebrates do not have membranous wrappings around their axons. She was unconvinced. In a hurry one day when she was really bugging me I dug through my files and found the decade-old folder to show her I'd seen the same thing - an artifact. "But there's an axon in the middle of each one", she protested, not knowing that was "impossible". I glanced at them impatiently and saw that she was right, but she was running off to class.
Later that day we looked closely and decided that, indeed, the images were a real mess and difficult to interpret, but there was a faint possibility that these weren't classic myelin body artifacts. Whatever it was was certainly reproducible, and these forms appeared every time in the antennal nerve. So we emailed the PIs of the project and told them we might have myelinated axons in the difficult-to-interpret species. It was April1, April Fools' Day, the undergrad's name was April, and they thought it was a joke. After all, the dogma is that invertebrates don't have myelin! Check any biology book.
But by the next day they realized that myelin (OK, "myelin-like structures" for you purists) would instantly and completely explain 10 years' worth of puzzling data.
It took awhile to prove to ourselves (mainly me, since I'd been repeating the dogma for many years) that these structures were real, orderly, and extensive wrappings of membrane around the axons. It took ultrarapid cryofixation and cryosubstitution to really show that the artifactual bodies that had plagued a number of people working on these critters were in truth these surprising, dogma-busting structures.
We made the April the first author on the Nature paper, which came out in April 1999.
I don't take anything for granted anymore! The lesson is: keep an open mind.
For more on the copepod story- Copepod neuroecology http://www.pbrc.hawaii.edu/~petra/copepod.html
Aloha, Tina
http://www.pbrc.hawaii.edu/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hi There, I'm interested in mesuring specimen thickness in TEM. In my case these two techniques (covergent beam technique and the spacing Moire franges) can not be applied. I'll apreciate any further suggestion. Thanks in advance M. Jouiad
-------------------------------------------------------- DR. Mustapha JOUIAD
.. does anbody have an idea (... or even better plans) how to build a more or less "homebrewn" probecurrentmeter (... FEGSEM -} pA resolution needed) ... a comercial instrument is probably to expensive??? (} = 10TDM,- = 5000€,- ???) .. i just had to learn that this is a very useful tool!!!
Hi, Mustapha, you could try EELS for that; the method can give you a very precise value for local thickness.
Corneliu Sarbu, PhD MTMDept. Belgium
- - - - - - - - - - - - - - Original Message - - - - - - - - - - - - - - ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi There, I'm interested in mesuring specimen thickness in TEM. In my case these two techniques (covergent beam technique and the spacing Moire franges) can not be applied. I'll apreciate any further suggestion. Thanks in advance M. Jouiad
-------------------------------------------------------- DR. Mustapha JOUIAD
Thanks to everyone who responded to my request for information about flatbed scanners! There appears to be a number of very good instruments out there. Thanks for your suggestions!
Ken --
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323
I would not think that specimen current is independent of spot size. In fact, I go to great lengths to adjust KV, WD and spot size to obtain just the right specimen current to image upper metal on microchips underneath 10000A of SiN. I lay down a Schottky barrier for the particular type of passivation and adjust the SEM parameters for best picture. Too high of specimen current produces too many SE and thus charging and bad images. The optimum is just enough SE for detail and plenty of BSE for topography.
I can measure specimen current using my Fluke DMM. If the value is not too low, the Fluke will read it (nA). I also use it to measure gun brightness as part of routine calibration.
I'll try measuring the currents today and see what gives.
gary g.
At 06:00 AM 12/6/2001, you wrote: } Hi } } I am very interested with this topic, and we have done such mesures years } ago, as we buid our lab-made LEED optics, with low energy guns, but with } spot sizes of microns... I have for our Jeol 840 a table buid with } such mesures by an japanese ingenior of Jeol Europe, which gives the spot } size versus WD, Energy, and current. So for exemple, with 10 keV, 10E-11A } and 8 mm WD, it should be 110 angstrom. It's a w type gun. This table is } of course not valuable for an other SEM, and I'll have to built it for our } FE-SEM... } } So, a question : do you have a mesure of the probe current (faraday cup, } or as sample current on carbon tape), corresponding to the condenser } settings you have used ? In case our FE-SEM (Jeol 6700F), Jeol claims that } the spot size is independant of the probe current, and I want to verify } it. And comparaison with an other SEM is interesting too. } } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess } 67037 Strasbourg CEDEX } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
I would like to add a comment to John's discussion on calibration samples. I used both the diffraction gratings and the latex spheres prior to John's development of the Mag-i-cal sample. I found that over the overlapping magnification range that both were suitable, they did not agree. Further, I did see changes in the latex spheres at long exposures --I believe it was when I used a lower energy microscope.
I had to do periodic QS9000 (automotive industry equivalent to ISO-9000) calibration procedures for the microscope. With the Mag-i-cal sample, all of the microscope calibrations can be done with one sample very quickly. It is the only way to go.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: John McCaffrey [mailto:john.mccaffrey-at-nrc.ca] Sent: Thursday, December 06, 2001 1:07 PM To: Microscopy listserver
You can also determine specimen thickness with electron energy loss spectroscopy (EELS) by applying the log-ratio technique to the low loss spectra (see Egerton, Electron energy loss spectroscopy in the electron microscope, New York: Plenum, 1996). This technique yields a result in units of the mean free path for inelastic scattering.
Mustapha Jouiad wrote:
} Hi There, } I'm interested in mesuring specimen thickness in TEM. In my case these two } techniques (covergent beam technique and the spacing Moire franges) can not be } applied. I'll apreciate any further suggestion. } Thanks in advance } M. Jouiad } }
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
I have completed many measurements of spot size and specimen current. Tabular data can be found at http://photoweb.net/SEMLog.pdf
This SEM is a Schottky field emission scope. One of the routine periodic checks is to measure the gun brightness. The spec is 12.5nA. This gun is doing 13.81nA and is just a bit higher than the +-1.5nA limits. The method of correcting this is to adjust the extractor voltage by 100 Volt increments and see how the brightness changes after about an hour. Then, once stable, that is the new extractor voltage. Extractor current can vary greatly whereas the gun brightness is quite steady.
Gun brightness is measured with a 4-digit Fluke 79-III DMM connected to the stage alarm BNC jack. The beam is directed into a Faraday cup located on the stage, CL is opened up to -35 and mag is set to 150KX. The voltage reading on the DMM divided by 1E7 (10 Meg Ohms) is the current at the Faraday cup. I used this same method for measuring specimen current with the target aperture for measuring spot size.
The numbers are somewhat believable. However, at low CL values, the specimen current became negative. So I suspect that this method does not work for really low values of specimen currents.
Can anyone comment on the sanity level of the spot size values I have derived? Are these reasonable for a FESEM at 10KV, 4mmWD, and 100u final aperture?
tnx, gary g.
At 06:00 AM 12/6/2001, you wrote: } Hi } } I am very interested with this topic, and we have done such mesures years } ago, as we buid our lab-made LEED optics, with low energy guns, but with } spot sizes of microns... I have for our Jeol 840 a table buid with } such mesures by an japanese ingenior of Jeol Europe, which gives the spot } size versus WD, Energy, and current. So for exemple, with 10 keV, 10E-11A } and 8 mm WD, it should be 110 angstrom. It's a w type gun. This table is } of course not valuable for an other SEM, and I'll have to built it for our } FE-SEM... } } So, a question : do you have a mesure of the probe current (faraday cup, } or as sample current on carbon tape), corresponding to the condenser } settings you have used ? In case our FE-SEM (Jeol 6700F), Jeol claims that } the spot size is independant of the probe current, and I want to verify } it. And comparaison with an other SEM is interesting too. } } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess } 67037 Strasbourg CEDEX } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
I have an old Olympus IMT-2 inverted scope. It has sliders for inserting filters into the light path for fluorescence imaging. Right now I can only image two fluorophores limited by my slider slots. Does anyone know of some way I can image three dyes, i.e. a filter cube or expanded slider system, specifically for the IMT-2?
John, I'd just like to add a couple of points - I'd like to make it clear that I do think that the the mag-i-cal specimen is probably the best commercially available calibration sample. I didn't mean to be disparaging! It's much better than diffraction grating replicas or other commercially available calibration specimens - it beats diffraction grating replicas in that you can use the crystallography to eliminate specimen tilting and is also more stable. Also, both this and the SACT technique have the advantage that you don't need large tilts and could be used in a single tilt holder.
The extent to which you calibrate your microscope depends on what you use it for - you can get a very accurate relative calibration by taking pictures of the same region at different magnifications. You are only then limited by how accurately you can measure features on the negative (0.05 mm in 5cm is readily achievable with the right specimen and a lupe, which gives you 1% relative accuracy). Once you get down to layers below 100 atomic layers thick then of course it becomes impossible to get better than 1% accuracy with HRTEM. Lack of contrast between the different layers can make things even worse at larger thicknesses for some samples.
The 0.1% layer thickness accuracy I quoted was for X-ray diffraction, and for the right sample XRD does beat TEM every time in terms of accuracy. However, most device structures can't be measured using superlattice fringes in XRD and that is where TEM comes in. Of course we're only measuring thickness because it is a measure of device performance; really thin layers, below 100 atomic planes thick, are usually only used as quantum well structures, delta-doping layers or etch stops, the effectiveness of which can usually be assessed by means more relevant to the device performance than layer thickness (e.g. PL, sheet carrier density, overetching). (I am unaware of any applications which need high accuracy from thin layers apart from microelectronics(?))
I do XRD with a 1mm spot on the sample I use for calibration and make a cleaved edge specimen from a region as close as possible to that position. With a 1% thickness variation over a 3" wafer I'm happy that this is truly representative. With lattice matched III-V layers you can grow as thick as you like, and you get better contrast in dark field 002 images. The downside is that the samples are less stable chemically and physically than Si/SiGe which doesn't lend itself to commercial supply (I certainly wouldn't like to put 100 InP cross sections in the post with a bet that more than 50% would arrive unscathed).
Richard Beanland Marconi Caswell Ltd., Caswell, Towcester, Northants NN12 8EQ UK
e-mail richard.beanland-at-marconi.com
-----Original Message----- } From: John McCaffrey [mailto:john.mccaffrey-at-nrc.ca] Sent: 06 December 2001 18:07 To: Microscopy listserver
Hello John,
While I noted that you weren't too happy with the replica samples, these calibration samples (particularly the cross grating replica samples) are probably still the best choice for the calibration ranges in which you are working. A common cross-grating replica (see, for example, http://www.2spi.com/catalog/standards/othertem10.html) has a line density of 2160 lines/mm, which would give you a line spacing on your negatives of 1.39 mm -at- 3000X and 9.26 mm -at- 20,000X. This length would allow you to measure tens of spacings at all of your magnification ranges, and cut your uncertainty down to reasonable levels. While the edge definition of the grating replicas is not the greatest, and while you may have to replace the sample every year or two, these calibration samples are still among the most simple to use, as well as being reliable and reproducible. They are also inexpensive. Other options include latex spheres, which are in the size range that would be useful for your required magnification ranges. The down-side of these samples is that the spheres have a stated average value, so measurements of many spheres are required to lower the uncertainty to the desired level. Also, there is some anecdotal evidence that these spheres change size when exposed to higher electron doses. If your requirement included the entire set of magnification ranges of your TEM, the MAG*I*CAL sample is still the best bet. (Disclaimer: I "invented" the sample, but will attempt to stay objective about it's merits!). Since this sample and all other samples based on crystal lattice spacings rely on fundamental constants of nature, they are the most accurate and precise samples currently available. However, in spite of this glowing pedigree, crystal-based calibration samples still cannot give measurements with better than ~1% uncertainty. The response to your enquiry from Richard Beanland claiming 0.1% accuracy for cleaved III-V multilayers is a bit misleading (not to mention that he disparagingly compared this supposed superior uncertainty to my beloved MAG*I*CAL!). His layer thickness confidence is based on X-ray diffraction (XRD) measurements. XRD gives very accurate layer thickness measurements, but these measurements are averaged over enormous surface areas in TEM terms - the diameter of the x-ray beam, which can be significant fractions of a square mm. This is a very useful measurement for a layer thickness, but this does not translate directly into uncertainty in a TEM. When a semiconductor multiplayer is viewed in cross section, there are a series of contributors to the layer measurement uncertainty: The state-of-the-art TEM's have resolutions of approximately 0.2 nm. The (002) lattice spacings in most semiconductors (the atomic layers parallel to the surface in most wafers) are a little below 0.3 nm. This means that any measurement of a thicker semiconductor layer in a TEM is going to have a "TEM uncertainty" of approximately one atomic layer at each interface. Producing a semiconductor crystal interface that is perfectly, atomically abrupt (even over the relatively small volume of material used in high resolution TEM) is not trivial, and rarely claimed. Any additional atoms from layer A mixed into layer B, or vice versa, will tend to blur the interface when viewed in cross-sectional TEM. This "epitaxial layer" uncertainty is also approximately one atomic layer at each interface. Highly competent crystal growers can minimize, but not eliminate this interfacial intermixing, allowing the combination of the "TEM uncertainty" and the "epitaxial uncertainty" into one atomic layer at each interface. Semiconductor multilayers can only be grown up to a certain critical thickness, at which point the differences in lattice parameter between the adjoining layers becomes so great that the crystal 'relaxes' with the generation of misfit dislocations at the interface. These dislocations need to be avoided in a calibration sample, but the contrasting layer needs to be grown as thick as possible to minimize the influence (i.e., the percentage) of the two points above. Over an entire semiconductor wafer, there will be systematic thickness variations; i.e., layers may be slightly thicker in the middle of the wafer than at the edge. This is a fairly small, but still significant variation - typically less than 1%. Therefore, it becomes important to know where on the wafer your piece of material comes from. So, why can't the "God-traceable" crystal-based TEM calibration samples give measurements with better than ~1% uncertainty? From the combination of uncertainties given above! The thinnest SiGe marker layers in the MAG*I*CAL sample are approximately 10.0 nm thick, and considering their two interfaces to be uncertain in the worse case to approximately one atomic spacing each, this interfacial contribution to uncertainty is about 0.6%. Coupled with the {1% uncertainty across the entire MAG*I*CAL wafer gives a total uncertainty of ~2%, with a safety margin built in. Richard Beanland did correctly imply the important point that, if one wishes to internally calibrate one's own sample against a known lattice spacing on that same sample, the uncertainties will indeed be less. However, 1% uncertainty is a better estimate of the 'best' uncertainty available in a TEM calibration sample, even for a sample based on a fundamental constant of nature. Cheers John
John P. McCaffrey, Ph.D. Institute for National Measurement Standards National Research Council of Canada M-35 Montreal Road Labs Ottawa, Ontario K1A 0R6 CANADA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We use cleaved edge specimens of III-V multilayers. We can measure layer } thicknesses in-house to a good accuracy (~0.1%) using high resolution X-ray } diffraction, whereas the mag-i-cal is (I believe) only guaranteed to +/- 2%. } Measurement errors from TEM negatives puts the error in calibration up to } about 0.5%. The thin area on the samples is miniscule and they are very } robust. A drawback is that you need to be able to tilt the sample to 45 } degrees. As discussed some time ago on this listserver, you need to be } careful to eliminate lens hysteresis effects if you want to make your } measurements accurate. } } Cheers, } } Richard Beanland } Marconi Caswell Ltd. } Towcester, } Northants, } NN12 8EQ } UK } } e-mail richard.beanland-at-marconi.com } } -----Original Message----- } } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] } Sent: 03 December 2001 18:53 } To: MICROSCOPY BB } Subject: TEM Calibration } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } John M. Basgen wrote: } =============================================================== } We have been using carbon grating replicas for calibrating the magnification } of our TEM images for many years. We are in the process of revising our } imaging and morphometry protocols. I am not happy with the precision and } longevity of the carbon grating replicas. Are there more precise and/or } stronger standards for TEM magnification calibration? The final } magnifications of our images range from 3,000 -20,000 X. } ================================================================ } Have you seen the MAG*I*CAL(tm) TEM Calibration Specimen as described on the } SPI website, page URL } http://www.2spi.com/catalog/standards/magical.html } } It seems to get around most of the problems associated with the carbon } grating replicas and although nothing is forever, it is quite robust in } comparison and should last quite a long time. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ==================================================
Hi All, I am attempting to make very accurate measurements of optical coating thicknesses ( {1%), sampling regions a couple of microns or less in size. This isn't possible with SEM or TEM due to interfacial mixing and/or lack of contrast between the layers. (No matter how well calibrated the microscope may be!) One tool which may do the job (and be more relevant to device performance) is a microspectrophotometer such as a 'Nanospec' - or possibly a spectroscopic ellipsometer with very small spot size. Is anyone aware of such a tool I could try in the UK (I am unable to raise the suppliers of Nanospec tools). Alternatively is there another (preferably optical) technique I could use which could also give relevant data?
Mnay thanks for any ideas you may have,
Richard Beanland, Marconi Caswell Ltd., Caswell, Towcester, Northants NN12 8EQ UK
- KE development www.kedev.com range approx 1 pA - 2 µA price approx 1200 €
- Keithley www.keithley.com range approx 0.02 pA - 20 mA price approx 2000 € (model 6485)
sincerely,
Jean Louis HEITZ
CRITT Matériaux-LNE Est 19 rue de Saint Junien, BP 23 F - 67305 SCHILTIGHEIM CEDEX tel. +33(0) 388 191 510 fax +33(0) 388 191 514 jl.heitz-at-critt.fr www.critt.fr
Hello, I have been dealing with SEM/EDS for only 1.5 years and I have a lot to learn, but recently I have been asked a question: What is the EDS depth of penetration on my unit? I work with Amray 1645 and it was upgraded by PGT for imaging and EDS. Could anybody answer the question How deep does the x-ray beam penetrates through a sample and, if it is varies, how to find out actual depth?
hi all, i am doing collagen extractions from corneas and negative staing them to get the lenghts. my problem is that i am having trouble getting the collagen to stick to the formvar. any suggestions. i am on a deadline to work out the details of this project. John
I have an older SEM which over a period of several months seems to repeatedly develop a small droplet of oil on the EDS detector collimator.
It is a diffusion pumped system. The mechanical backing pump is regularly serviced and gets oil changes.
I am inclined to think it is time to clean the diffusion pump and change the oil there. Perhaps over time backstreaming from the mechanical pump has occurred, causing contamination of the diffusion pump oil with mechanical pump oil. The latter, which has higher vapor pressure, is evaporating out of the diffusion pump into the chamber and condensing on the coldest point there (the detector).
Any opinions on whether this is reasonable or not? Any other possibilities anyone can think of? I'd hate to change the diffusion pump oil, which is going to be a big pain, if this is not going to do the trick.
By the way, I'm already planning to install a foreline trap (there currently isn't one).
Thanks,
Wharton
********************************************************** Wharton Sinkler UOP LLC Des Plaines, IL
If the pattern is the same, the signal is probably still reflected light. This signal can be very large compared to fluorescence and it is just blasting through (or around) the filters/prism.
Jim Pawley
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
----- Forwarded by Robert Fowler/TCU/TDK-US on 12/07/2001 09:54 AM -----
"Monson, Frederick C." To: "'robert.fowler-at-tdktca.com'" {fmonson-at-wcupa {robert.fowler-at-tdktca.com} .edu} cc: Subject: RE: Aluminum Stubs - Long Version 12/07/2001 09:48 AM
Thanks for pointing the direction. The only sand blasting I have any recollection of was many years ago when you could take your spark plugs to Pep Boys and use a machine they had for public use. Like the fluoroscope in Buster Brown(?), they too have disappeared.
Thanks again Bob, I appreciate your time and consideration,
Regards,
Fred
} ---------- } From: robert.fowler-at-tdktca.com } Sent: Friday, December 7, 2001 9:03 AM } To: Monson, Frederick C. } Subject: RE: Aluminum Stubs - Long Version } } } Hello again Mr Monson, } In the past you have taken the time to help me with a problem on my SEM } so } I will take the time to fill you in on the details of my procedures. Due } to } liability reasons I have to say that these are my own procedures and I } cannot endorse anyone using these (reference only). All materials required } are readily available from McMASTER-CARR WWW.MCMASTER.COM } } P/N } Price } 1- Self Contained Vacuum 9808T44 450.00 } 2- HEPA Filter 9808T47 89.00 } 3- Blasting Cabinet 33005K71 250.00 } 4-Media (170-325 mesh) 3386K75 37.00 } 5-Media (60-120 mesh) 3386K72 35.00 } } Stubs are placed in an appropriate holder. This will vary slightly with } the } style of stubs used. A piece of 12 inch square of aluminum with 1/8" or } 1/4" holes is fine. This is placed in the blasting cabinet. Vacuum turned } on. Air supplied is usually run at 35 PSI. If residue is thick (most of } the } time some tapes such as double sided carbon tape can be mostly removed } before blasting) then a higher (50 PSI) air pressure is required. Parts } are } blasted until visibly clean. 170-325 mesh media will produce a matte } finish. 60-120 mesh grit will produce a glossier finish and removes } residues faster. After blasting parts are removed and cleaned in } ultrasonic } wash using DI water. Distilled water is fine. This wash is repeated three } times. The following are clips from correspondence about cleaning stubs. } } Robert, } I also bead-blast stubs. Works very well as long as you're not looking } for a smooth surface, then you have to grind and polish afterwards, but } the bead blasting DOES clean that top surface in a hurry. On Al I } usually only blast at 35psi to keep the erosion down. } } I have several pieces of Al that have a lot 1/8" holes drilled in them. } Load'em up with stubs and blast away! It's fast, it's fun and } reasonably ecologically sound. Follow with a BRIEF ultrasonic bath in } JOY, rinse with tap, DI or distilled water, depending on how particular } you are, and dry. Alconox and some of the other heavy-duty lab cleaners } have a pH that is far too high for Al, although they work great on glass. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Hi Robert, } } You are quite correct, this is undoubtedly the "best" way to clean sample } mounts for reuse. However, we have had several concerns, and that is why } we } have not recommended it ourselves when people ask: } } a) In a large central facility, some of the remains of samples could } contain hazardous materials which would then end up contaminating the } media } and in the end, while you have clean sEM mounts, there is created a large } volume of hazardous waste, and } } b) How would you really know that the "cleaned" mounts were really } "clean" } and in a virgin pristine state? In other words, what would be the "test"? } } I would be curiously as to how you approach those two issues. We } ourselves } have to think about liability issues, and that is why we have not } aggressive } promoted this as a means for people to clean and reuse their "used" SEM } mounts. WE even considered offering a recycling service, and doing mount } cleaning here, but I did not know how to protect the health and safety of } my } people when handing the waste mounts from other laboratories. } } Chuck } } PS: Please remember that we are nearly 100% paperless and we would ask } that any reply to this message be by way of the "reply" feature on your } software, so that the entire string of correspondence can come back to us } and all be in one place. } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } } } Hello Mr Garber } In response to your e-mail } a) The actual blasting unit that I use is relatively small in size. (2 ft } square) The volume of media used is around 3 quarts. In the eight years I } have been cleaning stubs the media has been changed maybe twice. This } material is treated as hazardous waste and disposed of accordingly. In my } opinion this is a small amount to dispose. Of course the relatively small } amount of stubs cleaned weekly (50 pcs) has a direct impact on media life. } I am not sure of most facilities volume, but if handled correctly } (dedicated blasting unit) this should not create problems. } b) That is a very good question. I honestly do not know of a procedure to } test to confirm the stubs are actually "clean" } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Many thanks for sharing that information with us. I am sure some of my } other } people will find it interesting. As I think I indicated, we have long } thought of a mechanism for "recycling" used mounts for people (many just } throw them out) but these other considerations have generally stopped us } dead in our tracks. But within a given laboratory, like yours, you would } pretty much know what was or was not on the mounts, and therefore would } know } how to properly protect yourself from exposure to bad things. } } } } Robert Fowler } Quality Assurance Technician (Failure Analysis) } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.tdk.com } } } } } "Monson, } } Frederick C." To: } "'robert.fowler-at-tdktca.com'" } {fmonson-at-wcupa {robert.fowler-at-tdktca.com} } } .edu} cc: } } Subject: RE: Aluminum Stubs } } 12/05/2001 } } 10:38 AM } } } } } } } } } } Morning Bob, } Would you mind telling me/us how you accomplish the sandblasting of } your used stubs. A real materials and methods paragraph would be most } helpful for us biologists in the audience. I will soon be surrounded by } bags of the things and I keep on finding more. Have tried sanding and } some } other methods, but all either mar the surface unsatisfactorily or leave } residue. } } Thanks in advance, } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } } } } ---------- } } From: "robert.fowler-at-tdktca.com"-at-sparc5.microscopy.com } } Sent: Tuesday, December 4, 2001 12:45 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Aluminum Stubs } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } Given that my curiosity is up per the plating/no plating of the stubs, } my } } posting is geared more towards the actual reusing of stubs. For the past } } eight years I have been subjecting a set of 50 or so stubs to a sand } } blasting treatment. You may question the feasibility of a sandblaster } BUT } } what is important is the media used to blast with. glass beads in the } } range } } of 170-325 mesh are used to effectively remove residue, glue, tape, and } } any } } material that resides on the stub. This particular media results in no } } residue and no tolerance change, although a pure water treatment and } } ultrasonic wash are utilized. I have had no trouble at all even after } this } } amount of time. } } } } Robert Fowler } } Quality Assurance Technician (Failure Analysis) } } TDK Components USA, Inc. } } Multilayer Ceramic Capacitor Division } } 1 TDK Boulevard } } Peachtree City GA 30269-2051 } } Telephone: (770) 631-0410 Ext.315 } } Fax: (770) 487-1460 } } email: rfowler-at-tdktca.com } } www.tdk.com } } } } } } } } } }
I apologize to all who recently received a queer email from me. I was hit by s new virus W32Aliz . Worm which my antivirus failed to detect and which is spreading in Microsoft Outlook Express without you open any atachment. Jiri Kalvoda
--- Odchozí zpráva neobsahuje viry. Zkontrolováno antivirovým systémem AVG (http://www.grisoft.cz). Verze: 6.0.306 / Virová báze: 166 - datum vydání: 4.12.2001
I apologize to all who recently received a queer email from me. I was hit by s new virus W32Aliz . Worm which my antivirus failed to detect and which is spreading in Microsoft Outlook Express without you open any atachment. Jiri Kalvoda
--- Odchozí zpráva neobsahuje viry. Zkontrolováno antivirovým systémem AVG (http://www.grisoft.cz). Verze: 6.0.306 / Virová báze: 166 - datum vydání: 4.12.2001
These are fundamental questions that you can learn about from a number of text books and courses. I would suggest starting with the SEM and Microanalysis book by Goldstein et al. You could also think about attending the Lehigh Microscopy Short Course which is very good and would teach you these things. David Joy has some Monte Carlo simulation software in the public domain that calculates these things and graphically displays them. You can get them at the EMMPDL - MASLIB - Mac & PC Shareware Libraries at Argonne National Laboratory. The following FTP Site give you access to these Software Libraries. Access is by anonymous Login:
Host : WWW.AMC.ANL.GOV or the mirror site WWW.MSA.Microscopy.Com
Login: Username = Anonymous Password = Your Email Address
You will have to hunt for them. Go to the main listing and look at the titles.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: ATC SEM Laboratory [mailto:atcsem-at-earthlink.net] Sent: Friday, December 07, 2001 8:09 AM To: Microscopy-at-sparc5.microscopy.com
Hello, I have been dealing with SEM/EDS for only 1.5 years and I have a lot to learn, but recently I have been asked a question: What is the EDS depth of penetration on my unit? I work with Amray 1645 and it was upgraded by PGT for imaging and EDS. Could anybody answer the question How deep does the x-ray beam penetrates through a sample and, if it is varies, how to find out actual depth?
If you have not lost cooling efficiency in the cold trap at the top of the diffusion pump or in the lines around the diffusion pump, I would wager that it is not diffusion pump oil. Check the cooling lines for adequate flow. If they are plugged, you could get diff pump fluid up in the chamber. Chances are that it is mechanical pump fluid. I trap will help, but you have to maintain it properly. To get rid of the oil in the chamber, there are some things that you could do. I recommend the Evactron system by XEI. I have checked it out (see the M&M 2001 proceedings) and it does clean up hydrocarbon oils in a vacuum chamber. XEI also has an overnight nitrogen purge system that helps clean microscope by using clean dry nitrogen. A colleague at one of our other research sites has this system on an older SEM and it really helps to keep beam contamination down. You could set up your own system with a leak valve and a pump that throttles the vacuum pump. You then keep the system in the viscous flow regime and it will clean the system. Better yet, you can put heaters on the tubing to heat the nitrogen going into this system and it will improve the "scrubbing" action of the gas molecules on the surface of the chamber.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Friday, December 07, 2001 8:42 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Dear Listers,
I have an older SEM which over a period of several months seems to repeatedly develop a small droplet of oil on the EDS detector collimator.
It is a diffusion pumped system. The mechanical backing pump is regularly serviced and gets oil changes.
I am inclined to think it is time to clean the diffusion pump and change the oil there. Perhaps over time backstreaming from the mechanical pump has occurred, causing contamination of the diffusion pump oil with mechanical pump oil. The latter, which has higher vapor pressure, is evaporating out of the diffusion pump into the chamber and condensing on the coldest point there (the detector).
Any opinions on whether this is reasonable or not? Any other possibilities anyone can think of? I'd hate to change the diffusion pump oil, which is going to be a big pain, if this is not going to do the trick.
By the way, I'm already planning to install a foreline trap (there currently isn't one).
Thanks,
Wharton
********************************************************** Wharton Sinkler UOP LLC Des Plaines, IL
Thanks for the note. I didn't take your remarks as disparaging - however, I did take advantage of them to have a little fun!
I now work for the NRC Institute for National Measurement Standards, which is the Canadian equivalent of your NPL, American's NIST, the "world's" BIPM, etc., so issues of traceability and ISO 17025 take up a lot of my time now. The fact that there is no officially traceable, high-magnification TEM calibration sample available has caused me (and probably most microscopists) some extra headaches. I maintain that TEM calibration samples based on crystal lattice spacing are fundamental constants of nature (i.e., "God -traceable") and hence do not require the blessing of a National Measurement Institute. It does occasionally take a bit of persuasion to convince the bureaucratically-minded that the whole point of traceability is not to refer to SI units through a National Measurement Institute, but to refer to nature through all of the above!
As you implied in your original posting, having a crystal as the basis of the sample allows self-calibration at better than the 1% level. Backing that measurement up with XRD is even more convincing. You have a nice advantage in your calibrated measurements in that you can lattice-match your alloy layers, and avoid worries about layer thickness variations as a result of strain. A potential problem with the SiGe marker layers in the MAG*I*CAL sample was that they were strained layers and hence had a slight variation in lattice parameter. We worked around that problem by self-calibrating the thickness of the individual SiGe layer(s)against the pure silicon crystal substrate. Silicon and TEM are a marriage made in heaven - the low atomic number of silicon makes the sample still useful for TEMs with accelerating voltages less than 200 keV. You are also correct about the robustness of Si relative to InP - in my experience, InP crystals will gratuitously cleave under any kind of stress, and usually in exactly the region of interest . . .!
Anyway, I appreciate the opportunity to discuss with you the subtlties of TEM calibration. The topic doesn't work it's way into social conversation very often. . .!
Cheers John
All the best.
Cheers John
John P. McCaffrey, Ph.D. Institute for National Measurement Standards National Research Council of Canada M-35 Montreal Road Labs Ottawa, Ontario K1A 0R6 CANADA
----- Original Message ----- } From: "Richard Beanland" {richard.beanland-at-marconi.com} To: "'John McCaffrey'" {john.mccaffrey-at-nrc.ca} Cc: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, December 07, 2001 5:13 AM
How are you preparing the Formvar?
Films should be freshly carbon coated. If not fresh, should be less than a week old--or if you're still having trouble with non-sticking, carbon coat same day). Also, you can "rejuvenate" them with glow discharge in a vacuum evaporator for 10-30 sec. You can also irratiate them with UV light for 30-40 min.
If neither is available, try poly-L-lysine (FW: 4000; 0.1% aqueous for a couple min, then wash in a couple drops of water); drain, dry). Other possible rx: alcian blue, benzyldimethyl alkylammonium chloride (BAC).
Hayat MA, Miller SE. Negative Staining. McGraw-Hill. 1990.
Sara Miller
On Fri, 7 Dec 2001 JHoffpa464-at-aol.com-at-sparc5.microscopy.com wrote:
} Date: Fri, 07 Dec 2001 08:18:49 EST } From: JHoffpa464-at-aol.com-at-sparc5.microscopy.com } To: microscopy-at-sparc5.microscopy.com } Subject: negative staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } hi all, i am doing collagen extractions from corneas and negative staing them to get the lenghts. my problem is that i am having trouble getting the collagen to stick to the formvar. any suggestions. i am on a deadline to work out the details of this project. } John } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Try chroma.com They are a great resource and have lots of information on their web site.
Dave Burton Optical Specialist University of Washington ----- Original Message ----- } From: {"ebhan-at-cybermed.ucsd.edu"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, December 06, 2001 2:32 PM
Dear Wharton and Microscopy List:
The assumption that your oil droplet is roughing pump oil distilled through the diffusion pump is very reasonable and is the most likely source. Michael Postek of NIST wrote a paper on this and a method he came up for control of the problem: "An Approach to the Reduction of Hydrocarbon Contamination in the Scanning Electron Microscope" Scanning Vol. 18, 269-274 (1996). He used a controlled leak into the foreline to stop backstreaming and control the problem. He also dicusses Cryotrps and Nitrogen purge systems as controls. I have reprints available of this article available for the asking. Send me your mailing address. Ric Felton found by IR that his oil drop was coming a oil outgassing from the Japanese Black Rubber vacuum hose supplied with his SEM. The oil can also come in from an airlock if the airlock is pumped continuously by a roughing pump.
I make my living by controling contamination and Oil backsteaming problems in electron microscopes. Doing business as XEI Scientific, I make the both a Nitrogen Purge system and the Evactron SEM-CLEAN Plasma Cleaning system for cleaning Electron Microscopes. Visit my web site at www. SEMCLEAN.COM for details.
Ronald Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 (650) 369-0133
-----Original Message----- } From: Sinkler, Wharton {WSinkler-at-uop.com} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com}
The Integrated Microscopy Core, Department of Molecular and Cellular Biology, Baylor College of Medicine has an immediate full-time opening for an electron microscopy technician. The Integrated Microscopy Core is a busy, state-of-the-art facility with a new Hitachi H7500 equipped with Gatan’s new 2kx2k CCD camera, deconvolution, laser scanning confocal, and 2 CCD-based upright and inverted epifluorescence microscopes. The applicant should have at least one year of experience in various aspects of sample preparation for biological TEM including fixation, embedding, ultrathin sectioning, staining and darkroom procedures. The applicant should have experience in the operation of TEMs and knowledge of computers. Other duties include preparation of solutions, embedding media and the maintaining of records. The position offers opportunities for training in advanced microscopy techniques, including digital TEM, fluorescence, deconvolution and laser scanning confocal microscopy. The position requires a minimum of a Bachelors degree and will start as a Lab Technician II; salary range is in the 30-40K, commensurate with experience, and the standard Baylor College of Medicine benefits package.
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I don't think this is a terribly unusual problem. I have operated two scopes with EDS detectors that have also developed oil drops on the end of the snout. The problem is that the collimator is chilled from the liquid nitrogen in the detector dewar, so any oil vapor in the chamber (and there is always some) goes right to it and condenses. If your scope is a variable pressure SEM one thing that helps is to maintain it at 7-10 Pa chamber pressure when not in use. This keeps a certain amount of gas circulating through the chamber and helps to flush out the backstreamed vapors. There is also a product called SEMCLEAN (I think), and I believe it operates on a similar principle.
Good luck, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Friday, December 07, 2001 7:42 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Dear Listers,
I have an older SEM which over a period of several months seems to repeatedly develop a small droplet of oil on the EDS detector collimator.
It is a diffusion pumped system. The mechanical backing pump is regularly serviced and gets oil changes.
I am inclined to think it is time to clean the diffusion pump and change the oil there. Perhaps over time backstreaming from the mechanical pump has occurred, causing contamination of the diffusion pump oil with mechanical pump oil. The latter, which has higher vapor pressure, is evaporating out of the diffusion pump into the chamber and condensing on the coldest point there (the detector).
Any opinions on whether this is reasonable or not? Any other possibilities anyone can think of? I'd hate to change the diffusion pump oil, which is going to be a big pain, if this is not going to do the trick.
By the way, I'm already planning to install a foreline trap (there currently isn't one).
Thanks,
Wharton
********************************************************** Wharton Sinkler UOP LLC Des Plaines, IL
At 09:03 AM 12/7/2001, you wrote: } Dear Gary, } } [snip]
} While I am not familiar with the Amray 1910, I have worked with many other FE } and conventional SEMs and I think your current measurements are not accurate. } In general, as beam currents increases, the spot size should also increase, } which is the opposite of what your data shows.
I would think that this SEM should exhibit the same characteristics as others would. I agree that the relationship of spot size and specimen current is whacky. I attribute this to my poor method of measuring the low specimen currents. As a check, I did the gun brightness test. It was working as normal. But the current value is probably 100 times what actual operating specimen current is. Gun brightness is measured without any final aperture and with condenser lens wide open (-35).
} I also have a graph from JEOL for the spot size vs. beam current for a JEOL } 840, which apparently is similar to what the other writer has. The graph I } have has almost a straight line on a log-log grid that goes through 10 nm at } 10 pA, 50 nm at 1 nA, and 500 nm at 500 nA (this is for a W filament at 30 kV } and the aperture size is increased as the beam current increases). } Unfortunately, the JEOL data does not indicate how the "spot diameter" was } measured.
Assuming we use the same "sharp corner" method, the log-log data means that the relationship is not linear. My data suggests that it is linear. So again, my measurements are suspect. The only thing I seem to find believable about what I measured is that spot size gets smaller at higher CL settings. At least this is a good correlation direction.
} Spot size vs. beam current data from an older Philips FE SEM at 10 kV with a } 30 um aperture is as follows: ~0.5 nm at ~10 pA, ~3.5 nm at ~0.55 nA, and } ~30 nm at ~35 nA. } } It is also interesting that in your data, over the full range of CL settings, } the currents you have measured hardly change, while most SEMs will have a } full range of several orders of magnitude or more. } } If possible, I would suggest you borrow a Keithley 485 picoammeter (or } similar) to check the current measurements you have made. Note that if the } sign of the measurement changes at low beam currents, it indicates that there } is a grounding problem that has a leakage current that is larger (and } opposite) of what you are measuring. Also note, that on a picoammeter like } the 485, the displayed reading when connected to a Faraday cup in an SEM will } always be negative when working correctly.
I ordered a 485. It should be here in two weeks. I'll re-do all measurements when it gets here. Any special notes I should know about when using it? It has a BNC input jack so I'll just coax from it to my stage alarm BNC.
What could I do to check the stage grounding situation? The BNC alarm output is not grounded (shorted). Perhaps I should check the BNC ground side to be sure it is grounded. That would mess things up for sure. But the gun brightness measurement would not have worked correctly.
} My background is in using electron microscopes for e-beam lithography. In } that case, the exposed spot is always larger than the size of the incident } beam. This results from the fact that as soon as the beam hits, there will } be scattering and it is the secondary electrons that have the highest } probability for exposing the resist. For comparison, a high performance FE } SEM at 30 kV and ~20 pA of current can expose lines down to 10 or 20 nm, } which puts an upper limit on the beam size for those models at those } conditions. Since such models have a resolution spec of 1 to 2 nm above 10 } kV, the lithography size is reasonable, since the resolution measurements are } not limited by a resist resolution or by broadening by secondary electrons.
We use Heidelberg DWL to write masks. The laser makes patterns down to about 0.1u which is all we need at 0.25u feature size. It is probably better than that but I have never measured it.
} In general, the approach for measuring resolution is to see how small a gap } the beam can fit between islands of gold. While this approach does not give } any information on the profile of the beam as you are trying to measure, it } certainly gives reasonable data on the "width" of the beam. Quite simply, if } the image is dark when the beam is positioned in a 5 nm gap between to gold } islands on carbon, then it is clear that "most" of the beam is hitting the } carbon and not the gold. It would be interesting if you could correlate your } width measurements to values from a resolution measurement.
there might be another approach to performing a sanity check. I can set up some IC process control monitors which have various geometries of metal fingers meshed between one another. Each set of fingers (E with lots of fingers sticking out) are meshed together but not touching. With 24VDC or 12VDC in the chamber for the stepper motors and limit switches, I could connect one E to +12 and bring the other E out through the picoammeter and place a beam at various points. This ought to cause current flow when the beam transcends two adjacent fingers. Might be worth a shot.
I am still searching for a true sharp corner. They are not all that sharp. At 100KX and above, the detail in their surface shows up and degrades a sharp delineation at the boundary.
Gary G.
} Joe } _________________________________________ } Joe Nabity, Ph.D. } JC Nabity Lithography Systems } E-Beam Lithography using Commercial SEMs & STEMs } PO Box 5354, Bozeman, MT 59717 USA } Voice: (406) 587-0848 } FAX: (406) 586-9514 } E-mail: info-at-jcnabity.com } Web: www.jcnabity.com
This is something we had been experiencing with our SEM and our former beryllium window detector. The X-ray detector had to be removed more than once to wash oil off the window, in spite of having upgrading the diff pump oil to Santovac to try to minimise the contamination. I analysed some of the oil deposited on the collimator by FT-IR and it wasn't Santovac oil, it was backstreaming diff pump oil. Then when we upgraded the X-ray system a couple of years ago to a thin (polymer) window detector, I decided something had to be done about the oil, even though the new detector surfaces weren't so cool as the previous one's.
One problem with foreline traps is that they slow down pumping speed. So I cannibalised a much bigger rotary pump from an old mass spec that was being binned, had it reconditioned, fitted a good foreline trap and haven't looked back. If you can do the same, I'm sure you will be pleased with the effort.
"Sinkler, Wharton" {WSinkler-at-uop.com} on 07/12/2001 13:41:57
To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} cc:
Dear Listers,
I have an older SEM which over a period of several months seems to repeatedly develop a small droplet of oil on the EDS detector collimator.
It is a diffusion pumped system. The mechanical backing pump is regularly serviced and gets oil changes.
I am inclined to think it is time to clean the diffusion pump and change the oil there. Perhaps over time backstreaming from the mechanical pump has occurred, causing contamination of the diffusion pump oil with mechanical pump oil. The latter, which has higher vapor pressure, is evaporating out of the diffusion pump into the chamber and condensing on the coldest point there (the detector).
Any opinions on whether this is reasonable or not? Any other possibilities anyone can think of? I'd hate to change the diffusion pump oil, which is going to be a big pain, if this is not going to do the trick.
By the way, I'm already planning to install a foreline trap (there currently isn't one).
Thanks,
Wharton
********************************************************** Wharton Sinkler UOP LLC Des Plaines, IL
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When I get a question about SEM/EDS analysis depth, I refer people to the ASTM E 1508-98 "Standard Guide for Quantitative Analysis by Energy Dispersive Spectroscopy" which is in volume 3.01 if you have the annual books ASTM publishes. In paragraph 8.2.3, there is a rule of thumb to estimate the analysis depth where 99% of the x-rays are emitted. The rest of the guide is also a useful, brief, reference to be familiar with when doing EDS experiments.
Regards, Dave Audette david.audette-at-sylvania.com
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Soda glass, is used in most and especially the cheaper microscope slides. Soda glass does fluoresce. If you look at the slide side on and the glass has any tinge of green, than its soda glass. If you can, just place a couple of coverslips on the stage - hopefully you would not get fluorescence, proving its the slide. The slides to use for any fluorescence applications are the water- white or superwhite slides. These the manufacturer claims, do not fluoresce at all - probably true in normal microscopy situations, but one customer with an extra powerful detection system managed to get some slide fluorescence from the superwhite slides too. Disclaimer: ProSciTech supplies Superwhite slides. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, December 08, 2001 12:15 AM, James Pawley [SMTP:jbpawley-at-facstaff.wisc.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Russell, } } If the pattern is the same, the signal is probably still reflected } light. This signal can be very large compared to fluorescence and it } is just blasting through (or around) the filters/prism. } } Jim Pawley } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Sorry about the subject name, it is a bit misleading. In } } using a Leica SP2 } } confocal, I have noticed that it is possible to see the cover slip } } and slide. Why/how is this possible? The setup is essentially this: } } a coverslip is placed } } on a slide, which is then placed on the stage (there is nothing between them, } } } } except of course for miscellaneous small dust particles and air). The slide } } is } } then scanned with light from a HeNe laser (633nm). When the light detection } } (wavelengths directed to the PMT's) is set to include 633nm (i.e. } } 610-650nm), the } } signal is of course very bright. I am assuming that this is due to } } reflection. What I am confused about is that if the light detection } } doesn't include 633nm } } (sampling from 660-710nm), there is still signal present, and it is } } in the same } } pattern as is seen when looking at the reflected light. It is } } important to note } } that this pattern is connected to the coverslip and slide somehow, } } as moving the } } stage will cause a corresponding change in the image produced by the system. } } I realize that I may be missing something fairly obvious, but } } any help would } } be greatly appreciated. } } } } Thanks, } } Russell } } } } } } } } -- } } Confocal Imaging Facility Technician } } Tufts Medical School } } 136 Harrison Ave. } } Boston, MA 02111 } } Tel. (617) 636-3795 } } -- } **************************************** } Prof. James B. Pawley, Ph. 608-263-3147 } Room 223, Zoology Research Building, FAX 608-265-5315 } 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU } "A scientist is not one who can answer questions } but one who can question answers." } Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I am an art conservator working in Japan and would like to purchase a pigment reference set and mounting medium but have been unsuccessful in my web searches.
I seem to recall that McCrone and Fisher sold such pigment reference sets--with about 40 or so mounted samples of common artist pigments, but their web pages did not turn up with any such item.
Also, I would like to know what is the best refractive index for medium used to mount pigments? Is is 1.55? When I was in the U.S. I used Cargille mounting medium.
Thank you so much for your assistance, since it is quite difficult for me to find information on this in Japan.
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Dear Wharton, It seems like all of the responses to your inquiry are trying to solve the effect of your problem rather than the cause. The oil that is condensing is most likely mechanical pump oil if your vacuum system is working properly. In any vacuum system using a oil charged mechanical pump and a diffusion pump there will be oil vapors. These oil vapors are also the cause of contamination spots on the sample after the beam has stayed in one location for a period of time. Unless you upgrade your vacuum system to a dry mechanical pump and a turbo molecular pump, oil vapors will be present. Now the cause of your problem is the EDX detector itself. EDX detectors are static vacuum systems. They are initially pumped out; when LN2 is present in the dewar, which is attached to a cold finger holding all of the internal components, cryostatic pumping occurs within the EDX detector. With a well maintained EDX detector, very little thermal loss is transferred to the external portion of the detector. In your case, the internal vacuum of your EDX detector is poor and a high thermal loss causes the EDX tube to be cold and therefore becoming coldfinger within the vacuum system. This coldfinger ( your EDX tube) is now attracting the oil vapor present in your SEM and creating the oil droplets. To solve your problem you need to have a vacuum treatment ( pump-out and bake-out )performed on your EDX detector. If you would like additional information concerning EDX detectors, please visit my web site xraydetectors.com. Regards, Jim Nicolino
"Sinkler, Wharton" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } I have an older SEM which over a period of several months seems to } repeatedly develop a small droplet of oil on the EDS detector collimator. } } It is a diffusion pumped system. The mechanical backing pump is regularly } serviced and gets oil changes. } } I am inclined to think it is time to clean the diffusion pump and change the } oil there. Perhaps over time backstreaming from the mechanical pump has } occurred, causing contamination of the diffusion pump oil with mechanical } pump oil. The latter, which has higher vapor pressure, is evaporating out } of the diffusion pump into the chamber and condensing on the coldest point } there (the detector). } } Any opinions on whether this is reasonable or not? Any other possibilities } anyone can think of? I'd hate to change the diffusion pump oil, which is } going to be a big pain, if this is not going to do the trick. } } By the way, I'm already planning to install a foreline trap (there currently } isn't one). } } Thanks, } } Wharton } } ********************************************************** } Wharton Sinkler } UOP LLC } Des Plaines, IL
"Sinkler, Wharton" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } I have an older SEM which over a period of several months seems to } repeatedly develop a small droplet of oil on the EDS detector collimator. } } It is a diffusion pumped system. The mechanical backing pump is regularly } serviced and gets oil changes. } } I am inclined to think it is time to clean the diffusion pump and change the } oil there. Perhaps over time backstreaming from the mechanical pump has } occurred, causing contamination of the diffusion pump oil with mechanical } pump oil. The latter, which has higher vapor pressure, is evaporating out } of the diffusion pump into the chamber and condensing on the coldest point } there (the detector). } } Any opinions on whether this is reasonable or not? Any other possibilities } anyone can think of? I'd hate to change the diffusion pump oil, which is } going to be a big pain, if this is not going to do the trick. } } By the way, I'm already planning to install a foreline trap (there currently } isn't one). } } Thanks, } } Wharton } } ********************************************************** } Wharton Sinkler } UOP LLC } Des Plaines, IL
Apologies. I have just reread my message and can see the mistake. The analysed oil that had condensed on our detector collimator did not give the characteristic FT-IR spectrum of a polyphenol ether, e.g. Santovac oil, it was rotary pump oil (and not diff pump oil). Hence, a foreline trap was fitted to the rotary pump.
"Garber, Charles A." {cgarber-at-2spi.com} on 08/12/2001 05:19:15
To: "jill.webb-at-rssl.com" {jill.webb-at-rssl.com} cc:
-------- REPLY, End of original message --------
******************************************************************** This e-mail is confidential and may contain privileged information. If you are not the addressee it may be unlawful for you to read, copy, distribute, disclose or otherwise use the information in this e-mail. If you are not the intended recipient please notify us immediately.
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My FESEM is Schottky ZrOW, thermally assisted. I believe that all Hitachi FESEMs are cold cathode. If so, my experience probably does not apply.
The Schottky filaments have to be "conditioned" prior to use. This process involves having the gun assembly sitting by itself and braid strap grounded to Earth ground. The entire gun electrode set is then brought up to 32KV while a separate gun chamber ion pump power supply is connected and monitored for pump current.
The potential is raised slowly, about 500V per 5 minutes or so. The pump current is constantly watched. If it starts to increase, the potential is reduced, let sit and then raised again. At some point, the unit may zap. This drive the pump current way high and the whole process starts over again. Once the gun assembly can sit at 32KV for an hour with low pump current and no zapping, it is then mounted on the column. It should never zap again during operation at 30KV max rated potential.
When connected to the column and console electronics, a zap will most likely take out some electronics in the system. This is why the gun is conditioned off of the column.
I don't have an emission current control. The only computer controlled setup values are for extractor voltage and suppressor voltage. Once set, they are not adjusted again under normal operation.
I would think that any zapping is an indication of dirt or crud on the gun area. What does Hitachi say about this?
gary g.
At 08:44 AM 12/5/2001, you wrote:
} Dear listers, } } This query is directed specifically at users of the Hitachi S4700 FESEM, but } may also apply more widely to users of FESEM's. } } We get a curious result (to me) when flashing the tip. We were advised to } flash until the emission current reading read about 20uA on the "le" meter. } The flash intensity adjustment behind the scope was initially set to } accomplish this in one or two flashes when flashing at an intensity of "2" } in the Setup-Column menu. } } That is what happened for quite some time, however for a while now flashing } has resulted in values between 7-12 on the "le" meter. The initial flash } briefly degrades the vacuum at the gun as expected as contamination is } driven off, but subsequent flashes do not, also as expected. However, } subsequent flashes usually LOWER the "le" meter value, rather than raise it } toward our goal of 20. This is the really puzzling part to me. Originally, } the value increased with each flash, and if an excessive number of flashes } were required, we would adjust the intensity pot in the rear of the scope. } } We normally run the scope at an emission of 10uA, with kV's of 1.0-10.0. } All vacuum readings are normal, and the scope performs beautifully in terms } of resolution and brightness. } } My questions are: } } 1) Why is the "le" value decreasing with multiple flashes? } } 2) How dangerous are multiple flashes to the tip? } } 3) Does the condition described above indicate any problem with the tip } and/or scope? } } 4) Is flashing actually accomplishing anything if the vacuum gauge shows no } contamination being driven off? } } 5) Is it safe to keep turning up the intensity pot on the rear of the } scope? (Something tells me no.) } } 6) Do I simply not understand what's going on? (Something tells me yes.) } } Thanks in advance! } } Puzzled in Missouri, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/
I saved an old Cook CV-301 thermal evaporator from another group from a dumpster and I rebuilt it. It has two sets of boat holders mounted on the base plate. The boat gap is 3" and the clamping screws have 3/16" diameter threads. The bell jar is 12" in diameter by 12" high. Pump down pressure was improved from 10E-3 to 3 - 5 x10E-6, now. I have no accessories for the thing. I have no manuals (hint), no shutter, etc.
I want to set this up as a dedicated old style 1/8" carbon rod and a W basket (mgs of metal) evaporator. I buy my TEM grids but sometimes they are too light on carbon or I would like to increase the conductivity of the carbon 'back coated' formvar grids. Thus, I would like to 'thicken' them up a bit with carbon. I do NOT want to send big 'boulders' of carbon smashing through perfect featureless grid films, however.
So I need to buy a carbon evaporation unit of some type. Should I go with an old style combo unit like Ladd Research sells? Many think carbon fiber / cord / strings are the way to go and the Ladd unit does not appear to do that type of C. Obviously, I would want to have a W basket or W wire setup as a option for my troubles saving this unit.
Ladd sells both combination and separate units to do both jobs and they can be shielded. Have you used one of these? Fullams sells much cheaper priced units, but are unshielded. Have you used them on a non-EFFA evaporator or port? Does the carbon rod spring work smoothly and reliably on any of these units?
What other after market units are reliable based on your REAL experiences and what would you buy today, if you had to do it over again, like me?
One final, low priority, question on DP oil: The Cook is simpler to use than my Edwards 306A but it only has a 400-450 watt DP heater on a Varian HS2 DP. It calls for 100mls of Dow Corning 704 silicone oil. Not my first choice. I think the heater is too small in wattage for Santovac based on my experience with a 1964 JEOL JEM 7 TEM oil conversion attempt. I could try to switch to another DP fluid like Apiezon A, B or a Fomblin. DC 704 boils at about 215°C. So, have you ever done this type of oil conversion successfully and what oil did you use? Alternatively, I also have a degree in Electronic Technology with honors of the highest distinction and could take the controling circuit board out, figure out the schematic and convert the design to supply a 1KW or larger heater. However, I am not sure this HS2 DP can handle a 500 - 600 watt heater using Santovac for the next 10 years I have to work. Notes: The HS2 DP seems to be a bit too slow in pumping speed to me and might be undersized. It is not the original DP unit but looks right mechanically except for a slight roughing port mis-aligment of 1/4 " at the 4" long vac. hose outlet. The chimney is clean, aligned and assembled correctly.
Thank you in advance for any PRACTICAL experience(s) you can provide. I am trying to avoid the trial and error route.
Sincerely,
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center Monroeville, PA 15146 (724) 325-5131
You are correct. The process you describe does not apply to the Hitachi S-4700 FESEM.
The Hitachi S-4700 is a cold FE. The procedure you describe is for the Amray "305" gun which is normally brought to 35 KV for one hour or more. If the engineer tells you 32KV, he is probably cutiing corners.
For the Amray 305 gun: The zapping you describe is from stray molecules that have a tendency collect randomly around the gun, including the insulators. I have found that there are several "thresh-holds": voltages at which the gun arcs. These are proportional to vacuum integrity. The "thresh-holds" are at 14-15 KV, 24KV and 30KV. The 305 guns will arc then the vacuum will need to be "cleaned up" by gettering. We then start the whole conditioning process over again.
Although the indicated vacuum on the Amray CRT reads about 10e-10 Torr, the actual pressure is closer to 1x10e-8 or 8x10e-9. I have actually measured this from a calibrated hot cathode gauge.
Amray guns could use with more assistance from an additional ion pump. The existing ion pump is rated at 2 l/s. This is barely enough to "hold" the vacuum & seems to be nothing more than an "indicator" gauge. I have installed a 20 l/s ion pump on the gun and have had considerable success with less arcing, faster pump down times, better resolution, etc. It is no mistake that most other Schottky FE guns have this arrangement (LEO, Hitachi, JEOL).
Now for the Hitachi 4700 Cold FESEM questions:
} 1) Why is the "le" value decreasing with multiple flashes?
It shouldn't. Most Hitachi guns I have seen always increase with multiple flashes. I would suspect contamination or a gun problem.
} 2) How dangerous are multiple flashes to the tip?
Multiple flashes have a tendency to "blunt" or "round" the tip giving the tip less filament life but more filament stability. Hitachi actually recommends multiple flashes to condition a new tip.
} 3) Does the condition described above indicate any problem with the tip } and/or scope?
See the answer to question #1.
} 4) Is flashing actually accomplishing anything if the vacuum gauge shows no } contamination being driven off?
The first flash sould drive off any molecules at the FE tip and, therefore, give a poor, but temporary, vacuum indication. Successive flashes should affect the FE tip but will not change the vacuum.
} 5) Is it safe to keep turning up the intensity pot on the rear of the } scope? (Something tells me no.)
Define "safe" and the amount you are turning up the intensity pot. Hitachi typically has three flash intensity settings (approprately named: 1,2,3) with "3" being the most intense. I usually set "1" for 5 ua, "2" for 10 ua and "3" for 30 ua on the FIRST flash. Sucessive flashes have a tendency to increase the emission current. This is rather tricky as each filament will have a different setting. I would guess that anything over 40 & above is probably melting the FE tip and giving you poorer filament life.
} 6) Do I simply not understand what's going on? (Something tells me yes.)
Based on the depth and type of questions you ask, I think that you do know what is going on. I also would be concerned also if sucessive flashes decrease the emission current.
Summary: my guess is that there is something wrong (or going wrong) in the gun area.
What is the extraction voltages before& after all this?
DISCLAIMER: THE ABOVE ARE BASED UPON MY EXPERIENCE & ARE OF MY OPINION AND OF MY COMPANY, SCANSERVICE CORP. TO THOSE THAT WOULD CRITICIZE THE ABOVE BASED UPON A MIS-SPELLED OR MISPLACED WORD (especially from acadamia) I WOULD ONLY SAY "GET A LIFE".
Any and all CONSTRUCTIVE advice is certainly welcomed.
Regards,
Earl Weltmer
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Tindall, Randy D." {TindallR-at-missouri.edu} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, December 09, 2001 1:56 PM
I have some samples of tungsten carbide and diamond dust in mineral oil taken from grinding machines. How do I separate these from the oil for LM and SEM and disperse them for examination? I need to get these particles into a solvent so that the particles separate. Oil makes the particles clump together. Carbides are heavy and sharp and stick to the sides of all containers so it is just too easy to get a false picture.
Many thanks. Gervais Sawyer. Forest Products Research Centre. UK.
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Although there are many excellent books on histochemical techniques for animal tissues, I have been unable to find one still in print that concentrates on plant tissue. I would appreciate any recommendations so I can add this resource to our facility mini-library.
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA16619 for dist-Microscopy; Mon, 10 Dec 2001 10:38:39 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA16615 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 10 Dec 2001 10:38:08 -0600 (CST) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA16608 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 10 Dec 2001 10:37:57 -0600 (CST) Received: from NPGSlithography-at-aol.com by imo-m01.mx.aol.com (mail_out_v31_r1.9.) id 6.126.89d294b (3699); Mon, 10 Dec 2001 11:34:10 -0500 (EST) Message-ID: {126.89d294b.29463e01-at-aol.com}
Dear Gary,
} I ordered a 485. It should be here in two weeks. I'll re-do all } measurements } when it gets here. Any special notes I should know about when using it? } It has a BNC input jack so I'll just coax from it to my stage alarm BNC.
First, the stage alarm circuit must not be connected while using the picoammeter. If the BNC is normally connected to the alarm circuit, removing the cable to the alarm and connecting the picoammeter should be fine. The stage should have no other grounding path other than through the picoammeter.
(Note that the stage alarm will then be disabled, so it shouldn't be left this way if inexperienced users will be running the SEM.)
Ideally, the power outlet used by the picoammeter will have the same grounding as the SEM.
A good reality check is to connect the picoammeter and take a reading while the SEM is powered up, but the beam is off. With good grounding, a reading of zero will result. If a non-zero reading is measured, a quick and dirty solution is to use the "relative" mode of the Keithley 485 in order to take subsequent readings relative to the beam off baseline. This approach assumes that any leakage current being measured is a constant. Valid beam current measurements from a Faraday cup will always be displayed as negative numbers with the Keithley.
Also, note that a standard BNC cable will generate tens or hundreds of picoamps through internal friction if it is flexed or even bumped. Consequently, a relatively short BNC cable that is reasonably motionless will give the best measurements.
Finally, the Keithley 485 has a local magnetic field that is significant compared to the typical environmental specs for an SEM. Consequently, for best imaging, the 485 should be more than ~1 meter from the column. It is especially important to note that the 485 has a higher local magnetic field when the front power button is in the off position, than when the meter is turned on. Consequently, if the meter is close to the column, it must be unplugged to stop the field.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Thanks to everyone who replied to my questions about flashing the tip in our FESEM giving us odd flashing current readings. I received some very informative and helpful replies, as always from this list.
Incidentally, I didn't put the question to the list because of any lack of helpfulness by Hitachi Instruments. I was just curious about the experiences of fellow users and what they thought about the theory behind this. We love this scope!
Happy Holidays, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
While Jim Nicolino is says about EDS detectors acting as cold fingers can be true. However, I wish to point out that not all oil diffusion pumped are SEMs are the same as far as oiliness. For example, old Hitachi S-570s are infamous for being oily because they lacked a proper baffle above the diffusion pump mouth. I have never cleaned a S-570 without finding oil dripping from the bottom of the baffle plate at the bottom of the chamber. Hitachi S-2500 and S-2700 which updated the S-570s did not have as severe of a oil problem because of an improvement to the design.
Also contributing to the effect are choices of pump oil, heater temperature, cooling water temperature, and crossover pressure. If any of these is wrong then their will be severe backstreaming from the diffusion pump. Will Biglow's book "Vacuum Methods in Electron Microscopy" is good reference.
In modern JEOL and Hitachi SEMs that have diffusion pumps rarely does one find that it is the diffusion pump giving the problem. More common is oil backstreaming from the load locks. Many a user has replaced the diff pump with a turbo pump only to get little improvement because of the rotary oil roughing pumps, poor sample handling, and oily chambers and stages.
Notice: XEI Scientific makes products to remove oil from SEM chambers. www.SEMCLEAN.com
Ronald Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 (650) 369-0133
-----Original Message----- } From: Jim Nicolino {JNicolino-at-fcol.com} To: Sinkler, Wharton {WSinkler-at-uop.com} Cc: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com}
McCrone continues to sell the small set of reference pigments. Perhaps you did a search on the "McCrone Associates, Inc." page instead of on the "McCrone Microscopes and Accessories" pages.
The direct link to the page is:
http://www.mccrone.com/cgi-bin/SoftCart.exe/mac/reference/paintref.html?L+mccrone+svuu9375+1007930922 The choice of mounting medium depends on several factors related to your own work. It is often convenient to have a mounting medium with higher refractive index somewhere in the middle of the range of refractive indices of your samples. First, in many cases a rapid subdivision between pigment types can be made on the basis of whether they are above or below the r.i. of the mounting medium.
Second, pigments tend to have high refractive indices for the simple reason that, in order to function well as pigments, they must be effective at scattering light. The greater the difference in r.i. between the organic binder and the pigment, the more effective the scattering and the greater the opacity of the paint. If you chose a medium with a low r.i., the large index difference between the particles and the medium will make it more difficult to observe the optical properties and shape of very fine particles.
In general, Far Eastern art done in aqueous media makes greater use of low r.i. pigments in the form of clays, micaceous minerals and so on. So the nature of the art which you spend time on is something to consider as well.
The softening temperature of the mounting medium is a factor to consider. It is convenient to have a material which remains viscous near room temperature so that particles may be observed while rolling under the cover slip. On the other hand, if you store mounted samples for long periods, such media will cold-flow and require you to store your slide cases standing on end so that the slides remain horizontal to keep the coverslips from sliding off.
Finally, I would caution against placing too great a confidence in pigment identifications based primarily on comparison to a limited reference set. Many conservation studies suffer from erroneous pigment identifications based PLM comparison to the most similar example in a limited reference set rather than an actual identification. There is no substitute for instrumental analysis to supplement microscopy.
Disclaimer: I am not associated with McCrone A. & C. and have no interest the company.
John Twilley Art Conservation Scientist
c. yeh wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } I am an art conservator working in Japan and would like to purchase a } pigment reference set and mounting medium but have been unsuccessful } in my web searches. } } I seem to recall that McCrone and Fisher sold such pigment reference } sets--with about 40 or so mounted samples of common artist pigments, } but their web pages did not turn up with any such item. } } Also, I would like to know what is the best refractive index for medium } used to mount pigments? Is is 1.55? When I was in the U.S. I used Cargille } mounting medium. } } Thank you so much for your assistance, since it is quite difficult for } me to find information on this in Japan. } } } __________________________________________________ } FREE voicemail, email, and fax...all in one place. } Sign Up Now! http://www.onebox.com } } } }
I recently acquired an AMRAY 1610T with PGT 4000 EDS. (Thanks, Melissa!)
I hope to restore it to working condition in my spare time. The manuals that were included appear to be incomplete copies. For example, the SEM manual doesn't include schematics, but the vacuum logic manual does.
Are there also service manuals for this? What might be my best route to getting a set of complete manuals?
Here's a question - can joysticks be serviced? The one that controls the stage movement on our ElectroScan E3 has been getting a little wonky lately; movement is quite jerky, especially in side-to-side motion. I'm thinking maybe there's a build-up of crud or something in there that might be causing this. There are four small Phillips-head screws that seem to hold the joystick in place on the control panel, and I wonder if we could get at the mechanism that way. Has anyone out there ever worked on their own joystick? (There's just no way to say that without it sounding kind of funny...)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
We have a small room for cryoesctioning and storage of a liquid nitrogen container. For the safety concern, I would like to get a oxygen sensor or alarm to monitor the oxygen level. Does anyone konw where I can get this kind of device? Any suggestion will be appreciated. Thanks in advance.
Shanling
Shanling Shi Advanced Imaging & Measurement Unilever Research US 45 River Road Edgewater, NJ 07020 201-840-2340 Shanling.Shi-at-unilever.com
This AirAware model is AC powered (Comes with an AC adaptor, no batteries to worry about.), simple to install, and has a loud alarm. The sensor is relatively inexpensive and easy to replace. We use it in our freeze fracture/cryo microtome room.
Good Luck, Jim
Disclaimer: We have no monitary interest in PriorityOne or Lab Safety Supply.
} } Hello everyone, } } We have a small room for cryoesctioning and storage of a liquid nitrogen } container. For the safety concern, I would like to get a oxygen sensor or } alarm } to monitor the oxygen level. Does anyone konw where I can get this kind of } device? Any suggestion will be appreciated. Thanks in advance. } } Shanling } } Shanling Shi } Advanced Imaging & Measurement } Unilever Research US } 45 River Road } Edgewater, NJ 07020 } 201-840-2340 } Shanling.Shi-at-unilever.com }
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 Storrs, CT 06269-2242 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
Does it have an ion pump on the gun chamber? If so, it is set up to handle LaB6, otherwise W only.
What seems to be the problem with the system at present? Is it connected and powered? Check all vacuum connections. A good option is to remove the standpipe (if ion pump is present) and replace with Lesker butterfly valve and serrated stainless steel flex pipe with KF fittings. The standard vertical stand pipe can be a a leaker.
The UVACOS vacuum board is pretty much standard for all 1000 series SEMs. The 1610T added the Balzers 240 turbo and controller. The slow scan nature of this system makes it difficult to use. I brought a 1610T to life about four years ago. I moved on to an 1830 and and now have a 1910FESEM. There is much similarity between these extremes of models.
If there are specific sections you need, I have a complete manual for AMR1000.
gary
At 10:23 AM 12/10/2001, you wrote:
} I recently acquired an AMRAY 1610T with PGT 4000 EDS. } (Thanks, Melissa!) } } I hope to restore it to working condition in my } spare time. The manuals that were included appear to } be incomplete copies. For example, the SEM manual } doesn't include schematics, but the vacuum logic } manual does. } } Are there also service manuals for this? What might } be my best route to getting a set of complete manuals? } } - John
Announcing the Seventh Annual INTERNATIONAL 11-Day Short Course on
3D Microscopy of Living Cells, June 10 - 20, 2002
Sixth, Post-course Workshop on, 3D Image Processing, June 22 - 24, 2002
Organized by Prof. James Pawley, University of Wisconsin-Madison
in association with the, UBC Brain Research Centre, Prof. Max Cynader, Director. University of British Columbia, Vancouver, BC, Canada
(CORRECTION!! some early brochures were sent out with an incorrect URL. Course info can be found at: ht tp://www.3dcourse.ubc.ca )
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, the organizers have designed an intensive eleven-day residential course concentrating on all aspects of the 3D Microscopy of Living Cells. Sponsored by the Brain Research Centre at the University of British Columbia, it will be held in June of 2002. The course includes 4 days on 2D techniques, 5 days of 3D techniques and 2 days on 3D measurement and display. It includes everything from basic microscopy to confocal and multiphoton microscopy. A half-day Pre-course is offered for those wishing to brush up on (very!) basic optics.
Topics include: o Quantitative 2D light microscopy o 3D Imaging in confocal and widefield o Fluorescent and backscattered light signals o Digitization: The Nyquist Criterion o Poisson noise QE and S/N. o Lasers and laser tweezers o Objectives and aberration correction o Scanning-systems: AODs, mirrors, disks o Wide field/deconvolution techniques o Detectors: operation and performance o Optimal pinhole size/photon efficiency o Dye design, characteristics and use o How to keep your cells alive o Multi-photon excitation o Measuring ion concentrations o Display and measurement of 3D data
Morning lecture/demonstrations lead to hands-on laboratory exercises each afternoon utilizing most of the commercial instruments currently available for 3D microscopic imaging. Students will work in groups of 3 or 4 throughout the discussion and laboratory sessions, and may complete a live-cell 3D study on their own specimens. In the first 6 years, over 160 students from 25 countries have attended. Last year, 12 separate 3D microscopical workstations were each scheduled for over 30 hours of student use under the supervision of an international faculty of 17 who represent the state-of-the-art in live-cell microscopy. Including manufacturers representatives, the teacher/student ratio will be almost 2:1.
INTERNATIONAL FACULTY o Stephen Adams University of California-SD o Dan Axelrod University of Michigan o Mark Cannell University of Auckland o Milton Charlton University of Toronto o Ping Chin Cheng SUNY, Buffalo o Stefan Hell Max Planck Institute, Goettingen o Alan Hibbs BioCon, Melbourne, Australia o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andres Kriete Tissue Informatics, Pittsburgh o Glen MacDonald Virginia Bloedel Hearing Inst, WA o Irina Majoul Max Planck Institute, Goettingen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Technology o Chip Shook Eppendorf Scientific,Albequerque, NM o Jim Turner Wadsworth Institute, NY o Michael Weis Agriculture Canada
TUITION Course tuition is $2,250 US and includes lunches and generous snacks. On receipt of 50% deposit, all students will receive preliminary group assignments and a copy of the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes the Opening Reception, the Manufacturer's Reception, and the Beach Party, and a handout binder. Accommodations and other meals are not included. The Pre-course is $100 US
APPLICATIONS Applicants will submit an application to assess knowledge level and field of interest. Enrollment will be limited to 24 - 32 participants (depending on equipment availability). Selection will be made on the basis of background and perceived need. Those with little previous LM experience will be provided with basic texts to read before the course begins, and should take the Pre-course.
Application forms and other course information from this and past years can be downloaded from the WWW site at
h ttp://www.3dcourse.ubc.ca/home.html
or obtained from:
Prof. James Pawley, Zoology Department., 1117 W. Johnson Drive, Madison, WI. Phone: 608-263-3147 fax. 608-265-5315 Email: jbpawley-at-facstaff.wisc.edu
IMPORTANT DATES Applications must be received by Mar. 15, 2002 Deposit due Apr. 15, 2002 Registration 5:00 - 7:00 pm Sunday, June 9, 2002 Intro. Lecture 7:00 PM, Sunday, June 9, 2002 Last class ends with lunch, Thursday, June 20, 2002
-- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I have a query from a collegue carrying out autoradiography of an uneven surfaced structure. She has found that standard autoradiographic plates are not suitable and is interested in using a liquid emulsion or stripping film. I have directed her towards Ilford as a source of a liquid emulsion - has anyone any other suggestion and is stripping film (Kodak AR 10) still available.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
we are using a light microscope to acquire brightfield images of diatoms, which we then want to process by computer. However, the presence of diffraction effects around the edges of the images is causing us problems.
Does anyone know of any software, preferably free, which can deconvolve an image to reduce the effects of diffraction?
thank you
Micha Bayer ______________________________
Dr. Micha Bayer Royal Botanic Garden Edinburgh 20A Inverleith Row Edinburgh EH3 5LR Scotland, U.K. Tel. (+44) (0)131-248 2915 Fax (+44) (0)131-248 2901 Project Homepage at http://www.rbge.org.uk/research/Diadist/index.htm RBGE home page at http://www.rbge.org.uk ______________________________
Oxygen monitors are pretty standard in hospital MRI centers. I have some experience with ones from Enmet. URL is http://www.enmet.com/medical.html. Sensor cells need to be replaced at least once a year, and the calibration drifts as the cells age. Although calibration kits are available, I alway just calibrated to the known O2 level in the air, at a place and time when no cryogens were present.
Dave
Shanling Shi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello everyone, } } We have a small room for cryoesctioning and storage of a liquid nitrogen } container. For the safety concern, I would like to get a oxygen sensor or alarm } to monitor the oxygen level. Does anyone konw where I can get this kind of } device? Any suggestion will be appreciated. Thanks in advance. } } Shanling } } Shanling Shi } Advanced Imaging & Measurement } Unilever Research US } 45 River Road } Edgewater, NJ 07020 } 201-840-2340 } Shanling.Shi-at-unilever.com
-- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University voice: 617-627-2163 Fax: 617-627-3443 email: dwilbu01-at-tufts.edu __________________________________
Does anyone have any information or opinion, positive or negative, about Q-Imaging and their Retiga 1300 cooled ccd camera? Their web site is www.qimaging.com. We would be using the camera for fluorescence imaging (with deconvolution) and bright field, dark field, DIC, phase imaging. Please feel free to contact me on or off the list.
Thanks for any info, -- _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ Jessica Wagner Molecular Cytology Core Facility University of Missouri 573-882-4895
Hello all, Does anybody out there know of a decent EM book that focuses on pathology in agricultural systems (i.e. chickens, maize and insects [as vectors])?
Cleaning out my "delete" file I have just noticed a request for a method to enable the cracking of human hair. I have been involved with this type of "problem" many times so set out below is a method which we have found works well for LM as well as SEM. We have even used the technique to investigate failures in freezer bags!
1. Drill two or three holes through two SEM stubs placed face to face. 2. Pass the fibres through the holes. 3. Fix them in place with a water based carbon solution, make sure the fibre/stub interface is well wetted. 4. When fully dry place in liquid nitrogen CARE!!. 5. When the bubbling stops lift out CARE!! 6. Place on an insulating surface and with a blade strike the interface between the two stubs - the system will fracture. 7. When dried off you have two sets of surfaces to look at in LM or SEM.
It works great on a number of differing media other than fibres, the only snag is they must not be affected by the water base.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Direct Line 816512 Fax 814007 www.emcourses.com
I am posting the following for a colleague. Please respond directly to him.
Marie
**********************
Electron Microscopy Technician/Research Assistant
This is a full time position of which 70% of time will be devoted to electron microscopy research support and 30% to a combination of research support in light microscopy immunocytochemistry, cell biology and general laboratory maintenance. Percentages might be adjusted to research demands. Responsibilities include: Transmission electron microscopy (TEM) preparation, immunolabeling and TEM observation and documentation of perfused brain tissues; preparation and immunolabeling of perfused brain tissue samples for light microscopy immunocytochemistry and immunofluorescence; cell culture; preparation of buffer solutions; laboratory maintenance; supervision of laboratory ordering, receiving and record keeping, among others. Qualifications: Background in biological sciences. Candidates with experience in biological transmission electron microscopy and good communication skills are preferred. Salary commensurate with experience.
Submit curriculum vitae plus names, addresses and phone numbers of three references to:
Prof. Angel L. de Blas Dept. Physiology & Neurobiology 3107 Horsebarn Hill Rd; U-4156 Storrs, CT 06269-4156 Ph: (860) 486-3285 Fax: (860) 486-3303 e-mail: deblas-at-oracle.pnb.uconn.edu
We encourage applications from under-represented groups including minorities, women and people with disabilities.
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2242 University of Connecticut Storrs, CT 06269-2242 Phone: 860-486-3588 Fax: 860-486-6369
The following position for a microscopist is being advertised in a major Australian University.
The University of New South Wales is 30 minutes to the central business district of Sydney and 10 minutes to Coogee Beach. The University of NSW is widely regarded as the premier Australian research University and encourages a vigorous research program. The University was the cradle for two major advances in electron microscopy:
1. The Robinson scintillator backscatter detector
2. The Environmental scanning electron microscope
The monetary reward may not seem generous compared with US or European salaries.
But this is compensated for by the strong local purchasing power of the Australian dollar and the benefit that a good lifestyle is much less expensive and more accessible in Sydney than in other international capitals.
Sydney is noted for its mild climate; beaches; opera house; cosmopolitan atmosphere; busy arts scene; access to aquatic sports like swimming, surfing, sailing, SCUBA diving; international standard sports facilities; quality of cuisine and value of wines; excellent educational facilities; high quality of medical care etc. Sydney is an ideal city in which to raise children.
You can learn about NSW from the State Government website
http://www.nsw.gov.au/
You can learn about Sydney from the tourism website
http://www.visitnsw.com.au/0300/0300.asp
You can learn about the University at http://www.unsw.edu.au and about the Electron Microscope Unit at http://srv.emunit.unsw.edu.au
You can direct questions to
p.munroe-at-unsw.edu.au
{bold} {color} {param} ffff,0000,0000 {/param} {bigger} Deputy Director
UNIVERSITY OF NEW SOUTH WALES
ELECTRON MICROSCOPE UNIT
REF. 1293EMAIL
{/bigger} {/color} {/bold}
The Electron Microscope Unit is a central infrastructural research facility, containing eleven principal instruments with a staff of ten. The Unit supports a very wide range of research projects from across the University and its success is based on a strong client-focused orientation. The Unit seeks a dynamic electron microscopist, ideally with a strong background in biomedical or polymer research. The successful applicant will be expected to assist the Unit's Director in the day-to-day management of the Unit, provide expertise in microscopy to a wide range of research projects in biomedical and organic materials and provide appropriate training in microscopy to the Unit's users. The Deputy Director will also develop and carry out research projects, not only related to their own interests, but also in collaboration with other academic staff within the University.
The successful applicant will also be expected to take up a fractional (20%) appointment as Lecturer or Senior Lecturer in a School relevant to their background and experience and will contribute to the teaching effort of that School and to supervise research students as appropriate.
{bold} Additional essential criteria {/bold} for appointment at Senior Lecturer level: a significant record of publication, a demonstrated track record in attracting research funding and prior experience in managing an electron microscope facility.
{bold} Essential criteria: {/bold} a PhD; proven capacity to undertake high quality research; track record of publications in internationally refereed journals; proven experience in teaching; excellent interpersonal, oral and written communication skills; very high level practical experience and theoretical ability in electron microscopy; ability to implement OHS principles and equity and diversity policies and programs.
Desirable criteria: track record in attracting research funding; experience in management of a research laboratory.
{bold} The salary range {/bold} will be commensurate with Lecturer A$52,173 - A$61,957 per year or Senior Lecturer is A$63,912 - A$73,695 per year depending on qualifications and experience.
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Enquiries may be directed to Electron Microscope Unit Director, Associate Professor Paul Munroe, on telephone (61 2) 9385 4435, facsimile (61 2) 9385 6400 or email p.munroe-at-unsw.edu.au.
Applications close 7 February 2002.
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Dr. Mel Dickson,
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How are they mounted? I have no experience with diatoms, but for what it's worth, what about minimizing the refraction BEFORE the light gets to the computer? I mean if the diatoms are in water you may be able to increase the refractive index of the medium you are using (by adding dextran, polyvinyl alcohol, etc) to nearly match that of the specimen, so that the object's refraction is less of a problem.
OptiPrep (TM) (aka iodixanol) is miscible with water and has a very high refractive index, 1.40 for a 40% solution, which is near the R.I. of silica. Another contrast medium, Omnipaque (TM) (iohexol) is lower in R.I. but may be less expensive.
I would guess that this approach would even improve deconvolved images of diatoms in water. Does anyone have experience with this?
Richard
Richard Thrift, PhD
(858) 625-2424 Richard_Thrift-at-SkyePharma.com SkyePharma, Inc 10450 Science Center Drive San Diego, CA, 92121 USA
we are using a light microscope to acquire brightfield images of diatoms, which we then want to process by computer. However, the presence of diffraction effects around the edges of the images is causing us problems.
Does anyone know of any software, preferably free, which can deconvolve an image to reduce the effects of diffraction?
thank you
Micha Bayer ______________________________
Dr. Micha Bayer Royal Botanic Garden Edinburgh 20A Inverleith Row Edinburgh EH3 5LR Scotland, U.K. Tel. (+44) (0)131-248 2915 Fax (+44) (0)131-248 2901 Project Homepage at http://www.rbge.org.uk/research/Diadist/index.htm RBGE home page at http://www.rbge.org.uk ______________________________
With the advent of digital cameras that can be hooked into either eye piece of a stereo microscope, the collection of stereo pairs seems very simple. And with inkjet printers and programs like Adobe Photoshop we now have a great deal of freedom. Has anyone found any simple solutions for printing and presenting the stereo pairs?
I have never worked with diatoms and have never experienced the problem you describe. I have however worked with image processing and analysis. One thing that experience has taught me is that it is best to present as 'good' or 'pure' an image as possible to the computer rather than use the software to 'clean it up'. Such processing will change your image/data in some way which may be undesirable (depends what you are doing).
Could you suspend them in something else? Use a different optical setup?
I would be interested to know if you find an answer.
Good luck.
Nadolig Llawen a Blwddyn Newydd Dda / God Jul och Gott Nytt År / Merry Christmas and a Happy New Year
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Dept of Biomedical Laboratory Science, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sweden
I tried to reply directly to you but it bounced for some reason, so this time I'm sending to the whole list, assuming you'll see it this way. (If anyone else is having problems with a wonky joystick, read on to learn what's fixed mine.)
Frank Thomas GSC Atlantic
----- Original Message ----- } From: "Frank Thomas" {thomasf-at-agc.bio.ns.ca} To: "Monson, Frederick C." {fmonson-at-wcupa.edu} Sent: Tuesday, December 11, 2001 3:07 PM
Hello Micha, I would agree that index matching your mounting media to the refractive index of silica should alleviate the diffraction problems you describe. One important concern is the method by which you are introducing contrast into the specimen for visualization. When the refractive indices are matched, the diatoms will be invisible under standard brightfield illumination. Phase contrast and Nomarski DIC depend on the changes in refractive index at boundries between areas of differing RI to enhance contrast. I have no experience with imaging diatoms, but I seem to recall that darkfield illumination is a popular way to image diatoms, although this method is also dependant on at least a slight mismatch in the refractive index of the specimen and mountant. You might be able to get away with a very close match, however. The numerical aperture of the lens you are using to image the diatoms should be as large as possible. I'm not sure how big your diatoms are, but using a higher numerical aperture objective (preferably oil immersion) may help. If your diatoms are small enough to approach the diffraction-limited resolution limit of the objective lens you are using switch to a higher power or better quality objective (if you have that option). Good luck! -Karl G.
At 12:00 PM 12/11/2001 +0100, Micha Bayer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_______________________________________________ Karl Garsha Specialist in Light Microscopy Beckman Institute for Advanced Science and Technology 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
I just wanted to point you to our analySIS software and the Stereo module. You can take a stereo pair, and display that anaglyphically (red and green), and with the correct glasses you see a 3D image (you can of course also print that). Alternatively, you can have the software calculate the 3D surface of the Object from the stereo pair and then display your object in 3D on the computer. The software was initially written for stereo imaging on an SEM, but there is no reason it should not work on images from a stereo microscope.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Smartech [mailto:smartech-at-optonline.net] Sent: Tuesday, December 11, 2001 9:14 PM To: To all on the list
With the advent of digital cameras that can be hooked into either eye piece of a stereo microscope, the collection of stereo pairs seems very simple. And with inkjet printers and programs like Adobe Photoshop we now have a great deal of freedom. Has anyone found any simple solutions for printing and presenting the stereo pairs?
In a message dated 12/12/01 10:24:00 AM, mb-at-Soft-Imaging.com writes:
} I just wanted to point you to our analySIS software and the Stereo module. } You can take a stereo pair, and display that anaglyphically (red and green), } and with the correct glasses you see a 3D image (you can of course also } print that). Alternatively, you can have the software calculate the 3D } surface of the Object from the stereo pair and then display your object } in 3D on the computer. The software was initially written for stereo imaging } on an SEM, but there is no reason it should not work on images from a stereo } microscope.
Since you originally asked about Photoshop, I'd like to point out that combining two images as an anaglyph is straightforward using the Photoshop channels command (put the left image into the red channel, the right one into the blue and green). And for measuring the parallax and generating a height map or reconstructed 3D image of the surface you can use the Image Processing Tool Kit plug-ins for Photoshop. One of the examples on the web site (see http://www.ReindeerGraphics.com/foveapro2/surface.html) in fact uses images of a small fossil captured through a stereo microscope using an inexpensive Nikon digital camera.
The backstreaming of oil from diffusion pump systems into the specimen chambers of electron microscopes has been a problem that has been around for a long time, and one that is very hard to eliminate. This was a matter that I addressed as fully as I could while writing my book on 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html and http://www.pup.princeton.edu/titles/6484.html for a description). Dealing with backstreaming from rotary vane pumps is described on pages 144 to 150, and from oil diffusion pumps on pages 190 to 200. Perhaps some of the information presented there would be of help. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Hello, I was going to volunteer to give a 60 minute science demonstration to students at local grade schools and was looking for ideas for projects. I came up with some things such as having the students look at organisms in pond water in light microscopes, maybe some SEM photos of common insects (which may not be hands on enough to interest the kids), or teaching them how to pan for gold. The idea is to capture the interest of students and encourage them to study science.
Could you forward any ideas you may have? Thanks, Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Çan anyone direct me to a manufacturer of 2-chamber coverslips for cell culture that use #1.5 thickness coverslips? So far, I'm only finding #1 thickness.
Thanks, Glen
-- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax ************************************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
We're looking for a 100X planapo oil lens (NA = 1.4) for a Nikon Diaphot TMD, tube length 160 mm. If anyone has one in excellent condition to sell, please let me know. Vendor replies welcome. Phil -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
I guarantee that they won't match! Without a bit of directed effort, at least, and if they do happen to match, it is easily fixed by adjusting the concentration of polymer.
With just a tiny bit of mismatch between sample & medium, the sample can be imaged easily by tweaking the contrast on the camera, even if it is not so easily seen by eye.
Another trick which works for latex beads & may work for silica is to add a bit of Nile Red to the suspension. The dye will stick to the surface & fluoresce, while it is almost nonfluorescent in water.
Richard
} } } James Pawley {jbpawley-at-facstaff.wisc.edu} 12/12/01 10:08:29 AM } } } Problem is that when the RI finally does match, bright field contrast disappears.
Jim P. } ------------------------------------------------------------------------ } How are they mounted? I have no experience with diatoms, but for } what it's worth, what about minimizing the refraction BEFORE the } light gets to the computer? I mean if the diatoms are in water you } may be able to increase the refractive index of the medium you are } using (by adding dextran, polyvinyl alcohol, etc) to nearly match } that of the specimen, so that the object's refraction is less of a } problem. } } OptiPrep (TM) (aka iodixanol) is miscible with water and has a very } high refractive index, 1.40 for a 40% solution, which is near the } R.I. of silica. Another contrast medium, Omnipaque (TM) (iohexol) is } lower in R.I. but may be less expensive. } } I would guess that this approach would even improve deconvolved } images of diatoms in water. Does anyone have experience with this? } } Richard } } Richard Thrift, PhD } } (858) 625-2424 Richard_Thrift-at-SkyePharma.com } SkyePharma, Inc } 10450 Science Center Drive } San Diego, CA, 92121 USA } } } } } "Micha Bayer" {M.Bayer-at-rbge.org.uk} 12/11/01 12:00:20 PM } } } } Hi, } } we are using a light microscope to acquire brightfield images of } diatoms, which we then want to process by computer. However, the } presence of diffraction effects around the edges of the images is } causing us problems. } } Does anyone know of any software, preferably free, which can } deconvolve an image to reduce the effects of diffraction? } } thank you } } Micha Bayer } ______________________________ } } Dr. Micha Bayer } Royal Botanic Garden Edinburgh } 20A Inverleith Row } Edinburgh EH3 5LR } Scotland, U.K. } Tel. (+44) (0)131-248 2915 } Fax (+44) (0)131-248 2901 } Project Homepage at http://www.rbge.org.uk/research/Diadist/index.htm } RBGE home page at http://www.rbge.org.uk } ______________________________
-- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Gordon - Try liquid nitrogen experiments. These are always a hit with kids although a but risky so caution is advised. You could freeze and break rubber bands, shatter a rose, inflate a balloon with LN gas boil off (use a plastic soda bottle) then deflate it by placing back into LN. There are lots of other ideas as well but this can get you started. Good luck.
Dave
On Wed, 12 Dec 2001, Gordon Vrololjak wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I was going to volunteer to give a 60 minute science demonstration to } students at local grade schools and was looking for ideas for projects. I } came up with some things such as having the students look at organisms in } pond water in light microscopes, maybe some SEM photos of common insects } (which may not be hands on enough to interest the kids), or teaching them } how to pan for gold. The idea is to capture the interest of students and } encourage them to study science. } } Could you forward any ideas you may have? } Thanks, } Gordon } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } }
I am looking for anyone with experience looking at coccolithophorids using TEM.
We need to look at sections. The guys are only about 5 um in diameter. I have done some thin layer embedding and individual cell selection before, but never on anything this small. I will be checking other sources of information about techniques, but thought I would ask here too,
Any hints or tips on fixing, embedding, etc. that might help?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Dear List, dear Jon, In our lab people from the institute of crystallograhpy investigated coccoliths using TEM. They just used "powders" in plane view. If there is any method to prepare these algae so that the algae or the coccoliths can be sectioned and investigated individually I would be very glad to receive this inforamtion, too. Thank you very much. Gerhard Frank
Jon Krupp schrieb: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Season's Greetings: } } I am looking for anyone with experience looking at coccolithophorids using TEM. } } We need to look at sections. The guys are only about 5 um in diameter. I } have done some thin layer embedding and individual cell selection before, } but never on anything this small. I will be checking other sources of } information about techniques, but thought I would ask here too, } } Any hints or tips on fixing, embedding, etc. that might help? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
-- Gerhard Frank Central Facility for High Resolution Electron Microscopy UNIVERSITAET ERLANGEN-NUERNBERG Institut fuer Werkstoffwissenschaften VII Cauerstr. 6, D-91058 Erlangen, Germany Gerhard.Frank-at-ww.uni.erlangen.de
Caroline Schooley has assembled several microscopy based school activities on the Project Micro web page ( http://www.msa.microscopy.com/ProjectMicro/ClassroomActivities.html). The one I am familiar with, The Beanie Baby Mystery, is easy to do with a few stereomicroscopes or magnifying glasses. It can take 30-45 minutes depending on the size of the class and the story can be changed for the age of the students.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Gordon Vrololjak {gvrdolja-at-nature.Ber To: microscopy-at-sparc5.microscopy.com keley.EDU} cc: Subject: demonstration for K-6th grade students 12/12/01 12:13 PM
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Hello, I was going to volunteer to give a 60 minute science demonstration to students at local grade schools and was looking for ideas for projects. I came up with some things such as having the students look at organisms in pond water in light microscopes, maybe some SEM photos of common insects (which may not be hands on enough to interest the kids), or teaching them how to pan for gold. The idea is to capture the interest of students and encourage them to study science.
Could you forward any ideas you may have? Thanks, Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Nalge Nunc, their Lab Tek II chambered coverglass. They also make 1, 4, and 8 well coverslips with 1.5 thickness coverglasses. I don't have a phone number or URL, as our dealings are with the local distributor. Phil
} Çan anyone direct me to a manufacturer of 2-chamber coverslips for cell } culture that use #1.5 thickness coverslips? So far, I'm only } finding #1 thickness. } } Thanks, } Glen } } -- } Glen MacDonald } Microscopy and Imaging Facility } University of Washington Core for Communication Research } Virginia Merrill Bloedel Hearing Research Center } Box 357923 } University of Washington } Seattle, WA 98195-7923 } glenmac-at-u.washington.edu } (206) 616-4156 (206) 616-1828 fax } ************************************************************************** } C:} The box said "Requires Windows95 or better". So I bought a Macintosh. } **************************************************************************
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
General preparation methods for light microscopy and SEM examination of coccolithophorids are nicely summarized on pages 330-331 of "Plankton Stratigraphy" Bolli, Saunders and Perch-Neilsen (Editors), 1985, Cambridge University Press. They don't, however, say anything about sectioning them. For biostratigraphy, where it's important to know exactly what species you're dealing with, you normally want the little guys as whole as possible, and id's are made on the basis of (mostly) morphology. I bet your local Earth Sciences library has this book, anyway. Are you at liberty to say why you're interested in sectioning them?
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, December 12, 2001 6:20 PM
Calling All Microscopists,
We have often removed sputtered gold from SEM specimens with a solution of potassium iodide / iodine complex. However, I've forgotten the quantitative formulation. Does anyone know how much of KI and of I2 to use?
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
I would like to direct you to a wonderful program called "Project MICRO" (details can be found on the MSA website): http://www.msa.microscopy.com/Project Micro
Basically it is a program to introduce microscopic explorations to elementary and junior high school students. I am a member and corresponding secretary of MSA's local affiliate, New England Society for Microscopy, and our group has wholeheartedly embraced Project MICRO. We actually raised money and put together three kits (inlcudes all the materials, dissecting and simple monocular microscopes). Several members have used the program in their local schools. We even had a workshop at our annual Woods Hole Meeting (Woods Hole, MA) a few years back to encourage members to get involved. I am hoping to run a Saturday Workshop at a local school sometime in January.
The program utilizes a workbook "Microscopic Explorations"-a GEMS Festival Teacher's Guide, which was put together at the Lawrence Hall of Science at the University of California at Berkeley.
It sounds like a good program for you. You could get a kit and run a workshop with teachers in a local school. Students and teachers are overwhelmingly supportive.
Take a look......
Peggy Sherwood
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-- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
At 08:34 AM 12/13/2001 -0600, you wrote: } Microscopy-at-sparc5.microscopy.com
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
I am currently working on an EDS analysis project on carbon / fluorine matrices. Usually carbon starts as the taller peak, and remains so throughout the acquisition. Occasionally, however, when starting an spectrum acquisition I will see the fluorine peak start as the taller peak, then slow to become the shorter peak.
I was told by an observer that there is a phenomenon known as "chemical shift," that could have an impact on the amount of certain elements in the interaction volume of a sample. This person didn't know much about it other than it could give you inaccurate quant results.
Is there such a thing as chemical shift, and if so, could someone explain what exactly is happening during the process?
This sounds like a problem with terminology. I would not call it chemical shift but probably would call it devolatilization.
When someone says "chemical shift", I usually think of a shift in the energy or wavelength of the x-ray lines due to the chemical environment of an atom. For example, sulfur as sulfide, as sulfate, and as thiol each have slightly different peak energies. That is something that can be observed and can be a problem with wavelength dispersive analysis. However, the resolution is not so good to be able to observe such a shift, unless you have one of the new microcalorimeter detectors.
I might also consider a shift of intensity between peaks as a chemical shift. I remember seeing a paper about the L-lines of nickel. Depending on the amount of alloying element (Al, I think), the relative intensity of the L-lines changed dramatically. The explanation was that the electronic environment changed so that different transitions giving rise to L-lines were more favorable as there was more or less of the alloying element. This is quite visible by EDS, but not that common.
I sincerely doubt that you are seeing either of these effects.
What you are probably seeing is a change in chemical composition with continued exposure to the electron beam. That does lead to a shift in the chemical composition, but I reserve the phrase chemical shift for what I explained above. You are either preferentially volatilizing one component compared to another, or you are changing the components through heating. This is not uncommon, particularly at higher currents and voltages.
You might try a number of things to reduce the energy dumped into your sample. You might try dropping your beam current. If you are doing SEM, you might also reduce the voltage. You might try spreading a beam over an area (or larger area) rather than using a point analysis so that the energy is spread out. (This presumes your sample is homogeneous.)
You might also try a much shorter exposure, or successive exposures to try to capture the initial composition. I know our Tracor Northern TN-2000 had a feature for painting a window for an element and plotting the intensity of that window over time - you picked the time interval per channel. However, I don't think that feature has been on any of our last three EDS systems. It would be really helpful for what you are facing. There are probably ways to fake your way through with what you do have. \
I hope this helps. Warren
At 12:24 PM 12/13/01 -0600, you wrote:
} I am currently working on an EDS analysis project on carbon / fluorine } matrices. Usually carbon starts as the taller peak, and remains so } throughout the acquisition. Occasionally, however, when starting an } spectrum acquisition I will see the fluorine peak start as the taller peak, } then slow to become the shorter peak. } } I was told by an observer that there is a phenomenon known as "chemical } shift," that could have an impact on the amount of certain elements in the } interaction volume of a sample. This person didn't know much about it other } than it could give you inaccurate quant results. } } Is there such a thing as chemical shift, and if so, could someone explain } what exactly is happening during the process? } } Thanks, } } } Jeff Oakley } Rayovac Corporation
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I am looking for help finding staining chemistries or procedures, which may be used to enhance imaging of GaN based material layers. Any information you could pass along would be helpful.
Robert H.Olley wrote: =============================================================
We have often removed sputtered gold from SEM specimens with a solution of potassium iodide / iodine complex. However, I've forgotten the quantitative formulation. Does anyone know how much of KI and of I2 to use? =============================================================== I know it goes against conventional wisdom, but in the past we have been able to remove the sputtered gold layer using an oxygen plasma in one of our Plasma Prep II plasma etchers. I have never had anyone explain to me why it would do this but it apparently does. A typical gold thickness would come off in about thirty or so minutes. We made this discovery when trying to come up with a good demonstration of the effect of etching as a function of time on some ordinary clay coated paper. To etch a little more we had to first etch off the applied gold layer for the previous step. This was done some years ago before the days or the more modern techniques that often times require no coating at all. We have the results from that study on a sign we use in our exhibit stands but don't have it yet on our website. If you would like to see the results, ask me when ever you see SPI at a trade show exhibition. I usually have it with me on display.
For some samples, the liquid method would change the sample and that would certainly be the case for paper.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
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It's also possible that carbon is increasing during the analysis due to hydrocarbon contamination of the sample or microscope, a common situation in e-beam analysis. You might try monitoring the count rates for the C and F x-rays to see whether the carbon is increasing or fluorine is being lost.
Larry Thomas Pacific Northwest National Laboratory P.O. Box 999 Mail Stop P8-16 Richland, WA, USA
---------- From: Warren E Straszheim Sent: Thursday, December 13, 2001 12:11 PM To: Oakley, Jeff Cc: Microscopy-at-sparc5.microscopy.com Subject: Re: EDS analysis / "Chemical shift"
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This sounds like a problem with terminology. I would not call it chemical shift but probably would call it devolatilization.
When someone says "chemical shift", I usually think of a shift in the energy or wavelength of the x-ray lines due to the chemical environment of an atom. For example, sulfur as sulfide, as sulfate, and as thiol each have slightly different peak energies. That is something that can be observed and can be a problem with wavelength dispersive analysis. However, the resolution is not so good to be able to observe such a shift, unless you have one of the new microcalorimeter detectors.
I might also consider a shift of intensity between peaks as a chemical shift. I remember seeing a paper about the L-lines of nickel. Depending on the amount of alloying element (Al, I think), the relative intensity of the L-lines changed dramatically. The explanation was that the electronic environment changed so that different transitions giving rise to L-lines were more favorable as there was more or less of the alloying element. This is quite visible by EDS, but not that common.
I sincerely doubt that you are seeing either of these effects.
What you are probably seeing is a change in chemical composition with continued exposure to the electron beam. That does lead to a shift in the chemical composition, but I reserve the phrase chemical shift for what I explained above. You are either preferentially volatilizing one component compared to another, or you are changing the components through heating. This is not uncommon, particularly at higher currents and voltages.
You might try a number of things to reduce the energy dumped into your sample. You might try dropping your beam current. If you are doing SEM, you might also reduce the voltage. You might try spreading a beam over an area (or larger area) rather than using a point analysis so that the energy is spread out. (This presumes your sample is homogeneous.)
You might also try a much shorter exposure, or successive exposures to try to capture the initial composition. I know our Tracor Northern TN-2000 had a feature for painting a window for an element and plotting the intensity of that window over time - you picked the time interval per channel.
However, I don't think that feature has been on any of our last three EDS systems. It would be really helpful for what you are facing. There are probably ways to fake your way through with what you do have. \
I hope this helps. Warren
At 12:24 PM 12/13/01 -0600, you wrote:
} I am currently working on an EDS analysis project on carbon / fluorine } matrices. Usually carbon starts as the taller peak, and remains so } throughout the acquisition. Occasionally, however, when starting an } spectrum acquisition I will see the fluorine peak start as the taller peak, } then slow to become the shorter peak. } } I was told by an observer that there is a phenomenon known as "chemical } shift," that could have an impact on the amount of certain elements in the } interaction volume of a sample. This person didn't know much about it other } than it could give you inaccurate quant results. } } Is there such a thing as chemical shift, and if so, could someone explain } what exactly is happening during the process? } } Thanks, } } } Jeff Oakley } Rayovac Corporation
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Does anyone know of an antifade agent that works for liver-cell microscopy systems? We're having trouble with fading of the fluorescent signal. Thanks in advance.
Mary
Mary McKee MGH Renal Unit Charlestown, MA 02129 (617)726-3696
I am working ICs which have an upper and lower TiW barrier metal layer between AlSi. The problem is that the huge extreme of Z makes SE and BSE imaging big time difficult. I tried moderating the range using Au/Pd (60/40%) but it makes little difference. This coating was about 40A thick.
Does anyone have a suggestion about how to normalize the Z range of materials in this type of specimen so that either SE or BSE images would show reasonable contrast? If anyone would like to see my pathetic images at this time, I can put them on one of my web sites.
Here are the killer (Z values): Al=13 Si=14 Ti=22 W=74 (low % but huge impact on image contrast)
I have thought about Silver (Z47) and Zirconium (Z40) to try to even out the overall Z range of the specimen. Unfortunately, Anatech does not make Zirconium targets for the Hummer VII. I have available Au, Pd, Pt, silver, or air.
Am I at the limits of this coater and its target materials?
Our ion mill has developed a vacuum problem. After checking and servicing various components including replacing the whisperlock with the blanking tool, we have narrowed down the possible defect to be with the vac valve (airlock evacuation valve) on the right mill. The vacuum deteriorates significantly when the right mill vac button is depressed and remains at the poor vac continuously. This is irrespective of whether the whisperlock or the blanking tool is installed . Among the spares, we have spare o-rings for the vac and vent valves, but we do not have any information regarding how these are to be greased / replaced or for the valve to be serviced. No apparent screws / nuts / washers etc are found. Any information regarding the servicing of these parts will be greatly appreciated since we do not have a service contract on the instrument. Our instrument is nearly 10 years old and we have been maintaining it ourselves for most part of the period.
With Thanks and Best wishes to all list members, ---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
I wish you had said more about the materials you are looking at. The electron beam causes considerable heating in a sample, and if there is free fluorine in the sample, not chemically bonded, then the action of this heating and the vacuum could cause migration of the fluorine. As mentioned by another poster, 'chemical shift' in x-ray spectroscopy refers to the actual shifts in peak locations caused by chemical bonds altering the quantum energy levels of the electron shells. This effect will only be seen on fully focusing Johannson wavelength spectrometers or the more recent microcalorimeter systems, developed at NIST.
One other possibility is an instrumental problem. Problems in the DAC circuitry of an EDS can result in odd effects, particularily at the low end. Normally, something like this would be heat related and probably show up as the instrument is first warming up. Is it possible that the high fluorine peaks that slowly decrease are found primarily on the first spe ctrum or two in a run of samples?
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
-----Original Message----- } From: Oakley, Jeff [SMTP:oakleyj-at-rayovac.com] Sent: Thursday, December 13, 2001 10:25 AM To: 'Microscopy-at-MSA.Microscopy.Com'
I am currently working on an EDS analysis project on carbon / fluorine matrices. Usually carbon starts as the taller peak, and remains so throughout the acquisition. Occasionally, however, when starting an spectrum acquisition I will see the fluorine peak start as the taller peak, then slow to become the shorter peak.
I was told by an observer that there is a phenomenon known as "chemical shift," that could have an impact on the amount of certain elements in the interaction volume of a sample. This person didn't know much about it other than it could give you inaccurate quant results.
Is there such a thing as chemical shift, and if so, could someone explain what exactly is happening during the process?
Gary, You should have a gamma function that will allow non-linear amplification on your AMRAY. Have you tried that?
Ken Converse owner Quality Images third party SEM service Delta, PA
Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } hello listers: } } I am working ICs which have an upper and lower } TiW barrier metal layer between AlSi. The problem is } that the huge extreme of Z makes SE and BSE imaging } big time difficult. I tried moderating the range using } Au/Pd (60/40%) but it makes little difference. This } coating was about 40A thick. } } Does anyone have a suggestion about how to normalize } the Z range of materials in this type of specimen so that } either SE or BSE images would show reasonable contrast? } If anyone would like to see my pathetic images at this time, } I can put them on one of my web sites. } } Here are the killer (Z values): } Al=13 } Si=14 } Ti=22 } W=74 (low % but huge impact on image contrast) } } I have thought about Silver (Z47) and Zirconium (Z40) } to try to even out the overall Z range of the specimen. } Unfortunately, Anatech does not make Zirconium } targets for the Hummer VII. I have available Au, Pd, Pt, silver, or } air. } } Am I at the limits of this coater and its target materials? } } TIA, } gary g. } } } }
Sounds like gamma filtering would help you. If you have that option on your SEM, give it a try.
Diane Ciaburi
Gary Gaugler {gary-at-gaugler.com} 12/14/01 12:10 AM
To: MSA listserver {Microscopy-at-sparc5.microscopy.com} cc: Subject: IC cross sections -- with high Z contrast
hello listers:
I am working ICs which have an upper and lower TiW barrier metal layer between AlSi. The problem is that the huge extreme of Z makes SE and BSE imaging big time difficult. I tried moderating the range using Au/Pd (60/40%) but it makes little difference. This coating was about 40A thick.
Does anyone have a suggestion about how to normalize the Z range of materials in this type of specimen so that either SE or BSE images would show reasonable contrast? If anyone would like to see my pathetic images at this time, I can put them on one of my web sites.
Here are the killer (Z values): Al=13 Si=14 Ti=22 W=74 (low % but huge impact on image contrast)
I have thought about Silver (Z47) and Zirconium (Z40) to try to even out the overall Z range of the specimen. Unfortunately, Anatech does not make Zirconium targets for the Hummer VII. I have available Au, Pd, Pt, silver, or air.
Am I at the limits of this coater and its target materials?
Unfortunately, application of a coating will not solve your problem. The only effect will be to reduce sensitivity to changes in Z under the coating. The same effect could be had by just reducing the gain/contrast control. The Z ratio remains the same under the coating. If the contrast/gain is increased to compensate for the sensitivity loss generated by an applied coating, you will be back where you started, but with a degraded signal to noise ratio. I have graphics of this effect in my homegrown "SEM Primer" booklet.
The problem is common for me. Sometimes I want to see very small Z changes when the overall range is huge. You ought to try heterogeneous mixes/compounds like C, O, Al, and U!!!
To my knowledge, there is no really *good* solution to maintaining high BSE sensitivity while not saturating, but I have used two methods.
1) Multiple images: 2, maybe 3 images, each covering a band of atomic numbers without black/white saturation. The images may be used separately, or colored and an overlay composite made using an image editor.
2) Use non-linear video processing. I modified my (previous GW Elec.) BSE system by adding a clipping diode with a variable reference bias . Could set white clipping just about anywhere in the output range. The clipping/limiting was not "hard" so that some signal variation would make it through. This had the effect of reducing effective gain/brightness on the high end. The degree of gain compression was a function of the original gain & brightness and the new adjustment.
Although only viewable effectively using a software package, another approach would to use more than 256 shades/8 bit gray scale (hardware/software permitting). For Example, a 32 bit gray scale image would likely work fine, but you could only "see" a portion of the range at any time.
Woody White McDermott Technology, Inc.
} -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Friday, December 14, 2001 12:10 AM } To: MSA listserver } Subject: IC cross sections -- with high Z contrast } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } hello listers: } } I am working ICs which have an upper and lower } TiW barrier metal layer between AlSi. The problem is } that the huge extreme of Z makes SE and BSE imaging } big time difficult. I tried moderating the range using } Au/Pd (60/40%) but it makes little difference. This } coating was about 40A thick. } } Does anyone have a suggestion about how to normalize } the Z range of materials in this type of specimen so that } either SE or BSE images would show reasonable contrast? } If anyone would like to see my pathetic images at this time, } I can put them on one of my web sites. } } Here are the killer (Z values): } Al=13 } Si=14 } Ti=22 } W=74 (low % but huge impact on image contrast) } } I have thought about Silver (Z47) and Zirconium (Z40) } to try to even out the overall Z range of the specimen. } Unfortunately, Anatech does not make Zirconium } targets for the Hummer VII. I have available Au, Pd, Pt, silver, or } air. } } Am I at the limits of this coater and its target materials? } } TIA, } gary g. } }
I second all the comments made by Warren Straszheim in reply to this posting, but would also suggest what seems to me to be an equally likely possibility given the elements in question - contamination.
Jeff does not mention the form of the samples or the conditions of the experiment, but the observation that the carbon signal increases with analysis time is very common, and is the result of the build-up of contamination resulting from the break-down of hydrocarbon molecules diffusing around the sample surface (sometimes originating from the microscope vacuum, sometime3s carried in with the sample, often both). This is observed much more at high magnifications or in point mode, and both in the SEM and the STEM/TEM. Frequently, a careful examination of an SEM image taken after the analysis will show a spot where the probe had been (in the STEM/TEM the contamination is usually very obvious).
It can be very difficult to predict when you will see contamination problems, and when you won't. Even two samples prepared, supposedly, in the same way, and examined one after the other, may behave differently.
Tony G-R
At 12:24 PM 12/13/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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I am considering purchasing a refurbished fluorescence microscope from Microgen Optics. Any feedback re: experience with this company would be much appreciated. Thanks,
Susan Stanton, Ph.D. College of Allied Health Sciences University of Cincinnati Cincinnati, OH 45267-0394
Chemical shift (changing of the position of the peak due to changing energy of the outer electrons because of the chemical interaction) for fluorine should be relatively small. It usually important only for WDS acquisition if it is performed on standard position. For EDS it should be irrelevant since the whole spectra is acquired and chemical shift will affect mostly position and not intensity of a peak.
I think you run into one of the two (or both) problems: 1. Beam induced migration of fluorine (most probable). 2. Carbon contamination under the beam.
You can try to apply methods suitable for beam sensitive specimens. - Use only low magnifications for very fast observing of the specimen. - Make all preparations at higher magnifications - focusing and whatever else - on one place, then move to desired place and perform acquisition as quickly as possible. - Defocus beam as much as possible. - Use shorter acquisition times and lower voltage and beam current.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Oakley, Jeff [mailto:oakleyj-at-rayovac.com] } Sent: Thursday, December 13, 2001 12:25 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: EDS analysis / "Chemical shift" } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } I am currently working on an EDS analysis project on carbon / fluorine } matrices. Usually carbon starts as the taller peak, and remains so } throughout the acquisition. Occasionally, however, when starting an } spectrum acquisition I will see the fluorine peak start as } the taller peak, } then slow to become the shorter peak. } } I was told by an observer that there is a phenomenon known as } "chemical } shift," that could have an impact on the amount of certain } elements in the } interaction volume of a sample. This person didn't know much } about it other } than it could give you inaccurate quant results. } } Is there such a thing as chemical shift, and if so, could } someone explain } what exactly is happening during the process? } } Thanks, } } } Jeff Oakley } Rayovac Corporation } } }
Lots of good suggestions. Several questions I did not answer. First, the SEM is an AMRAY 1910 Schottky FESEM. Over a reasonable condenser setting range, the probe diameter is about 90A at high condenser setting, 10KV. I have a new Keithley picoammeter on order to use to re-do the probe diameter measurements and record specimen currents. My DMM approach does/did not work.
The cross section sandwich is non-conducting by its nature. Since the device is a completed IC, it has all underlayers, poly, interlayer dielectric, double metal interconnect, 2KA SiO2 and 10KA SiN passivations. Top down, this device would charge. I can image it at moderate KV and small probe diameter with BSE.
My BSE is a Robinson Model 6. I can mix any amount of SE and BSE in any relationship. SE+BSE, SE-BSE, BSE-SE, etc.
My scope has a Gamma control. However, it does not work. This could be because I'm not using it correctly or have not even turned it on. I will check this for sure. If it is broken, Amray will fix it. Gamma is a good idea to try.
The other idea of separate images of different Z areas of interest has potential. How different this would be from the mixer on the Robinson remains to be seen. Even so, separate images could be shot and aligned afterwards.
Procedurally, the cross sections are made with traditional Buehler equipment and fine diamond paste polish. Specimens without barrier metal turn out quite well. This new lot with TiW is a real problem. The next one has TiN barrier metal. The TiN devices are built on thin SOS. The ones I'm working with now are bulk CMOS. The TiW devices will shift to SONOS early next year.
The consensus seems to be that coating is a waste of time. I can't really argue with that, based on my bad prior history of doing this. I have a new specimen with TiW which is mounted and desiccated overnight. I will try looking at it without any coating. The main glass slide is stuck down on a 90 degree Pella stub and the bottom portion is covered with coloidal silver. The upper part of the specimen is un-touched.
Thanks to all. gary g.
At 03:59 AM 12/14/2001, you wrote: } You didn't mention what type of SEM you are using. I assume that you are } looking at cross-sections of the IC's. We use a FESEM and use 2.5kv and } that works nicely without any coating at all. Before we had the FESEM we } used a gold coater. We did not coat the cross-section directly but laid it } flat down so the gold just drifted over the edge of the cross-section. This } gave us very nice contrast with the metalization and the SiO2. Let me know } how this works for you or give me a call. } Mark } 612-954-2845
Thanks to all for the many excellent suggestions. Some ideas had already been tried while others had not.
Gamma is a good idea. My scope has gamma feature but for some reason, it does not work. A service call should solve that problem. But I found a very nice way to handle the huge Z contrast range in barrier metal ICs.
I still have to coat the specimen with only about 30A of Au/Pd or Pt to eliminate charging. I shoot at between 5-10KV with medium spot size. At this range, the key is to do a SE-BSE image with the amount of subtracted BSE set for best image. This seems rather obvious right now. But it did not this AM.
Setting up the SE for excellent detail and definition in the low Z areas while ignoring the high Z areas is the first action. Then, engage mixing, set for SE-BSE and adjust the amount of SE to BSE so that the overall contrast range is evened out. The results are perfect, I think.
you can see a sample at:
http://photoweb.net/Test7-5dsp8bc.jpg
If anyone has any other ideas or suggestions, please don't hesitate to put them forward.
Thanks, Gary Gaugler, Ph.D. Microtechnics, Inc. Granite Bay, CA 916.791.8191 916.791.8186 fax
Start off the New Year with plans to attend the annual MSA/MAs meeting in Quebec August 4-8 2002. This year, the Education Outreach symposium will be examining the use of image processing as a tool in the academic curriculum. In addition to using image processing as a way to increase the use of microscopic images in classes, Ken and I are interested in new approaches for microscopy or remote access to expand the role of microscopy and microscopes in the classroom at any level.
If you are interested in presenting in this symposium, please contact me. I have included the symposium abstract for your information.
Please note: we are more interested in discussing image processing as a curriculum tool than in discussing 'how to' process images.
See you all in Quebec Steve Barlow
Teaching and Learning - Creating Effective, Innovative Solutions in Microscopy, Imaging, and Analysis Steve Barlow and Ken Baker
Teaching and learning in the field of microscopy imaging and analysis are important to the practice of all forms of microscopy. Microscopy imaging and analysis has adopted an extensive collection of tools and techniques and continues to develop as an increasingly complex subject area. Computer-assisted microscopies, high speed global internet communications, remote microscopies and image acquisition, digital imaging tools, and multidimensional imaging and analysis techniques all offer opportunities for powerful innovation in teaching and learning. Pedagogical resources include: distance learning, on-line manuals, references, tutorials, image databases, and a selection of shared utilities and dedicated discussion forums. This symposium will encourage contributions that emphasize creative, effective, and innovative educational approaches to microscopy, image acquisition and analysis, and increased use of microscopy and images in a student curriculum.
dear colleagues I would like to add anatomy of the beetle. I am looking for nice scan picture of the beetle both sides...
Thank you very much for your help.
Marry Christmas
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
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on 12/14/01 6:04 AM, Allen Sampson at ars-at-sem.com wrote:
} } As mentioned } by another poster, 'chemical shift' in x-ray spectroscopy refers to the } actual shifts in peak locations caused by chemical bonds altering the } quantum energy levels of the electron shells. This effect will only be } seen on fully focusing Johannson wavelength spectrometers or the more } recent microcalorimeter systems, developed at NIST. } In the interest of completeness, the tunnel junction diode detector, also recently developed, has energy resolution in the few eV range, so it is also able to detect chemical shifts. Yours, Bill Tivol
I have no experience with TEM investigation of coccolithophorids, but for SEM, the folks here spray them onto stubs - they're so tiny they adhere on their own - and then sputter coat. Also, filtering seawater with coccos in it and looking at a cutout from the filter paper (in the SEM, at least) works well for us.
Good luck, Dee }
} Season's Greetings: } } I am looking for anyone with experience looking at coccolithophorids using } TEM. } } We need to look at sections. The guys are only about 5 um in diameter. I } have done some thin layer embedding and individual cell selection before, } but never on anything this small. I will be checking other sources of } information about techniques, but thought I would ask here too, } } Any hints or tips on fixing, embedding, etc. that might help? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
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Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
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Does anyone know of an antifade agent that works for liver-cell microscopy systems? We're having trouble with fading of the fluorescent signal. Thanks in advance.
Mary
Mary McKee MGH Renal Unit Charlestown, MA 02129 (617)726-3696
I am posting a summary of responses to my request to this list on 3 December 2001, for information about any roles microscopic artifacts may have played in the history of biology. I posted my own experience regarding Sidney Hickson's description of reproductive events in _Millepora_ sp)p). (A copy of my somewhat lengthy post is enclosed at the end of this summary.)
To be fair to Sidney Hickson, who drew and described the incredible, imagined sequence of reproductive events in Millepora sp(p). I mentioned in my original , I believe that he did print a retraction of the findings in that paper.
Gareth Morgan (whose contribution is mentioned below) offered the following remark:
We tend to see what we expect might possibly be there and consciously or unconsciously ignore the rest as 'noise' - perhaps a natural action but also potentially dangerous.
Maureen A. Peterson offered a similar remark, which I took as personal advice:
"Lesson? I better keep my eyes and my mind open."
Both Volker Brinkmann and Mike Dalbey called attention to the "homunculus" (little man-shaped pre-embryos) which were reported in the 18th Century in human sperm.
Volker Brinkmann:
The first microscopic misinterpretation was probably that by Antoni van Leeuwenhook analysing human sperm. He was convinced to see little men with heads, arms and legs swimming around. There are some nice drawings of that.
Mike Dalbey:
Surely the most famous misinterpretations of microscopical observations are those recorded in drawings made by preformationists in the 18th Century of the "homunculus" in the head of human sperm. Several of these drawings are commonly reprinted in textbooks as cautionary examples.
Mike Dalbey further pointed out several interesting examples, including a contemporary issue, the putative bacteria fossiles in the martian meteorite::
For a discussion of a common histological artifact see "Multinucleate Plant Cells" by Burkholder and Mc Veigh (1941) in vol. 68 of the Bulletin of the Torrey Botanical Club p. 395.
You should also check out what the web has to offer on the history of Royal Rife and the Rife Microscope.
Finally, evidence seems to be accumulating that the "nanobacteria" observed by EM in the martian meteorite are artifacts.
Maurreen A. Peterson (Mape-at-mail.ifas.ufl.edu) posted concerning a disease of grapes, a case of failure to see the etiological agent, which was there all the time:
Being a Plant Pathologist, the best I can think of is the story of Pierce's disease of grapes, which was of unknown etiology for a long time. Forgive me for not seeking details from the literature.
At one time it was proposed to be caused by a virus. Extensive light microscope work missed the true cause- a bacterium. After elucidation of the pathogen, I am told that upon review of work previously done, the bacterium WAS there to be seen, but was missed.
Upon a similar vein, MLO's in plants were not recognized in TEM work until a human or animal pathologist saw micrographs of MLO's in plant tissue, and was readily able to say what they were.
Anecdotal, but food for thought.
Two similar misinterpretations of existing evidence were pointed out by Gareth Morgan ( {Gareth.Morgan-at-impi.ki.se} ). The first regarded _Helicobacter pylori_, a causitive agent for gastric ulcers.
Not sure if this one fits the bill but it is a similar one to the grape disease story by Maureen Petersen.
The one I mean is the discovery/description of Helicobacter pylori in human gastric biopsies in the early 80's (when it was given the name Campylobacter pylori). It had been there all of the time but had been disregarded as 'stuff'.
Gareth suggested a google search might reveal more information, and indeed it did: over 82,000 hits. I hope I will be forgiven for quoting one site in part:
Although they were the first to succeed in establishing a link between bacteria and ulcers, the Australians were not the first to try. Since the time of Robert Koch in the late19th century, microscopists observed curved bacteria among the cells under the mucus lining of the stomach, particularly in and around ulcer craters. No one had ever succeeded in isolating the microorganisms, however, and their presence had been explained away as artifact or postmortem contamination. Besides, it was thoroughly accepted among clinical microbiologists of the time that it was unlikely that bacteria could live and grow in the strongly acidic environment of the human stomach.
Warren, a pathologist who examined gastric biopsies, also observed the curved rod-shaped bacteria under his microscope. After examining many such specimens, he realized that the bacteria were always present in tissue that showed signs of inflammation, that the number of organisms correlated with the degree of the inflammation present, and that they occurred in half of the routine gastric biopsy specimens he examined. Convinced that his observations were significant and merited further investigation, he kindled the interest of Barry Marshall, then a trainee in internal medicine, and together they set out to isolate the source of the infection.
[This site is: http://www.faseb.org/opar/pylori/pylori.html]
Gareth also suggested a similar misreading may have been involved with "the finding of Actinomyces in cervical smear material." I was unable to find any clear elucidation of such events; however, one site mentioned that care should be taken by the pathologist in diagnosing "atypical _Mycobacteria_" due to possible presence of Actinomycetes.
H. F. Moura Nunes posted about another possible case:
I am not sure of that. But I heard some years ago that the first information about the structure of poliovirus - in the early days of the electron microscope - was that they were long filamentous structures. Some time later it has been seen that the filamentous structures were in fact bacterial flagella.
Tina Carvalho posted a long discussion of a personal experience regarding another real structure that had been interpreted as an artifact. The experience must be re-posted in its entirety, in spite of its length.
Although I 'm sure there are examples of misinterpretation of artifacts that have been perpetuated for many years, I can offer here a story of the flip side. This is about what appeared to be a well-known artifact that turned out to be, in fact, a surprising real structure. It's also an embarrassing personal story!
An undergraduate student was working with some colleagues who were studying the behavior and neurophysiology of the escape response of several species of planktonic copepods (small crustaceans). They had reams of data and were beginning to characterize the responses of the animals to different kinds of stimuli. Each species had it's own characteristic response, but a general picture was beginning to form. There appeared to be two different classes of response though, one type particularly puzzling. The undergrad spent some time in the EM facility with me, using SEM to describe the sensory setae. She expressed an interest in trying TEM and so we went through fixation, embedding, sectioning, etc. I had worked on the tiny beasts for close to 10 years and had found some interesting structures that I worked on from time to time. Copepods were difficult to fix, some species more so than others. So I had concentrated on the "pretty" ones and temporarily given up on the "ugly" ones - the ones with all those myelin body artifacts that I just couldn't seem to avoid. Finally it was time to try to work on them again, so I gave this one particularly difficult species to the undergrad to try to fix and section. She came to me with micrographs, wondering what in the world were those squiggles she saw. I patiently explained that myelin bodies were a fixation artifact, produced when lipids became mobile during fixation, then reformed in these "onion bodies" . I sent her back to work to try again, and she came up with the same results. "What if they aren't an artifact?" she kept asking. I explained that invertebrates do not have such membranous structures; they were artifactual. I brought out the books and papers about myelin body artifacts. I brought out the books and papers that said invertebrates do not have membranous wrappings around their axons. She was unconvinced. In a hurry one day when she was really bugging me I dug through my files and found the decade-old folder to show her I'd seen the same thing - an artifact. "But there's an axon in the middle of each one", she protested, not knowing that was "impossible". I glanced at them impatiently and saw that she was right, but she was running off to class.
Later that day we looked closely and decided that, indeed, the images were a real mess and difficult to interpret, but there was a faint possibility that these weren't classic myelin body artifacts. Whatever it was was certainly reproducible, and these forms appeared every time in the antennal nerve. So we emailed the PIs of the project and told them we might have myelinated axons in the difficult-to-interpret species. It was April1, April Fools' Day, the undergrad's name was April, and they thought it was a joke. After all, the dogma is that invertebrates don't have myelin! Check any biology book.
But by the next day they realized that myelin (OK, "myelin-like structures" for you purists) would instantly and completely explain 10 years' worth of puzzling data.
It took awhile to prove to ourselves (mainly me, since I'd been repeating the dogma for many years) that these structures were real, orderly, and extensive wrappings of membrane around the axons. It took ultrarapid cryofixation and cryosubstitution to really show that the artifactual bodies that had plagued a number of people working on these critters were in truth these surprising, dogma-busting structures.
We made the April the first author on the Nature paper, which came out in April 1999.
I don't take anything for granted anymore! The lesson is: keep an open mind.
For more on the copepod story- Copepod neuroecology http://www.pbrc.hawaii.edu/~petra/copepod.html
Finally, perhaps appropriately, I include my original posting:
I am interested in gaining insight into the role of microscopic artifacts in the history of biology. May I impose on list members to contribute particularly glaring examples of misinterpreation of biological facts due to improper microscopic technique? I apologize if this is off-topic or a waste of bandwidth.
Let me provide the first example of a misinterpretation and request your assistance in learning whether this was due to improper use of the microscope, or perhaps even malfeasance: Sidney Hickson's early work on Millepora spp. (Cnidaria:Hydrozoa) fire corals. It seems an incredible lapse, a wholly fabricated natural history account, one that persisted for a considerable period in the fabric of the mythology of biological knowledge. I am interested because reproduction of Millepora platyphylla is the subject of incompleted thesis research of mine.
Hickson published a report on reproduction of "Millepora" around the end of the 19th Century. (Anong his other errors he synonomyized all species of Millepora as ecomorphs of one, M. alcicornis.) In this report, which I do not have available at this time, he included several plates of drawings depicting a putative sequence of reproductive events in this organism. We now understand that his depiction is not even close to the way that Millepora spp. (which were later redesignated as proper individual species through painstaking work by Boschma---notwithstanding the issues recently raised by molecular work) reproduce. The depiction involved dozens of drawings, and a sequence of events based on a misinterpretation of what are apparently artifacts.
Hickson (of Cambridge University) worked extensively in the field, including Indonesia and the Philippines. Was his microscopic work done in the field? Are members of this list enlightened as to Hickson's methods?
Hickson's erroneous drawings of the medusae of Millepora lived on for over 3/4 of a century in virtually every Invertebrate Zoology textbook published until the late 1980s or 1990s. His erroneous description of the medusa of Millepora as lacking a velum led to the designation of a separate branch of hydromedusae by Mayer, as the only hydrozoan medusa without a velum. My unpublished observations in the 1980s as well as published observations by John Lewis of McGill University showed that the medusae of Millepora spp. clearly possess a velum.
I apologize for monopolizing the bandwidth. I hope this is as fascinating a topic for others as for myself, and not considered off-topic.
-- Alan E. Davis, Science Instructor Marianas High School PMB 30, Box 10006, Saipan, MP 96950 Northern Mariana Islands adavis-at-saipan.com
"An inviscid theory of flow renders the screw useless, but the need for one non-existent." ---Lord Raleigh(aka John William Strutt),or else his son, Jr., who was also a scientist.
I'd be willing to work on them at $75 plus parts. Most of the problems are cleaning of the stick and guides. Other problems are dirty pots.
Send 'em to me and I can fix them and send them back.
No guarantee, but its worth a shot.
gary g.
At 03:40 AM 12/12/2001, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am an undergraduate ChemE major at the University of Rhode Island and I had a question on techniques and AFm capabilities.
Is there a way to measure the force a piezoelectric crystal applies with a certain voltage using an AFM and proscan version 1.5b.0001
Right now I can use the cantiliever to exert force and measure the spring constant of a sample by pushing down, using spectroscopy.
Can it be done without pushing the cantilever down, and instead put a voltage though the crystal and have the crystal push up and measure the force exerted?
Also, on a side note, what is the best way to prepare a blood sample onto a slide (or what would be better than a slide) in a study on the aging of blood. This study will be using primarily AFM techniques, and perhaps nanoindentation. It will also be using the same samples for several weeks, perhaps monthes. After only a week the samples are already crumbling. How can I keep them at ambient conditions, and still keep them from breaking apart?
Hello listers: I am currently trying to prepare TEM samples of polycrystalline strontium titanate (grain size ~20 micron) by conventional dimpling/ion milling. However, due to porosity, I have been having problems in thinning down this material. One of the suggested sample prep methods was to thin the material up to say ~50micron and then embed some sort of epoxy in it, so that the epoxy will fill in the pores. Could someone explain to me how to do this? What type of epoxy should I use and how do I ensure that the epoxy has filled up the pored? Thanks, Prad
Pradyumna Prabhumirashi Dept. of Materials Science Northwestern University Phone: (847)491-7798 Fax : (847)491-7820
A postdoctoral associate position is immediately available in the Department of Molecular & Cellular Biology, Baylor College of Medicine, Houston, TX. The work focuses upon steroid receptor function in an integrating molecular, biochemical, genetic and cell biological approach, and is extremely imaging intensive (including both live and fixed cells, LM/EM). Specific interests address questions of the relationship between transcription factor function, subnuclear organization, intranuclear dynamics and associations with nuclear structure. This single cell approach also allows examining in vivo protein-protein interactions, large-scale chromatin organization, and direct transcription read-out using cell lines with integrated bioluminescent reporters. Imaging resources include state-of-the-art confocal and deconvolution microscopy, and a new digital transmission electron microscope. All microscopes and robust computer/software resources are housed in a completely redesigned/renovated Integrated Microscopy Core. The ideal candidate will have extensive molecular/biochemical experience, OR, extensive imaging/computer training. The ability to communicate exceptionally well (oral and written) and interact with a larger group studying mechanisms of receptor function from many other angles is imperative.
Interested applicants should send an electronic CV and a cover letter expressing their research interests and career plans to:
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References:
Stenoien DL, Nye AC, Mancini MG, Patel K, Dutertre M, O'Malley BW, Smith CL, Belmont AS, Mancini MA. (2001) Ligand-mediated assembly and real-time cellular dynamics of estrogen receptor alpha-coactivator complexes in living cells. Molecular and Cellular Biology. 21:4404-12.
Stenoien DL, Patel K, Mancini MG, Dutertre M, Smith CL, O'Malley BW, Mancini MA. (2001). FRAP reveals that mobility of oestrogen receptor-alpha is ligand- and proteasome-dependent. Nature Cell Biology. 3:15-23.
Stenoien DL, Mancini MG, Patel K, Allegretto EA, Smith CL, Mancini MA. (2000). Subnuclear trafficking of estrogen receptor-alpha and steroid receptor coactivator-1. Molecular Endocrinology. 14:518-34
One quick question.....I am currently performing a general TEM fixation on petunia petals. In our lab, we place plant tissue (once dissected and in fixative) in a vacuum to remove the air. This normally causes the tissue to sink. For these petals, however, I have had them in a vacuum 5 times so far (around 15 minutes each time, or until they "bubble over") and some of them are still not sinking..........
It's almost as if they are hydrophobic. Does anyone know if there is something about petals in general that makes them more difficult to infiltrate with the fixative? I am doing leaves as well and they sank to the bottom just fine.
Does anyone else on this list perform a lot of plant tissue (leaves / petals) fixation? If so, do you place your samples in a vacuum, or use a detergent like Triton X to get the tissue to sediment?
I'm wondering how much damage is caused by placing the tissue in a vacuum as opposed to using a detergent. I know the detergent disrupts membranes, but wouldn't a vacuum have the same effect? Are there other means to get the tissue to sediment? Is one technique better than the other?
I am helping a museum establish a protocol to identify residues on African tribal beads, which is thought to be fecal matter. The sample material will be limited to micrograms and will contain mineral and other contaminants. Can anyone recommend microchemical or micro tests that might be used to differentiate fecal matter?
If the material is dried out, the bugs will probably be dead and uncultivable. Microscopy observation, unless there are specific antibodies against enteric bacteria available for labeling, will be useless. Probably the best test would be a molecular test for E. coli (e.g., PCR).
On Mon, 17 Dec 2001, James Martin wrote:
} Date: Mon, 17 Dec 2001 16:10:05 -0500 } From: James Martin {james.s.martin-at-att.net} } To: microscopy-at-sparc5.microscopy.com } Subject: microchemical tests for fecal matter } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am helping a museum establish a protocol to identify residues on African } tribal beads, which is thought to be fecal matter. The sample material will } be limited to micrograms and will contain mineral and other contaminants. } Can anyone recommend microchemical or micro tests that might be used to } differentiate fecal matter? } } With thanks, } } Jamie } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I have gained the above mentioned microprobe, but it doesn't work yet. It was disassembled in another building, brought over and I have reassembled it to the best of my ability. Kind of a baptism by fire.
The chiller is chilling and the water is coursing around the oil reservoir as well as into the appropriate parts under the column. The oil, used to cool the objective lens, is running from inside that reservoir through new hoses. And the vacuum measures in the 0-5 micro amp range where I'm told it should be (its actually 0.3 at the gun and 0.5 in the column). And the penning gauge measures in the 10^-5 range at the main evacuation manifold - that's good, too.
There is a gauge (Load/Filament) on the Accelerating Voltage panel that should light up "when the vacuum is at the desired level" according to the manual. But something is not allowing that to happen. Consequently, the high voltage cannot be turned on. And there is the rub.
We do not have a service contract on this beast yet. I have called the service people and they have given me a considerable amount of help to date. We just can't get any farther than this. Is there any help for me and my albtross?
Kind regards, --
+++++++++++++++++++++++++++++
R. Ann Bliss Technician, Chemistry and Materials Science Materials Science and Technology Division Lawrence Livermore National Laboratory
Please let me give you a perspective from that of a photographer. In this respect, I have poop specimens which are between days and years old. They all seem to share the same basic characteristics. There is fiber, bulk, stuff, and lots of E. coli. I have also found a few cocci in the mess.
These specimens were sealed under #1 cover slips with cyanoacrylate glue. They are still viable to this day.
The specimens I made for SEM were not very sophisticated. I would have fixed them with Osmium and done CPD. I am not set up to do this. But what I found was that the bacteria were crushed by the high vacuum of the SEM....but they were still viable, morphologically. But without fixing, they deteriorate rapidly. Even stored at 40mT, they do not hold up. Being devoid of Oxygen, the bacteria seem to be confused--as I have seen nearly zero spores. Only dead rods. Of course, there may be other organisms present which do not sporulate. Anyway, not many spores are seen.
It seems to me that the standard protocol for bacterial fixation is quite good and certainly appropriate. In contrast, the LM method can go on for a long time. But of course, the magnification and resolution between LM and SEM is quite different.
But...to your problem. Given that you have a very small amount of material with which to work, experimentation is not a luxury. I would suggest taking a small amount of material and doing a gas chromatograph analysis on it and then taking another small amount and examining it under SEM. With the SEM of the material, you will probably see a lot of crap that is not at all associated with the human crap (feces). What I mean is, junk. This will take some doing to evaluate. Sorting it all out will not be easy at all.
Lots of luck.
gary g.
At 01:10 PM 12/17/2001, you wrote:
} I am helping a museum establish a protocol to identify residues on African } tribal beads, which is thought to be fecal matter. The sample material will } be limited to micrograms and will contain mineral and other contaminants. } Can anyone recommend microchemical or micro tests that might be used to } differentiate fecal matter? } } With thanks, } } Jamie
I would recommend you ask this question to the SPM mailing list http://www.di.com/spm%20forum/spmforummailinglist.html
You may also find the information you need in the application notes found at the DI and Thermomicroscope websites (they are now the same company but still have separate websites): - www.di.com and http://www.tmmicro.com/
DeltaDOT Ltd. PFSG group ACE Building Department of Bioengineering Imperial College London SW7 2AY
0207-594-5174 0794 - 1312335
www.deltadot.com
Information on scanning probe & microfluidics : www.achem.ic.ac.uk/gsanders **************************************************************************** ******* **************************************************************** ----- Original Message ----- } From: "Keith Cochran" {Cochrak-at-egr.uri.edu} To: "Microscopy Listserve" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, December 17, 2001 12:41 AM
Karli Yes, petals are very often hydrophobic. They are covered with a waxy, hydrophobic cuticle. The cell surfaces are very often papillate and the papilla surface may be minutely wrinkled, increasing the water contact angle so that the surface is non-wettable. The most important thing (and this applies to fixation of many types of plant leaves too) is that the pieces you are fixing are small enough for the glutaraldehyde fix to penetrate rapidly via the cut edges - it will penetrate only exceedingly slowly, if at all, via the cuticle, so total immersion is not worth striving for. Cutting pieces 1/2 (half) to 1 mm square should be the aim. Do this using a slicing action with a very sharp degreased razor blade with the petal in a pool of glutaraldehyde in a petri dish, so that cut surfaces are immediately exposed to fix. Subsequent vacuum infiltration will help to get the fix into the tissue if the petals (or leaves) have a lot of intercellular air space, but they will continue to float.
Chris
----- Original Message ----- } From: "Karli Fitzelle" {fitzelle.1-at-osu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, December 17, 2001 8:51 PM
I am DR Nelson Afam {a Civil servant in the ministry of Health} here in Nigeria. I know this letter would come to you as a surprise because we have not met before either physically or through correspondence. I got your contact from our chamber of commerce here in Nigeria and have no doubt in your ability to handle this proposal involving huge sums of money.
The Subject: My father chief Michael Afam {Now late} was the Royal head of my community, Eleme {an oil rich town} in Nigeria. My community produce 5.8% of the total annual crude oil production in Nigeria and 0.5% of the Dollar value of each barrel is paid to my father as royalty by the federal government. In his position as the royal head and chairman of the Akwaeze oil trust fund, he made some money which he behind for his children. The money is twenty five Million, Six thousand US Dollars {$25 .6m}. This money originated from the accumulated royalties between 1998.
Due to poor banking system in Nigeria and political\economics instability as a result of past military rules {1985-1999}, he deposited this money in a “Box” with an open beneficiary in a local security firm. He told the security Company that the content of the box is Gold worth $25.6m. And the box will be safeguarded until he finishes arrangement to transfer it abroad. He was planning this when he died last year of heart Attack.
The Proposal: Just before my father died, he called my attention in confidence to the money and charged me to look for a foreigner who would assist me in the transfer\investment of the fund. So I would be very grateful if you could accept to help me achieve this great object. I promise to give you 20% of total fund s transferred as compensation for your assistants. Five present {5%} would be set aside to take care of all expenses we may incur during the transaction. To the indicate your interest, contact me urgently and confidentially through E-mail for information and the roles you will play in the business. God bless you. DR Nelson Afam
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Jeff- I do not know what beam energy you are using, but another possibility is that the sample(s) in question are charging enough to reduce the effective landing energy of the primary electrons thus altering the peak ratios. I would be surprised if this is what is occurring in your case since the C and F peaks aren't that far apart in energy, but fluorocarbons can be very insulating. With my PGT EDX system in mapping mode, I can view the quasi-real-time count rates in the C and F energy windows. Perhaps if your system can do the same, that could shed some light on your problem (e.g. are both C and F signals getting smaller with the F signal doing so faster, or is F going down as C is going up?). Are the samples gold coated? Does gold coating eliminate the observed effect? Good luck.
Sincerely, Matthew Ervin, Ph.D. (301)394-0017 phone, (301)394-1559 fax MErvin-at-ARL.Army.mil
M/S: AMSRL-SE-RL US Army Research Laboratory 2800 Powder Mill Road Adelphi, MD 20783-1197
Disclaimer: The opinions and views expressed above are those of the author and do not necessarily represent those of the U.S. Army Research Laboratory or any other government agency
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I am currently working on an EDS analysis project on carbon / fluorine matrices. Usually carbon starts as the taller peak, and remains so throughout the acquisition. Occasionally, however, when starting an spectrum acquisition I will see the fluorine peak start as the taller peak, then slow to become the shorter peak.
I was told by an observer that there is a phenomenon known as "chemical shift," that could have an impact on the amount of certain elements in the interaction volume of a sample. This person didn't know much about it other than it could give you inaccurate quant results.
Is there such a thing as chemical shift, and if so, could someone explain what exactly is happening during the process?
Karli, This primary fixative worked very well for Arabidopsis inflourescences even the smallest buds. I also used it for fixing bean (Fava) leaves but in that case I needed a brief vacuum to submerge the leaf samples. The reference is included. 2.8 % glutaraldehyde Buffer 0.1 M Hepes buffer (pH- 7.2) + 0.02% Triton X-100 Buffer wash 0.1 M Hepes buffer
Owen, Heather A., and C.A. Makaroff, Ultrastructure of microsporogenesis and microgametogenesis in Arabidopsis thaliana (L.) Heynh, ecotype Wassilewskija (Brassicaceae) Protoplasma (1995) 185:7-21
It was a long time ago that I worked with 733, but as I remember the vacuum control board has two or three variable resistors which set thresholds, including threshold for turning on high voltage. This board is not very stable and thresholds needs to be readjusted from time to time. I believe it could be your problem. Consult with manual which resistor control high voltage threshold (or are they marked on the board?).
Good luck,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: R. Ann Bliss [mailto:bliss5-at-popcorn.llnl.gov] } Sent: Monday, December 17, 2001 7:06 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: JEOL 733 Help needed } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Happy Holidays: } } I have gained the above mentioned microprobe, but it doesn't work } yet. It was disassembled in another building, brought over and I have } reassembled it to the best of my ability. Kind of a baptism by fire. } } The chiller is chilling and the water is coursing around the oil } reservoir as well as into the appropriate parts under the column. The } oil, used to cool the objective lens, is running from inside that } reservoir through new hoses. And the vacuum measures in the 0-5 micro } amp range where I'm told it should be (its actually 0.3 at the gun } and 0.5 in the column). And the penning gauge measures in the 10^-5 } range at the main evacuation manifold - that's good, too. } } There is a gauge (Load/Filament) on the Accelerating Voltage panel } that should light up "when the vacuum is at the desired level" } according to the manual. But something is not allowing that to } happen. Consequently, the high voltage cannot be turned on. And there } is the rub. } } We do not have a service contract on this beast yet. I have called } the service people and they have given me a considerable amount of } help to date. We just can't get any farther than this. Is there any } help for me and my albtross? } } Kind regards, } -- } } +++++++++++++++++++++++++++++ } } R. Ann Bliss } Technician, Chemistry and Materials Science } Materials Science and Technology Division } Lawrence Livermore National Laboratory } } _____________________________ } }
Regarding the recent question I posted about flashing current decreasing with each tip flash in a Hitachi S4700 FESEM, the problem appears to have been in the gun cable contacts. The Hitachi engineer flashed the tip several times, getting tiny increases in current each time, then checked the contacts on the cable. The next flash produced a MARKED increase in flashing current and the system is now running normally.
My question was aimed more at understanding at the theory behind all this, since the scope was performing just fine, but it turns out to have been a simple mechanical problem fixable in about five minutes.
Thanks again for all the replies.
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
When I open a SE image into Photoshop (v6), it has been my practice to assign a generic gamma=2.2 working space to it. Better, would be to assign whatever space it was acquired with (i.e., the actual display profile for the SEM computer). My thinking is, since we make all adjustments with that display, it is the gamma space I should be associating the SE image to. And indeed, the image appears just like it did at the SEM computer.
However ... the SEM image acuisition software only allows me to fill up the histogram, endpoint to endpoint, without any tools for what's between (gamma). Am I somehow doing the acquisition an injustice with respect to the detector's true gamma?? What if the SE detector's true gamma is unity (e.g., like x-ray maps)?? Has anyone ever determined what a SE detector's apparent gamma is?? If for the sake of the images' data integrity, shouldn't acquisition software be aware of detector gamma, and compensate for the monitor's gamma??
genuinely ... michael shaffer :o) Avalon Peninsula, Newfoundland
I cannot help this person. I was wondering if anyone out their could. Please contact him directly.
-----Original Message----- } From: Nellis, David [mailto:dnellis-at-ncifcrf.gov] Sent: Tuesday, December 18, 2001 11:12 AM To: 'semguy-at-semguy.com' Cc: Giardina, Steve
Dear Listies,
I'm having a bit of trouble sectioning epidermis and dermis. The dermis has thick collagen bundles that tear when thin sectioned for TEM. The epidermis stays intact.
I've tried LR White for better infiltration, initial 15% resin infiltration, extended infiltration times, increased and decreased knife angle, glass knives, diamond knives, decreased and increased cutting speeds.
Befuddled,
Tim Quinn University of Kansas Research Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
I don't have an answer to your specific question, but I do have a paper titled "Technique for the Preparation of TEM Plan-View and Cross-Section Samples of YBa2Cu3O7 Thin Films on MgO and SrTiO3" by Dr. Stephen Strieffer that may be of help. If you would like a copy of the paper, please send me your complete mailing address off-line and I will get it out to you. We also have many other technical papers and application notes available on our website under "Applications Support" at www.southbaytech.com.
DISCLAIMER: South Bay Technology produces equipment and supplies which are described in the above mentioned paper and, therefore, has a vested interest in promoting their use.
I hope this helps.
Best regards-
David
Pradyumna Prabhumirashi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello listers: } I am currently trying to prepare TEM samples of polycrystalline } strontium titanate (grain size ~20 micron) by conventional dimpling/ion } milling. However, due to porosity, I have been having problems in } thinning down this material. One of the suggested sample prep methods } was to thin the material up to say ~50micron and then embed some sort of } epoxy in it, so that the epoxy will fill in the pores. } Could someone explain to me how to do this? What type of epoxy should I } use and how do I ensure that the epoxy has filled up the pored? } Thanks, } Prad } } Pradyumna Prabhumirashi } Dept. of Materials Science } Northwestern University } Phone: (847)491-7798 } Fax : (847)491-7820
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
You have to be careful when referring to gamma. I read a technical article about 1-1/2 years ago that discussed the use of gamma processing and the definition of gamma in different media. It is different for display monitors, film, SEM's etc. I wish that I had photographic memory so that I could recall the points -but I do remember being surprised because of the different definitions in various fields of imaging technology. I had learned about gamma from an SEM course and thought of it as the processing curves shown in the Goldstein et al. book.
I don't think that you should be using gamma processing on SEM images after you collect them. The SEM should have gamma processing on the output of the detector amplifier to the analog to digital converter. Do it electronically to capture what you need from the image. Of course, you should set up the digital image so that as much of the gray scale is used as possible. After a digital image is acquired, the middle control in Photoshop's level control, i.e. the gamma, should only be needed to be moved slightly to adjust the average gray level of the image. If your SEM doesn't have this ability to adjust gamma before the image is digitized, then you can always do a gamma correction using the LUT (Look up table. My philosophy is to use as much of the gray scale as possible when the image is made and to put the average gray level where I like it. I adjust all the display monitors to display what a properly set up image should look like.
Another thing about the SE detector is that the PMT ages. So that if you did set up or relied on some value, over time it would change.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: michael shaffer [mailto:rarewolf-at-roadrunner.nf.net] Sent: Tuesday, December 18, 2001 10:26 AM To: MSA listserver
When I open a SE image into Photoshop (v6), it has been my practice to assign a generic gamma=2.2 working space to it. Better, would be to assign whatever space it was acquired with (i.e., the actual display profile for the SEM computer). My thinking is, since we make all adjustments with that display, it is the gamma space I should be associating the SE image to. And indeed, the image appears just like it did at the SEM computer.
However ... the SEM image acuisition software only allows me to fill up the histogram, endpoint to endpoint, without any tools for what's between (gamma). Am I somehow doing the acquisition an injustice with respect to the detector's true gamma?? What if the SE detector's true gamma is unity (e.g., like x-ray maps)?? Has anyone ever determined what a SE detector's apparent gamma is?? If for the sake of the images' data integrity, shouldn't acquisition software be aware of detector gamma, and compensate for the monitor's gamma??
genuinely ... michael shaffer :o) Avalon Peninsula, Newfoundland
I do not understand purpose of replicating SEM monitor. I think it is more useful to set PC monitor so that it will correspond to printed image (or to averege PC's monitor, if digital images are going to be electronically distributed).
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: michael shaffer [mailto:rarewolf-at-roadrunner.nf.net] } Sent: Tuesday, December 18, 2001 9:26 AM } To: MSA listserver } Subject: SE detector gamma } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } When I open a SE image into Photoshop (v6), it has been my } practice to } assign a generic gamma=2.2 working space to it. Better, } would be to assign } whatever space it was acquired with (i.e., the actual display } profile for } the SEM computer). My thinking is, since we make all } adjustments with that } display, it is the gamma space I should be associating the SE } image to. And } indeed, the image appears just like it did at the SEM computer. } } However ... the SEM image acuisition software only allows } me to fill up } the histogram, endpoint to endpoint, without any tools for } what's between } (gamma). Am I somehow doing the acquisition an injustice } with respect to } the detector's true gamma?? What if the SE detector's true } gamma is unity } (e.g., like x-ray maps)?? Has anyone ever determined what a } SE detector's } apparent gamma is?? If for the sake of the images' data integrity, } shouldn't acquisition software be aware of detector gamma, } and compensate } for the monitor's gamma?? } } genuinely ... michael shaffer :o) } Avalon Peninsula, Newfoundland } } } }
Hello, We want to determine the lattice parameter of nanoparticles on amorphous carbon as a function of size in the TEM. The nanoparticles are smaller than an extinction length, so I can't use dynamical diffraction techniques such as CBED. We are planning on coating the backside of the carbon film with a standard such as gold to use as a reference for SADP analysis. Does anyone know of a better technique to use to determine the lattice parameter of these nanoparticles in the TEM?
Thank you in advance,
Tom Schamp
University of Virginia Dept. of Materials Science cts2v-at-virginia.edu
We are considering purchasing a CL detector for our Philips (now FEIC) XL30 SEM. We are looking for a cost effective CL detector for diagenetic and paragenetic studies of geological materials, typically sandstones. I have looked over the Gatan (formerly Oxford) web site and I wondered if any users out there have used these detectors? If so could you send recommendations or advice regarding the possible options for CL detectors? If there are any other companies out that also make detectors please let me know.
Thanks in advance,
Nick Wilson
Dr. Nick Wilson Research Scientist Geological Survey of Canada 3303-33rd St. N.W. Calgary AB, Canada T2L 2A7 Tel: 403-292-7045 Fax: 403-292-7159 Email: nwilson-at-nrcan.gc.ca
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I think you can try to determine it by means of HRTEM.
Chun-Ming Li Dept. of Materials Science and Engineering University of Illinois at Urbana-Champaign 1304 West Green St. Urbana, IL 61801 Email: lichun-at-uiuc.edu
Tom Schamp wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } We want to determine the lattice parameter of nanoparticles on } amorphous carbon as a function of size in the TEM. The nanoparticles } are smaller than an extinction length, so I can't use dynamical } diffraction techniques such as CBED. We are planning on coating the } backside of the carbon film with a standard such as gold to use as a } reference for SADP analysis. Does anyone know of a better technique to } use to determine the lattice parameter of these nanoparticles in the } TEM? } } Thank you in advance, } } Tom Schamp } } University of Virginia } Dept. of Materials Science } cts2v-at-virginia.edu
We have a group in our institution that is putting together a setup for microinjection. The group interested in this setup is going to be microinjecting fluorescent labeled compounds into adherent cells. We are currently looking at a Zeiss Axiovert 25 CFL. This is a very popular microscope which allow for phase contrast and fluorescence. The only drawback of this microscope and others in this price range is that the mercury lamp is only 50 Watts. The 50W lamp is fine for routine fluorescence but my question is whether it will be effective in picking up dimly lit samples like the fluorescent compounds we are going to microinject?
I received this query, and I cannot help her. Anyone out there who can? Please respond offline to Dr. Marszal.
We wonder if it is possible to have TEM images (with negative staining) taken in your facility as a service. We need to analyze protein polymer samples. If it can be done as a service, what fee could we expect per sample? Such an analysis was done for us before and we know the approximate conditions of the experiment. Also, if it can be done, how soon can we do it? If you do not provide the service, what are the other options; can I participate in the analysis although I have never operated an electron microscope?
Ewa Marszal, PhD FDA/CBER HFM-345 1401 Rockville Pike, Suite 200 N Rockville, MD 20852-1448 phone: 301-402-4638 fax: 301-402-2780
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I'd suggest a harder resin, an epoxy type, to give support to the collagen bundles. Try Spurrs or Embed812, formulated for a harder block. Both are also a fairly low viscosity type resin. You may have to try a few hardness formulas to get it right. Use the diamond knife, if its in good shape, that is.
Good luck,
Gib
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} Dear Listies, } } I'm having a bit of trouble sectioning epidermis and dermis. The dermis has } thick collagen bundles that tear when thin sectioned for TEM. The epidermis } stays intact. } } I've tried LR White for better infiltration, initial 15% resin infiltration, } extended infiltration times, increased and decreased knife angle, glass } knives, diamond knives, decreased and increased cutting speeds. } } Befuddled, } } Tim Quinn } University of Kansas } Research Assistant } Natural History Museum and Biodiversity Research Center } Dyche Hall Room 414 } Lawrence, KS 6604-2454 } 785-864-4556 } tquinn-at-ku.edu
You might want to try HRTEM. Put your samples on amorphous Ge with gold particles. You can use the spots from an FFT of the gold particles to use as a reference standard for your magnification. Then you can use the spots (calibrated from the Au particles) in the FFT from your particles to give you the lattice spacings seen in your particles and count the fringes. Alternatively, you could use the mag from the calculated from the gold particles.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Chun-Ming Li [mailto:lichun-at-uiuc.edu] Sent: Tuesday, December 18, 2001 4:07 PM To: cts2v-at-virginia.edu Cc: Microscopy-at-sparc5.microscopy.com
Hi Tom,
I think you can try to determine it by means of HRTEM.
Chun-Ming Li Dept. of Materials Science and Engineering University of Illinois at Urbana-Champaign 1304 West Green St. Urbana, IL 61801 Email: lichun-at-uiuc.edu
Tom Schamp wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } We want to determine the lattice parameter of nanoparticles on } amorphous carbon as a function of size in the TEM. The nanoparticles } are smaller than an extinction length, so I can't use dynamical } diffraction techniques such as CBED. We are planning on coating the } backside of the carbon film with a standard such as gold to use as a } reference for SADP analysis. Does anyone know of a better technique to } use to determine the lattice parameter of these nanoparticles in the } TEM? } } Thank you in advance, } } Tom Schamp } } University of Virginia } Dept. of Materials Science } cts2v-at-virginia.edu
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} Date: Tue, 18 Dec 2001 20:28:32 -0800 } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: SE detector gamma } } I think that gamma should be set at the computer and its } monitor. Adobe gamma adjust works quite well. There } are hardware units which will do a more precise job but } at higher cost. } } That said, the issue of gamma can take on many meanings } for SE signals. The total dynamic range of a signal is one } thing. The relationship of a peak value to the overall value } range is another. I solve this dilemma by adjusting the } offset of the SE amplifier while independently adjusting the } contrast (SE detector gain). } } A real time histogram (Soft Imaging analySIS ADDA) shows what will } be captured. The contrast and offset can be adjusted to maximize } the tonal range of the input signal. I think this is what other are } suggesting. I do agree. I try to achieve this same goal. If I cannot, } I use Lucis or analySIS' DCE to make up the deficiencies for me. } } gary g. } } } } At 07:26 AM 12/18/2001, you wrote: } } } When I open a SE image into Photoshop (v6), it has been my practice to } } assign a generic gamma=2.2 working space to it. Better, would be to assign } } whatever space it was acquired with (i.e., the actual display profile for } } the SEM computer). My thinking is, since we make all adjustments with that } } display, it is the gamma space I should be associating the SE image to. And } } indeed, the image appears just like it did at the SEM computer. } } } } However ... the SEM image acuisition software only allows me to fill up } } the histogram, endpoint to endpoint, without any tools for what's between } } (gamma). Am I somehow doing the acquisition an injustice with respect to } } the detector's true gamma?? What if the SE detector's true gamma is unity } } (e.g., like x-ray maps)?? Has anyone ever determined what a SE detector's } } apparent gamma is?? If for the sake of the images' data integrity, } } shouldn't acquisition software be aware of detector gamma, and compensate } } for the monitor's gamma?? } } } } genuinely ... michael shaffer :o) } } Avalon Peninsula, Newfoundland
Jamie, It is not clear from your request whether you want to establish that the residue is fecal matter or something else or if you want to identify the individual bacterial species which might be within the fecal matter. It could simply be a matter of placing some of the material into nutrient broth to encourage the growth of whatever was able to survive, realizing of course that if the material is dried out, most organisms would not have survived and therefore would not be able to grow. E. coli is always the predominant bacterial species in fecal matter, but there are also hundreds of other organisms (anaerobic and aerobic).
There are procedures used in the food industry to test for the presence of fecal coliforms (not just E. coli) but they do require viable organisms. They use enrichment procedures to coax out whatever may be present, even if only one organism was able to survive. However, before you can establish any procedures, it would be helpful for you to know why they want this done and what exactly they are looking for.
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----- Original Message ----- } From: James Martin To: microscopy-at-sparc5.microscopy.com Sent: Monday, December 17, 2001 3:10 PM
I am helping a museum establish a protocol to identify residues on African tribal beads, which is thought to be fecal matter. The sample material will be limited to micrograms and will contain mineral and other contaminants. Can anyone recommend microchemical or micro tests that might be used to differentiate fecal matter?
} } I think you can try to determine it by means of HRTEM. } This is very easy to interpret but accuracy is the problem. If you could have small particles of a known structure with a well established lattice parameter on the same grid as your nanoparticles, then it would give you an internal calibration of the magnification. You just can't trust that 500 kX nominal magnification on the TEM is really 500 kX.
Alternatively, if you should be able to determing the parameters using diffraction methods.
Using SAD, the same proviso applies. A standard on the same grid would allow internal calibration. You could use zone axis patterns if you need to determine the structure, or if the structure is already known, you could use a large SA aperture and collect ring patterns. These might be more accurate (especially where the structure is simple such as cubic) since you can calculate the parameters from several different rings and then average them.
With a known structure, it may also be possible using convergent beam methods, using the position of the HOLZ lines in the central transmitted disc. The biggest problem with this is that converging a beam on your nanoparticles may fry them, or at the very least drop lots of carbon contamination from the vacuum on them. So, getting good diffraction patterns is rather difficult. (Cooling the specimen and using zero loss energy filtering can help). Then the lattice parameters are determined by comparison with computer simulated patterns.
The last method is in principle the most accurate, but requires the most work. If you want a quick, but not quite so accurate answer, use either of the first two methods.
I hope this helps
Best wishes
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/ MPI-Stuttgart (in German): http://www.mpi-stuttgart.mpg.de/
I think the point here is two-fold: first, the idea is to collect the best image possible on the SEM, "best image" meaning the image best showing the data being collected, which is not necessarily the most aesthetic image. Second, it's very difficult to determine what the "average gamma of PC monitors" might be -- monitors on Apples generally have a gamma of 1.8, those on Wintel machines something like 2.2, Linux and Unix, I have no clue. Then on top of this, different people adjust the brightness and contrast of their computer's monitors differently. Then there's the differences between CRT and LCD displays. Setting a monitor's display to be the same as a printed display is also interesting. B&W isn't too bad, but color involves color-matching software, meters for measuring colors on the monitor screen, RGB {-} CMYK conversions ... some people get paid lots of money to do this.
Phil
} I do not understand purpose of replicating SEM monitor. } I think it is more useful to set PC monitor so that it will } correspond to printed image (or to averege PC's monitor, if } digital images are going to be electronically distributed). } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } } -----Original Message----- } } From: michael shaffer [mailto:rarewolf-at-roadrunner.nf.net] } } Sent: Tuesday, December 18, 2001 9:26 AM } } To: MSA listserver } } Subject: SE detector gamma } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } } ---------. } } } } } } When I open a SE image into Photoshop (v6), it has been my } } practice to } } assign a generic gamma=2.2 working space to it. Better, } } would be to assign } } whatever space it was acquired with (i.e., the actual display } } profile for } } the SEM computer). My thinking is, since we make all } } adjustments with that } } display, it is the gamma space I should be associating the SE } } image to. And } } indeed, the image appears just like it did at the SEM computer. } } } } However ... the SEM image acuisition software only allows } } me to fill up } } the histogram, endpoint to endpoint, without any tools for } } what's between } } (gamma). Am I somehow doing the acquisition an injustice } } with respect to } } the detector's true gamma?? What if the SE detector's true } } gamma is unity } } (e.g., like x-ray maps)?? Has anyone ever determined what a } } SE detector's } } apparent gamma is?? If for the sake of the images' data integrity, } } shouldn't acquisition software be aware of detector gamma, } } and compensate } } for the monitor's gamma?? } } } } genuinely ... michael shaffer :o) } } Avalon Peninsula, Newfoundland } } } } } } } }
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
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If your fluorescence is to dimly and you are not using plan fluor objectives probably not, you can always use intensified cameras to try to pick most of the fluorescence on the sample
Regards
Alfredo "Seijo, Edward R." {SeijoER-at-moffitt.usf.edu} wrote:
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Have any of you used a Cressington 108 sputtering unit to produce nickel films?? Thus far no one with Cressington here in the US knows if the equipment can do it and I am awaiting England response. Anything I should be aware of?? Thanks
Sorry to be late to the discussion. I assume you want to section the cell itself. Are they naked cells, or covered with coccoliths? Do you care about the cell/lith interface? I ask because I think the real trick is sectioning the soft tissue cleanly while also retaining the mineralized (CaCO3) coccoliths. As I recall, it works with standard fixations and embedding (Spurrs)...and a good diamond knife. Of course, your diamond is at risk.
If your cells are in suspension, it's easiest to pelletize them. As for the whole procedure, I have "seen it done" but no real experience myself! I could get you some references and maybe an experienced contact person.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Saturday, December 15, 2001 10:21 AM, Dee Breger [SMTP:micro-at-ldeo.columbia.edu] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } Hi Jonathan, } } I have no experience with TEM investigation of } coccolithophorids, but for } SEM, the folks here spray them onto stubs - they're so } tiny they adhere on } their own - and then sputter coat. Also, filtering } seawater with coccos in } it and looking at a cutout from the filter paper (in the } SEM, at least) } works well for us. } } Good luck, } Dee } } } } } } Season's Greetings: } } } } I am looking for anyone with experience looking at } } coccolithophorids using } } TEM. } } } } We need to look at sections. The guys are only about 5 um } } in diameter. I } } have done some thin layer embedding and individual cell } } selection before, } } but never on anything this small. I will be checking } } other sources of } } information about techniques, but thought I would ask } } here too, } } } } Any hints or tips on fixing, embedding, etc. that might } } help? } } } } Thanks } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } } ********************************************************** } ***** } Please do not publicly post any of my correspondence } without permission } } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995)
We section human skin in our lab and usually find if the collagen doesn't section it is infiltration. This can usually be traced back to water remaining in the tissue. We use freshly opened 100% etoh in small bottles in the final dehydration steps. Even when using LRWhite which is supposed to be more tolerant of remaining water we have found it best to mix the LRWhite with fresh 100% etoh in the infitration steps leading up to the pure LRWhite.
Or the block of tissue is too big.
Bob Underwood Dermatology Research Center U of Washington
On Tue, 18 Dec 2001, Quinn, Tim Lee wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listies, } } I'm having a bit of trouble sectioning epidermis and dermis. The dermis has } thick collagen bundles that tear when thin sectioned for TEM. The epidermis } stays intact. } } I've tried LR White for better infiltration, initial 15% resin infiltration, } extended infiltration times, increased and decreased knife angle, glass } knives, diamond knives, decreased and increased cutting speeds. } } Befuddled, } } Tim Quinn } University of Kansas } Research Assistant } Natural History Museum and Biodiversity Research Center } Dyche Hall Room 414 } Lawrence, KS 6604-2454 } 785-864-4556 } tquinn-at-ku.edu } }
Still an interesting topic. My current thinking, warped or otherwise, is similar to what I previously posted. But I'd like to clarify a couple of points and expand on some others.
I think that Scott D. Walck's response hit on some good points--in particular, the differences between film, CRTs and detectors. For film, there is a near-straight line relationship between density and log of exposure (log E). A lot of this was discussed much earlier relative to scanners. But anyway, the bottom part of the curve is called the toe, the middle part is called the straight line section, and the top part is called the shoulder (Adams, 1981) The Negative, New York: Little, Brown and Company.
The slope of the straight line portion is a measure of the film's contrast. The slope is the ratio of the density change to the exposure change in the straight line region. This is also called gamma. Thus, gamma equals change in density divided by change in log E in the straight line region. As Adams (1981) points out, a film with higher gamma will have higher contrast than one with lower gamma. This is because a same change in exposure value results in greater density change for the higher gamma film.
The purpose of Adams' Zone system was to be able to adjust the gamma of film to match the range of light for particular scenes. A good example is a scene with blue sky in the upper half of the image and dark forest in the lower half. This could easily represent six to seven stops of light range. The film is typically able to do a decent job within a five stop range. To increase this range, one can combine over exposure, under exposure, over development and under development. The net effect of these actions is to change the slope of the film's curve-- its gamma. The SE detector will have a non-linear characteristic just like film does. So does the scintillator.
In fine art photography, or any other type for that matter, if the film is not exposed (no information on the negative), no other gyrations are going to create information that is simply not there. Thus, I over expose and under develop.
I recently asked this list about shooting microcircuit cross sections which have high Z contrast. The problem here is the same as the bright and dark photographic scene. The solution is a bit different. I actually found two.
First, mix SE and BSE signals as SE-BSE. With high Z, the BSE image is going to be the brightest. Subtracting some amount of BSE from SE will reduce the brightest portion of the image. The other solution is to create a non-linear relationship between input signal and captured signal. I did this using the log G function with my Soft Imaging ADDA active scan system (sorry for the plug). Using the real time histogram, the G value and offset can be set such that the whole 256 or 65536 bit range has information in each bin. And the bright areas will be suppressed in maximum value--thus not saturating the image.
As far as the computer CRT is concerned, I agree completely that it should be calibrated and then left alone. Use Adobe gamma calibrator to do this. Free software and takes about five minutes to set up. There is expensive hardware to do this but it really makes no economic sense for they type of work we are doing with SEM/TEM, etc. I think.
My SEM has a gamma control. It does the same non-linear alteration of input versus capture as previously discussed. Most SEMs probably have gamma controls too. Use them. The basic idea is to set up a log relationship between input and output rather than a linear one. Thus, as input values increase they are reduced in recorded value whereas lower input value are not reduced or reduced very much.
One can still take a rather crappy SEM image and post process it on the computer to make it look much better. There are programs which do this quite nicely. But it is best to get the shot right when it is created, if and when possible.
gary g.
At 07:26 AM 12/18/2001, you wrote:
} When I open a SE image into Photoshop (v6), it has been my practice to } assign a generic gamma=2.2 working space to it. Better, would be to assign } whatever space it was acquired with (i.e., the actual display profile for } the SEM computer). My thinking is, since we make all adjustments with that } display, it is the gamma space I should be associating the SE image to. And } indeed, the image appears just like it did at the SEM computer. } } However ... the SEM image acuisition software only allows me to fill up } the histogram, endpoint to endpoint, without any tools for what's between } (gamma). Am I somehow doing the acquisition an injustice with respect to } the detector's true gamma?? What if the SE detector's true gamma is unity } (e.g., like x-ray maps)?? Has anyone ever determined what a SE detector's } apparent gamma is?? If for the sake of the images' data integrity, } shouldn't acquisition software be aware of detector gamma, and compensate } for the monitor's gamma?? } } genuinely ... michael shaffer :o) } Avalon Peninsula, Newfoundland
No commercial interest, just a satisfied customer.
Sara Miller
On Wed, 19 Dec 2001, Greg Martin wrote:
} Date: Wed, 19 Dec 2001 09:15:09 -0500 } From: Greg Martin {gvm-at-aretha.jax.org} } To: Microscopy-at-sparc5.microscopy.com } Subject: Third-party service for Leica LMs } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hey Folks -- } } Can anyone recommend a good outfit for servicing Leica light } microscopes in the northeast? Please reply to me directly at gvm-at-jax.org. } } Thanks! Greg. } } Greg Martin } Light/Confocal Microscopy } Biological Imaging Service } The Jackson Laboratory } 600 Main Street } Bar Harbor ME 04609 } 207-288-6310 } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
To everyone who responded, thank you. Dr Marszal has contacted me to let me know that she found a lab near her, in Maryland, to do the work. Happy Holidays and a peacefull New Year to you all.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi Ed, I just got off Google where I was surfing through lunch having searched for "microinjection confocal". Thus, I came back to email already loaded with a response for your question, and , of course, much more than you wanted. You might try to contact someone at USF-physiology to get some near-home advice. E.g., Dr. Yip: dyip-at-hsc.usf.edu. I found him by searching USF for "confocal". Please don't think me a pompous git (as Ron Weasely would say) for responding this way, but I can't imagine why you would want to expend the energy to set up a system that didn't fully image the object. I am in possession of an Olympus FV-300 which was acquired for between 145 and 155k$. Remember that you will be operating on specimens in an aqueous environment and that you should look to water immersion objectives whether you use an inverted or upright microscope. They exist having been designed for non-coverglass immersion (upright) or coverglass immersion (inverted). The high res water immersion from Olympus was quoted at around 9-10k$. WOW!!! In any case, if you can find a confocal on which there is time available, then you might be able to pay your way by supplying the water immersion objective - something that many purchasers don't miss until after they have made the deal. Then again, you might get really lucky. Finally, if you really don't need a confocal, then you must get advice from the apps folks at Molecular Probes about the excitation of the fluorescent probes you want to use and their 'exciting' characteristics and how they 'quench' when lit, and how to limit their 'quenching' - when lit. After you get that done, you should demand a chance to test the scope, objective(s) and illumination. Oh yes, I'm not privy to the Zeiss you mentioned, since our inverted, and quite stable, inverted scope is from OLYMPUS, but whatever the scope, insure that the stage doesn't move or that the micromanipulators are stage-mounted. Twenty years ago, I would not have tried an Olympus dealer, but I would now (and my only relationship with them is as a user). Advisory! One spends money for a microscope, because one needs optical and mechanical stability. Without one or the other, even the little funds end up wasted. Nikon made a really solid inverted scope twenty years ago - just perfect for physiologic experiments. I would bet that one could be found on the used market. Its only deficiency, in today's environment, would be that it wasn't infinity corrected. Tube-length corrected lenses are still made, however, and can be acquired for less than 9-10k$. Such optics are usually quite good.
Two really good little books: "Confocal Laser Scanning Microscopy", Sheppard and Shotton, Springer-Verlag, ISBN: 0 387 91514 1. ~$35.00 "Flow Cytometry - first Principles", Givan, A., Wiley-Liss, ISBN: 0-471-56095-2
Also, some bigger tomes,
Methods in Cell Biology, vol. 33 for fluorescent methods, and Methods in Enzymology, vol. 307(under $90), and "Electronic Light Microscopy", Shotton, D., Wiley-Liss, ISBN: 0-471-56077-4.
Hope some of this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Seijo, Edward R. } Sent: Tuesday, December 18, 2001 4:04 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Need feedback on inverted microscopes for microinjection } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have a group in our institution that is putting together a setup } for microinjection. The group interested in this setup is going to be } microinjecting fluorescent labeled compounds into adherent cells. We are } currently looking at a Zeiss Axiovert 25 CFL. This is a very popular } microscope which allow for phase contrast and fluorescence. The only } drawback of this microscope and others in this price range is that the } mercury lamp is only 50 Watts. The 50W lamp is fine for routine } fluorescence } but my question is whether it will be effective in picking up dimly lit } samples like the fluorescent compounds we are going to microinject? } } Any thoughts? } } Thanks. } } } } } } } }
After a quick poll of my technical people here it seems that typical planar magnetron sputtering units, even with a high vacuum system, will not sputter Nickel. This is due to the magnetic properties of the Nickel target material which interferes with the planar magnetron permanent magnet and will not effectively make a suitable thin film. But, you can prepare precise nickel thin films in an ion beam sputtering system such as our IBS/e. If you would like more information on Ion Beam Sputtering, I'd be pleased to send it to you or you can visit our website at www.southbaytech.com and type in the keyword "IBS/e".
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
Original Message ----- } From: Scott Whittaker {Whittaker.scott-at-nmnh.si.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, December 19, 2001 11:41 AM
Hello people.......
I´m looking for a Objetive Stage High Resolution for a TEM Philips EM 301..... any other spare parts are welcome.....
If any knows Santa Claus, please contact me via email.......
merry christmas.......
Gabriel Adriano Rosa Centro de Imagenes y Microscopia Departamento de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail cimic-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
During my last job, I had no problem sputtering Ni or NiFe alloys with a magnetron source. I used a Sputtered Films R&D System, that employs a conical magnetron source. The magnetic field from the gun (source) magnets will interact with the magnetic target material, but with a sufficient source to substrate distance, high-quality, uniform Ni films can be achieved. IBS will undoubtedly work, but the deposition rate is inherently slow and the technique is much more expensive than magnetron sputtering. IBS is generally used where thickness control is extremely critical, like giant magnetoresistive multilayer applications.
Andrew Tzanavaras, M.S. Engineering Manager Peripheral Storage, Inc. USA
David Henriks wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Scott: } } After a quick poll of my technical people here it seems that typical } planar magnetron sputtering units, even with a high vacuum system, will } not sputter Nickel. This is due to the magnetic properties of the Nickel } target material which interferes with the planar magnetron permanent } magnet and will not effectively make a suitable thin film. But, you can } prepare precise nickel thin films in an ion beam sputtering system such } as our IBS/e. If you would like more information on Ion Beam } Sputtering, I'd be pleased to send it to you or you can visit our } website at www.southbaytech.com and type in the } keyword "IBS/e". } } DISCLAIMER: South Bay Technology produces equipment and supplies as } described above and, therefore, has a vested interest in promoting their } use. } } Best regards- } } David } } -- } David Henriks TEL: +1-949-492-2600 } South Bay Technology, Inc. FAX: +1-949-492-1499 } 1120 Via Callejon } San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com } } *** www.southbaytech.com *** } } Original Message ----- } } From: Scott Whittaker {Whittaker.scott-at-nmnh.si.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, December 19, 2001 11:41 AM } Subject: Nickel thin film production } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } Have any of you used a Cressington 108 sputtering unit to produce } nickel } films?? Thus far no one with Cressington here in the US knows if the } equipment can do it and I am awaiting England response. Anything I } should be } aware of?? Thanks } } } }
Also, beware that nanoparticles below a certain size (in the realm of a few nanometers) are prone to certain image abberations, which leads to somewhat erroneous lattice measurements (I believe up to 10-20%). I wish I had the reference and more concrete info, I apologize. It was a talk at the Spring American Chemical Society Meeting by a microscopist at Monsanto. I unfortunately threw away the program! Anyone else familiar with such findings?
Paul E. Anderson, Ph.D. Department of Chemistry Northeastern University Boston, MA
In a message dated 12/19/01 3:14:22 AM Eastern Standard Time, lichun-at-uiuc.edu writes:
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The term of "chemical shift" is defined as the energy peak shift on certain condition such as oxidization status. Compare to the pure element, its oxidized molecule would have an energy peak shift due to chemical bonding (with O, F etc). It is on the order of 0.5-3 eV and won't be "seen" by any current EDS detector. This is not the case for the F intensity low, because the chemical shift has nothing to do with the intensity.
What you have seen on the F peak lower in intensity depends on the material property. Some F-polymers are very sensitive for E-beam called beam damage. The electron charge from e-beam can destroy the bounding on certain points of molecule chain and causes it broken. The F fragment such as CF, CF2 and CF3 (F comes with C) can be released as gas and goes away from the vacuum. After certain period of time, the F compound will be gone and no more F can be determined on the surface of test point. Also there will be C-rich material left on the site causing C peak growth. No all F-polymers are sensitive for e-beam (Teflon is the stable one). I use EDS for F-polymer lubricant analysis (used on the surface of hard drive disks) and I can see this phenomenon clearly. There is impossible to do quantitative analysis for this kind of material.
Zhiyu Wang, Sr. Engineer Maxtor Corp. Milpitas CA
----- Original Message ----- } From: "Matt Ervin" {mervin-at-arl.army.mil} To: "Oakley, Jeff" {oakleyj-at-rayovac.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, December 18, 2001 2:08 PM
ACEM 17 The 17th Australian Conference on Electron Microscopy Adelaide Convention Centre, Adelaide, SA February 4 - 8, 2002
It's not too late to book a winter escape trip - why not combine an exciting conference and summer holiday in Adelaide, South Australia in Feb 2002! Registrations will be accepted right up until the conference date, and there are a range of accommodation choices suitable for all.
Encompassing the areas of Electron, Scanned Probe and Confocal Microscopies, this important meeting will provide an opportunity to explore the many applications of these techniques, to keep abreast of the latest developments and to meet and mix with many of Australia's and the world's leaders in the field.
Invited and Keynote Speakers include: Martin Bootman; Confocal (biological), David Cockayne; Nanostructure, Patrick Echlin; Analytical Microscopy of Biomaterials, Mark Fricker; Confocal (biological,) David Jefferson; Emerging Techniques, Jan Leunissen; Immunolabelling, Sergei Magonov; SPM, Ohad Medalia; Cellular Tomography, Greg Meeker; Microbeam Analysis, Sara Miller; Virology, Daniel Mueller; SPM - (biological), David Williams; Microanalysis, Nestor Zaluzec; Analytical Microscopy/Telepresence, Jamie Chapman; Reproductive Biology, Jacques Dubochet; Cryotechniques, Alex Hyatt; Virology/Cryotechniques, Mary, Mah-Lee Ng; Virology/Confocal
Workshops A series of pre-conference workshops will be held to allow participants to gain hands-on experience in the latest techniques and applications. Many of the invited and keynote speakers will participate as presenters, providing a wonderful opportunity for interaction with these noted scientists. Some of the workshops are close to full - book soon!
Cryo Scanning Electron Microscopy Patrick Echlin, John Terlet An introduction to SEM low-temperature techniques such as specimen preparation (freezing methods), field-emission SEM cold stage operation, obtaining optimum imaging conditions and image interpretation.
Environmental Scanning Electron Microscopy, Techniques and Applications Brendan Griffin, FEI representative, Matthew Phillips The Workshop will cover all aspects of the theory and practice of environmental scanning electron microscopy (ESEM). The use of ESEM for a number of applications will be shown with hands-on demonstrations.
Image Analysis Allan Jones, Arne Muller, T.K. See Tho, Craig Noble The first day of this 2 day Workshop will comprise lectures and practicals that cover the major aspects of image analysis. The lectures will cover the range from basics such as image capture and digitisation to the latest techniques and algorithms for image processing. The practicals will be hands-on problem solving sessions. Day 2 will be an open forum in which participants will be able to discuss analysis strategies for images relating to their particular interests and to discuss current technologies and future directions with vendors of image analysis software packages.
Leica-Aurion Workshop on Cryotechniques and Immunolabelling Ross Boadle, Jacques Dubochet, Alex Hyatt, Jan Leunissen, Ian Lamswood, Alasdair McDowall, OhadMedalia & Graham Tranter The Leica-Aurion Workshop will be a major event of ACEM17. This Workshop brings together a number of experts in the field of immunolabelling and cryotechniques. The Workshop is based on Leica Microsystems Workshops generally available only in Europe as well as other courses presented locally and overseas. This comprehensive Workshop will cover all aspects of immunolabelling and cryo-preparation techniques, including staining, thin sectioning, immuno-histology, gold labelling, immunopathological techniques, cryo-sectioning and cryo-staining.
Light Element Analysis: Problems and Solutions Greg Meeker, Colin MacRae The Workshop will focus on light element analysis issues such as sample preparation, soft x-ray spectroscopic strategies and techniques, calculation of density of states and matrix corrections.
Quantitative Confocal Microscopy Martin Bootman, Guy Cox, Mark Fricker, Leann Tilley, Meredith Wallwork The Workshop will focus on quantitative confocal microscopy techniques. Formal presentations will be followed by sessions on the confocal and multi-photon microscopes using techniques such as live-cell imaging, 3-D and 4-D techniques, multi-channel imaging and fluorescence recovery after photobleaching (FRAP).
Scanning Probe Microscopy Sergei Magonov, Daniel Mueller, John Thomas The full day workshop consists of 3 parts: 1. An Introduction to Scanning Probe Microscopy will cover the history/development, principles and basic techniques (contact mode, tapping mode, phase imaging, force distance measurements). 2. Application of SPM to biological systems including sample preparation, imaging techniques, information obtainable etc. 3. SPM studies of polymers will highlight the techniques used and information available with a focus on studies at elevated temperatures. A scanning probe microscope system will be used for demonstration. Registrants are asked to indicate their interest in the 3 topics above. The convenor may rearrange the schedule and the time allotted accordingly.
Trace and Major Quantitative X-ray Mapping (a Hands on Approach) Peter Lloyd, Ken Moran The Workshop will present a practical course on aspects of x-ray mapping. Topics will include: instrumentation; how to get your equipment stable: how to get as many counts as possible; x-ray maps using ED and WD; full spectrum mapping; trace element determinations; statistical benefits.
Virology Using Electron & Light Microscopy Sara Miller, David Howell, Alex Hyatt, Marilyn Henderson The virology workshop will cover techniques for preparing samples for examination, including histological, immunohistochemical, negative staining, and thin sectioning methods for virus detection. It will describe the pathology, cytology and ultrastructural characteristics of infections. It will also demonstrate a "key" for morphologically identifying viruses down to the family or genus level so that an atlas can be easily consulted.
The Trade Exhibition will be a comprehensive and exciting display of the latest equipment and accessories for microscopy, imaging and analysis. We are most grateful to our sponsors and exhibitors for their very generous support of ACEM17.
Sponsors
Platinum: FEI Company, Leica Microsystems, OED P/L (LEO), Soft Imaging System GmbH, Thomson Scientific Instruments (Gatan)
Gold: JEOL (Australasia) P/L
Social Functions: FEI Company (Conference Dinner), Thomson Scientific Instruments (Gatan) (Welcome Reception), Oxford Instruments (Zoo-B-Que)
Session Sponsors: Adelaide University, South Australian Centre for Manufacturing
Poster Competition: ProSciTech
Micrograph Competition: Unibooks
Workshop Sponsors: Adelaide University, Aurion, Bio-Rad Laboratories P/L, Carl Zeiss P/L, Digital Instruments Inc, FEI Company, Group Scientific P/L, Leica Microsystems P/L, Moran Scientific P/L, Olympus, Soft Imaging System GmbH
As is traditional at all ACEMs, Poster and Micrograph Competitions will be run. Categories for the Poster Competition will be Best Biological Poster, Best Physical Poster and Best Student Poster. Categories for the Micrograph Competition are SEM, TEM, SPM and Confocal. Entries will be judged on aesthetic, photographic and scientific merit. Entry details are available on the website.
Visit the website often for regular updates! http://www.adelaide.edu.au/CEMMSA/acem17
We have an Ion Tech (later called Atom Tech, last known as MECA2000 in France) magnetron sputterer that has been modified to sputter Fe, Ni etc. We also bought an AJA magnetron head for magnetic materials.
The Ion Tech magnetron has a ring of small magnet stacks around the circumference and a central magnet stack of the opposite polarity to produce the magnetic field. We weakened the central stack (replaced rare earth magnets with an Fe rod) to allow magnetic materials to be sputtered. We also use thin magnetic targets (0.5 to 1mm thick compared to 3mm of non magnetic materials).
Not being a sputtering expert I'm rusty on the exact details but I could check it out if you want to follow it up.
Regards, Ron
On Wed, 19 Dec 2001 18:29:04 +0200 Andrew Tzanavaras {attz-at-postoffice.pacbell.net} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Scott, } } During my last job, I had no problem sputtering Ni or NiFe alloys with a } magnetron source. I used a Sputtered Films R&D System, that employs a } conical magnetron source. The magnetic field from the gun (source) magnets } will interact with the magnetic target material, but with a sufficient } source to substrate distance, high-quality, uniform Ni films can be } achieved. IBS will undoubtedly work, but the deposition rate is inherently } slow and the technique is much more expensive than magnetron sputtering. } IBS is generally used where thickness control is extremely critical, like } giant magnetoresistive multilayer applications. } } Andrew Tzanavaras, M.S. } Engineering Manager } Peripheral Storage, Inc. } USA } } David Henriks wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Scott: } } } } After a quick poll of my technical people here it seems that typical } } planar magnetron sputtering units, even with a high vacuum system, will } } not sputter Nickel. This is due to the magnetic properties of the Nickel } } target material which interferes with the planar magnetron permanent } } magnet and will not effectively make a suitable thin film. But, you can } } prepare precise nickel thin films in an ion beam sputtering system such } } as our IBS/e. If you would like more information on Ion Beam } } Sputtering, I'd be pleased to send it to you or you can visit our } } website at www.southbaytech.com and type in the } } keyword "IBS/e". } } } } DISCLAIMER: South Bay Technology produces equipment and supplies as } } described above and, therefore, has a vested interest in promoting their } } use. } } } } Best regards- } } } } David } } } } -- } } David Henriks TEL: +1-949-492-2600 } } South Bay Technology, Inc. FAX: +1-949-492-1499 } } 1120 Via Callejon } } San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com } } } } *** www.southbaytech.com *** } } } } Original Message ----- } } } From: Scott Whittaker {Whittaker.scott-at-nmnh.si.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Wednesday, December 19, 2001 11:41 AM } } Subject: Nickel thin film production } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } Have any of you used a Cressington 108 sputtering unit to produce } } nickel } } films?? Thus far no one with Cressington here in the US knows if the } } equipment can do it and I am awaiting England response. Anything I } } should be } } aware of?? Thanks } } } } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I should like to inform you, that the list of microscopy meetings for the year 2002 at so called Petr's Microscopy Resources has been extended. You can see it at the
Petr Schauer +---------------------------------------------------------------------+ | Dr. Petr Schauer tel.: (+420 5) 41514313 | | Head of Electron Microscopy Laboratory fax : (+420 5) 41514404 | | INSTITUTE OF SCIENTIFIC INSTRUMENTS (+420 5) 41514402 | | ACADEMY OF SCIENCES OF THE CZECH REPUBLIC e-mail: petr-at-isibrno.cz | | Kralovopolska 147, CZ-612 64 Brno petr-at-dp.fce.vutbr.cz | | Czech Republic www: http://www.petr.isibrno.cz/ | +---------------------------------------------------------------------+
Is anybody else receiving daily junk emails from this list. These range from offers of money to offers of sex and more!!!
The "to" box of the emails is the group email address and I've only been recieving them since I joined this list. Is there any way to stop them other than setting a mail rule for all messages from this group which would be silly as i would miss out on all the emails!
Thanks
Steve Thomas
********************************* Dr Steve Thomas School Of Biosciences The University of Birmingham Edgbaston Birmingham B15 2TT 0121 414 5573 Email: S.Thomas-at-bham.ac.uk
There is also an error component in lattice parameter measurements due to tilt of the particles away from the exact Bragg condition. Think of the situation in reciprocal space, where the reflections are in general elongated. Depending on particle tilt, the Ewald sphere cuts the streaked reflection somewhere along its length and the answer you get - the measured d-spacing - depends on where the relrods are cut.
There was a nice talk by Pete Crozier et al at the M&M conference '01 (page 1078 of proceedings) which deals with this topic. In it is a reference to an article in J. Electron Microscopy 48 (1999) p. 1015. I would imagine you could get more details on this there.
Regards, Wharton
} -----Original Message----- } From: "Pea9000-at-aol.com"-at-sparc5.microscopy.com } [SMTP:"Pea9000-at-aol.com"-at-sparc5.microscopy.com] } Sent: Wednesday, December 19, 2001 8:49 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: What is the best technique to determine nanoparticle } lattice parameters ... } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Also, beware that nanoparticles below a certain size (in the realm of a } few } nanometers) are prone to certain image abberations, which leads to } somewhat } erroneous lattice measurements (I believe up to 10-20%). I wish I had the } reference and more concrete info, I apologize. It was a talk at the Spring } } American Chemical Society Meeting by a microscopist at Monsanto. I } unfortunately threw away the program! } Anyone else familiar with such findings? } } Paul E. Anderson, Ph.D. } Department of Chemistry } Northeastern University } Boston, MA } } In a message dated 12/19/01 3:14:22 AM Eastern Standard Time, } lichun-at-uiuc.edu } writes: } } { { cts2v-at-virginia.edu } }
The answer is no, in general, but a few slip through. For example 2 slipped throught the fliters last night, and doing a trace I see that they originated via 2 automated reply sites which then used some public software list which subsequently sent the messages to this listserver. This Server attempts to filter most of this stuff out but only when the culprits, phrases, keywords etc.... get added to the filter system databses. I monitor things and when the occasional one gets through I add the keywords/phrases to the "database" , so the same route should, in principle, never occur again. No filter is perfect and some will get through.
Just to give you an order of magnitude ~ 100 junk mail messages are rejected / week. So the one or two that slip through represent only the tip of the iceberg.
A general consensus of the info received suggests ideal fixation and embedment conditions for skin would be-
1.Use only unopened ETOH for final 100% dehydration steps. And possibly increase amount of 100% ETOH steps.
2. Use Spurrs resin, maybe adjusted to be slightly harder than normal. Increase infiltration times substancially.
3. Make block face smaller. Cut through epidermis first (least resistance).
I'll be working with blue monkey scrotum and blue mandrill face skin samples. I'll let everyone know how the results turn out.
Thanks to all. Tim Quinn University of Kansas Research Assistant Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
I wanted to thank all who replied to my query on oil backstreaming into the chamber of SEM's, and to provide a summary of the responses.
Owners of multiple makes/models of SEM's replied, and the impression was that this is a very common problem.
The majority of repliers, but not all, indicated that the oil was probably coming from the backing pump. It was indicated that this can be easily tested by IR, which will detect if it is silicone or hydrocarbon. Unfortunately, I cleaned the detector already so that will have to wait for possible future occurrences.
About cleaning the detector, it was pointed out that this can be done by removing the collimator, cleaning that with acetone/methanol, and dribbling methanol over the head of the detector. However, beware! It was by doing this that I somehow caused the thin window to fail. In short, this process should be avioded if possible as it can shorten the life of a thin polymer window, or the adhesive which seals it. If you are doing this frequently, you should act now to get rid of the backstreaming.
Some responders indicated that DP oil can separate into two components at advanced age (which is the case for this instrument). The lighter component can then backstream into the chamber, i.e. it is possible that this is occurring. This is supported by one response which claimed that the mechanical pump had been replaced with a scroll pump and the DP left alone - the oil problem didn't go away.
Several proposed solutions
1) Install a foreline trap and keep it maintained 2) Cut two holes in a refrigerator and insert foreline through it so that trap is inside frig (!) 3) Eliminate oil-based system entirely and go to an oil-free turbo/scroll system. 4) Install a system such as SEM-clean (XEI Scientific), essentially injecting a slow stream of dry nitrogen into the foreline to produce a constant gas flow, which will block oil from backstreaming. 5) Change the switchover from rouging to diffusion pumping to a lower pressure. If the pressure in the chamber is above 100 millitor when the switchover occurs this will be too much for the DP and will cause turbulent flow in the DP leading to backstreaming. 70 millitor was recommended. It was noted that this is a problem on the Hitachi S570. 6) Check cooling lines to cold trap to ensure it is working (we don't have one). 7) Have the detector pumped out so that it will insulate better and the tip won't be cold (eliminates condensation). 8) Get a better mechanical pump. (Our mechanical pump was in fact getting rather hot - too hot to touch - when running continuously, which I gather is a characteristic of that particular pump). 9) Get Will Bigelow's book and learn more about this problem.
So what do you do when you don't have unlimited time to fiddle or money to spend? Do what is easiest and cheapest and hope it works.
This eliminates #2, 3, and 4. I have installed the foreline trap (bakeable). I also found a better mechanical pump, had it serviced and installed it, also changing all the rubber hose. By default I am doing #7 because the detector needs service. I know I should do #9. Number 5 is very interesting and I intend to mention it to our service people to see if it can easily be accomplished.
My advice - If you are buying a new SEM, an oil free vacuum system is a very good idea. Right now I am going to wait and see what I get from the improvements already made. I will let you know how it goes.
Sorry if this is repeat information, I deleted the other "EDS chemical shift" emails.
The microcalorimeter being developed with the support of NIST represents hardware capable of yielding chemical information from x-ray micro-analysis. If and when this becomes a commerical product it will have the energy resolution needed to provide "chemical shift" type information.
For those interested in the microcalorimeter project here is a websit link} http://www.boulder.nist.gov/div814/div814/news/microcal/microcal.htm
Regarding beam damage to fluorinated polymers, it is a fact of life. Every system has a critical dose that is a function of the area of the beam and the current. You can reduce the damage rate by rastering over a larger area, if that is an option. You can check for this effect by acquiring spectra over time and plotting a ratio of the components (say F:C ratio). You can then even assign a "damage parameter" based upon the data exponents. A higher voltage for a given current will tend to reduce the damage rate also, because the same current is distributed over a larger area.
Regards, Ed Principe Defect & Thin Film Characterization Lab Applied Materials Santa Clara, Ca.
"Zhiyu Wang" {zhiyuw-at-attbi.com} on 12/19/2001 01:31:19 PM
Please respond to "Zhiyu Wang" {zhiyuw-at-attbi.com}
The term of "chemical shift" is defined as the energy peak shift on certain condition such as oxidization status. Compare to the pure element, its oxidized molecule would have an energy peak shift due to chemical bonding (with O, F etc). It is on the order of 0.5-3 eV and won't be "seen" by any current EDS detector. This is not the case for the F intensity low, because the chemical shift has nothing to do with the intensity.
What you have seen on the F peak lower in intensity depends on the material property. Some F-polymers are very sensitive for E-beam called beam damage. The electron charge from e-beam can destroy the bounding on certain points of molecule chain and causes it broken. The F fragment such as CF, CF2 and CF3 (F comes with C) can be released as gas and goes away from the vacuum. After certain period of time, the F compound will be gone and no more F can be determined on the surface of test point. Also there will be C-rich material left on the site causing C peak growth. No all F-polymers are sensitive for e-beam (Teflon is the stable one). I use EDS for F-polymer lubricant analysis (used on the surface of hard drive disks) and I can see this phenomenon clearly. There is impossible to do quantitative analysis for this kind of material.
Zhiyu Wang, Sr. Engineer Maxtor Corp. Milpitas CA
----- Original Message ----- } From: "Matt Ervin" {mervin-at-arl.army.mil} To: "Oakley, Jeff" {oakleyj-at-rayovac.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, December 18, 2001 2:08 PM
Examples of such errors are discussed in the following: "Deceptive "Lattice Spacings" in High-Resolution Micrographs of Metal Nanoparticles", J.-O. Malm and M.A. O'Keefe, Ultramicroscopy 68 (1997)13-23. "Fresnel effect in high resolution TEM imaging of small particles", Velimir Radmilovic and Michael A. O’Keefe, in 53rd Ann Proc. MSA, Kansas City, Missouri (1995) 564-565. "Application of cross-correlation technique for the analysis of Fresnel effect in small particle HREM imaging", V. Radmilovic, R. Kilaas, and M.A. O'Keefe, in Proc. European. Congress for Electron Microscopy II (1996) 50-51. "The effect of crystal tilt on high-resolution micrographs of small metal particles", Jan-Olle Malm and Michael A. O'Keefe, SCANDEM-93 (Extended Abstracts of the 45th Annual Meeting of the Scandinavian Electron Microscopy Society), Lund, Sweden (1993) 131-132. Please contact me with further questions. -Michael A. O'Keefe NCEM, LBNL
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Also, beware that nanoparticles below a certain size (in the realm of a few } nanometers) are prone to certain image abberations, which leads to somewhat } erroneous lattice measurements (I believe up to 10-20%). I wish I had the } reference and more concrete info, I apologize. It was a talk at the Spring } American Chemical Society Meeting by a microscopist at Monsanto. I } unfortunately threw away the program! } Anyone else familiar with such findings? } } Paul E. Anderson, Ph.D. } Department of Chemistry } Northeastern University } Boston, MA } } In a message dated 12/19/01 3:14:22 AM Eastern Standard Time, lichun-at-uiuc.edu } writes: } } { { cts2v-at-virginia.edu } }
Actually Nikon has Infinity optics and longer working distances than any other maker, TE300 or TE2000 (visit Nikon USA website) are as solid as the old TMD and these new microscopes has water immersion objectives and an upright microscope with the stand fixed. The actual focusing is done through the nosepiece the N. A. for 60X Water immersion objective is 1.4 as far as I can remember
Alfredo
"Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
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Hi All: I was asked to look at an old SEM we have and I am looking to see if there is company support for an old ISI SEM. I guess thay have become Topcon ? Do they support older ISI SEM's. Let me know if you have any knowledge. Thanks in advance. Thx, Mike Coviello UTA
Dear collectors of valuable stuff, One of my customers is looking for a second stage and door ass'y for his AMRAY 1610. His present one has gotten "hot" and he would like to have a clean one for most of his work. Makes life simpler.
Contact me off line and I can put you in touch with him if you have something.
Thanks and Happy Holidays, Ken Converse owner Quality Images third party SEM service Delta, PA
We had noticed some time ago that our carbon adhesive tabs seemed to develop a series of fine cracks (looks like drying mud) after being sputter-coated. This was not a real problem, just a minor aesthetic annoyance when they sometimes showed up in the background of a micrograph. However, when a client recently expressed concern about these cracks and wondered if sputtering was also damaging her samples, I did a rather unscientific series of tests to see what was going on.
If anyone is interested, we discovered that sputtering at 20mA for 60 secs invariably caused cracking of the tabs, but sputtering at 5mA for 60 seconds did not. Sputtering for 20 seconds at 20mA also avoided the cracks. To see if heating was the culprit, we put an uncoated tab on an SEM stub into a 60 degree oven for an hour. No cracks. We also ran one through the sputter coater cycle, but did not actually apply a coating, to see if the turbo-level vacuum alone would do it. No cracks.
So, if cracked tabs are keeping you awake at night, use a lower sputtering current or a shorter time. If you couldn't care less, well, have a Merry Christmas anyway and delete this!
Many thanks for the useful information received from Drs Hunt, Boucher, Wieland and Quinn. We were able to service the vac valve of the right mill. We replaced the both 'o' rings. The bellows turned out to be good, i.e., no leak there. Also thanks to Dr Mitro for the detailed instructions. Vacuum levels are really good now.
Merry Christmas to all,
---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jearanai-at-nsrc.or.th) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, December 21, 2001 at 02:06:36 ---------------------------------------------------------------------------
Mike - Topcon discontinued service support years ago. We have an ISI (Topcon) SEM that we continue to have serviced by our former ISI service engineer, who is independent now. His name is Chuck Humphrey and his company's name is Scanning Solutions. He has always done an excellent job for us. You can reach him the following ways: telephone 407 234 0676 fax 407 277 4423 email cchumph-at-attglobal.net
Good luck with the scope.
Sincerely,
Jill
Jill Verlander Reed, D.V.M. Associate Scientist Director,Electron Microscopy Core Facility UF College of Medicine P.O. Box 100215 HSC 1600 SW Archer Rd. Room RB-167 Gainesville, FL 32610-0215
-----Original Message----- } From: Mike Coviello [mailto:coviello-at-mae.uta.edu] Sent: Thursday, December 20, 2001 4:58 PM To: verlaj-at-mail-cs.med.ufl.edu
Hi All: I was asked to look at an old SEM we have and I am looking to see if there is company support for an old ISI SEM. I guess thay have become Topcon ? Do they support older ISI SEM's. Let me know if you have any knowledge. Thanks in advance. Thx, Mike Coviello UTA
I guess I should clear some things up here. I am actually impressed with the rapidity and thoroughness of the response from both you and the England group, I was just simultaneously polling the Listers for their experiences, which I feel is the most rapid and effective way to get the nitty gritty tips and info you will never find in any manual or instruction sheet and save several hours of trial and error work. Happy holidays to all.
Scott Whittaker SEM Lab Manager/Senior Dishwasher Smithsonian Institution National Museum of Natural History MRC104 10th ST & Constitution Ave, NW Washington DC 20560-0104 202-357-1651
Please disregard my voice mail request for your equipment info and address - I didn't realize you had sent it to Mark already. I'm sorry you have felt bounced around, I hope that we have properly addressed (at least initially) your questions. I have some additional information that I will send to you as well. Ted Pella and Cressington are both very interested in you achieving success in your project. Please let me know what else I can do to support you.
Sincerely,
James
James Long Materials Science Specialist Ted Pella, Inc. 512-657-0898 james_long-at-tedpella.com www.tedpella.com
Mike - Topcon discontinued service support years ago. We have an ISI (Topcon) SEM that we continue to have serviced by our former ISI service engineer, who is independent now. His name is Chuck Humphrey and his company's name is Scanning Solutions. He has always done an excellent job for us. You can reach him the following ways: telephone 407 234 0676 fax 407 277 4423 email cchumph-at-attglobal.net
Good luck with the scope.
Sincerely,
Jill
Jill Verlander Reed, D.V.M. Associate Scientist Director,Electron Microscopy Core Facility UF College of Medicine P.O. Box 100215 HSC 1600 SW Archer Rd. Room RB-167 Gainesville, FL 32610-0215
-----Original Message----- } From: Mike Coviello [mailto:coviello-at-mae.uta.edu] Sent: Thursday, December 20, 2001 4:58 PM To: verlaj-at-mail-cs.med.ufl.edu
Hi All: I was asked to look at an old SEM we have and I am looking to see if there is company support for an old ISI SEM. I guess thay have become Topcon ? Do they support older ISI SEM's. Let me know if you have any knowledge. Thanks in advance. Thx, Mike Coviello UTA
As intended, detailed responses to many helpful topics are often generated by the group. However, searching through archives for the discussions that one vaguely remembers from several years ago is quite difficult given the limitation of a month by month search.
My thought was that perhaps a listing of "Best Of" discussions could be made that might be as simple as a listing of topics and the starting date of the discussion. Interested parties could then look through this summary before searching the monthly archives. The monthly archives would then be much more useful and repeats of common questions would be reduced. (A similar benefit could be achieved by providing a search capability of just the subject lines for each year.)
Overall, this listserver is a very valuable resource, and this suggestion may be a relatively easy way to make it even more useful.
Happy Holidays to All!
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
} was asked to look at an old SEM we have and I am looking } to see if there is company support for an old ISI SEM. I guess } thay have become Topcon ? Do they support older ISI SEM's. } Let me know if you have any knowledge. Thanks in advance.
It is my understanding that ISI became Topcon, which in the US became part of RJ Lee, which later became Aspex Instruments ("www.aspexllc.com" or "www.rjleeinst.com"). I have no idea if they support the ISI line.
You can find a list of third party SEM service providers at "www.jcnabity.com/service.htm".
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
I am a graduate student working on a bacterial ecology project, and attempting to find a vendor for a tunable dye laser to replace my mercury lamp in exciting the eAFP (GFP/CFP/YFP) that my bacteria produce. I would like to be able to move fairly rapidly between widely different frequencies, so that I can later false color a frame with relative ease.
I was looking at Laser Science (www.laserscience.com) and have been having major trouble getting timely service from them even in shipping a demo unit, replying to email/phone calls, etc.
I would like to know if there are more established or recommended vendors, and if anyone has experience with a machine, can give interesting data such as price range, etc.
My email is: bckirkup-at-post.harvard.edu; my apologies that even as an MSA member, I do not regularly read the listserv itself, so if you would please reply privately, thank you.
We are looking for an experienced person to do TEM including specimen preparation, embedding, sectioning, staining, and some microscopy on individual biomedical projects. A part time position with very flexible hours, ideal for someone who would like to work this into life. We are building a center for microscopy in children's diseases at Children's Hospital Oakland and have more and more projects from within the research institute and from collaborations with physicians and scientists at other institutions in Northern California. We would like one or more people to be able to carry out parts of the individual projects, working closely together with us. Someone who takes pride in doing superb work, who has the experience to help choose the most appropriate techniques for each project, and who doesn't mind travelling to our sister institutions to use particular instruments or help with experiments. I would be happy to correspond with anyone interested at {jbastacky-at-chori.org} . Please send a portfolio of your micrographs. Children's Hospital is an equal opportunity employer. -- Jacob Bastacky, M.D. Research Physician Children's Hospital Oakland Research Institute 5700 Martin Luther King Jr. Way Oakland, California 94609 Telephone: 510.450.7639 email: jbastacky-at-CHORI.org FAX: 510.450.7910
Senior Scientist, Visiting National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 sjbastacky-at-lbl.gov
This post was written in response to the question by Scott Whittaker regarding Ni sputtering in a Cressington 108 sputter coater. Disclaimer: It contains both material on Ni sputtering in general as well as specific information regarding the Cressington instrument line that Mr. Whittaker brings up in his question. Since Mr. Whittaker brought it up, we felt that it was important to respond with specific information regarding instrument models; otherwise, a general answer might still leave some possibility of confusion. This is not meant as an “ad”, and we hope it is not taken as such.
First, as mentioned by others, a planar magnetron system is certainly capable of sputtering Ni, and at a much lower cost compared to an ion beam system. You mention that you have a Cressington 108 system. After some offline correspondence we've determined that you have a 108autoSE system. This is important to know, since different Cressington systems have different Ni sputtering capabilities and other Cressington users who read this response should be clear about the capability of their particular unit.
Two major issues in sputtering Ni concern a coater’s vacuum system and magnet system. Regarding the vacuum system, pumping capability affects the ratio of argon to background undesirables (oxygen, water) and affects what happens chemically at the target. Materials that oxidize are going to do so much more readily in a 108 series unit. Your 108autoSE as well as the 108auto use a rotary pump, while a 208HR uses both a rotary and a molecular drag pump. In short, the 208HR is pumping the chamber more than 30 times faster than either 108. It is more capable of handling Ni, which quickly oxidizes.
Regarding the magnet system, the strength of the magnet combined with the thickness of the target will determine your sputtering capability. A weaker magnet necessitates a thinner target. Your 108auto/SE as well as a 208HR have a much more powerful magnet than a 108auto. Any attempt to sputter Ni in a 108auto with a thick target will fail as the magnetic field cannot penetrate to the active surface of the target. Using thin targets (.1mm) may work but the rates will be slow. Even though your 108auto/SE has a strong magnet system, sputtering should also only be attempted with thin targets to optimize your setup.
There are those who use the 108auto to produce Ni films for EDX but it is not the optimum route. The 108auto/SE would be better with its superior magnetic system. The best choice is the 208HR with good gas handling and magnetic systems. The 208HR sputters Ni readily with targets up to 2mm thick.
I have more detailed product information that I would be happy to share with anyone offline.
Disclaimer: Ted Pella, Inc. is now the exclusive representative of the Cressington EM products in the Americas and also sells sputter targets.
James Long Materials Science Specialist Ted Pella, Inc. 512-657-0898 james_long-at-tedpella.com www.tedpella.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cg27-at-wwa.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, December 21, 2001 at 13:33:29 ---------------------------------------------------------------------------
Email: cg27-at-wwa.com Name: Paul
Organization: none
Education: Graduate College
Location: DuPage County, IL
Question: Hello,
I am an adult hobbyist with a rather old but nice B&L microscope. Can you recommend any good reference books that would help me identify the various microbes, bacteria, etc. that I encounter here in Cook/DuPage? I work within the 100x - 500x range using very little preparation; my 1000x oil-imms optic is decent but rarely gets used due to its "high-maintenance" character.
Don't hesitate to recommend a children's book (that's all I've been able to find!) if it's particularly good. I'm looking for a good, quick reference that is well-illustrated; if it also contains theory - all the better!!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (david.m.heckman-at-pharmacia.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, December 26, 2001 at 12:55:37 ---------------------------------------------------------------------------
Email: david.m.heckman-at-pharmacia.com Name: David Heckman
Organization: Pharmacia
Education: Graduate College
Location: Kalamazoo, MI USA
Question: how do I use an "oil immersion" lense, and how do I determine which oil to use and what refraction rating to use.
I need two Bausch & Lomb Clear Glass Plates (Cat. No. 31-26-86) to replace missing plates in the base stands of two binocular dissecting scopes. Can someone please tell me where I might order these (from B&L or other supplier). I can only find internet sites to the Bausch & Lomb contact lens business. Thank you.
Dean Abel Biological Sciences 138BB University of Iowa Iowa City IA 52242 319-335-1070
I think the originals are available but quite expensive - to replace severaql I needed I went to a local glass shop and they cut them out of 1/4" plate and polished the edges - for about $8.00 each - B&L were about $24 when you could find them. You can also make them yourself out of Lexan or plexiglass - I have been able to do these myself out of scrap material and they worked fine and don't break. If you need more details on making the Lexan ones (very simple with a router - takes me about a minute each) get back to me.
Stephen Poe USDA, APHIS, PPQ Riverdale, MD
Dean Abel {dean-abel-at-uiowa.edu} on 12/27/2001 04:54:49 PM
To: Microscopy Society of America {Microscopy-at-sparc5.microscopy.com} cc:
I need two Bausch & Lomb Clear Glass Plates (Cat. No. 31-26-86) to replace missing plates in the base stands of two binocular dissecting scopes. Can someone please tell me where I might order these (from B&L or other supplier). I can only find internet sites to the Bausch & Lomb contact lens business. Thank you.
Dean Abel Biological Sciences 138BB University of Iowa Iowa City IA 52242 319-335-1070
Seasons Greetings and Happy New Year to Everyone !
The Call for Papers for the Microscopy and Microanalysis 2002 meeting is now on-line.
All the details for the meeting: pre-conference congresses, short courses, tutorials, symposia, awards, travel, exhibition, registration, lodging and paper submission are now avialable available at:
http://www.msa.microscopy.com/MMHomePage.html
Join MSA, MAS, MSC/SMC and IMS in Quebec City August 4-8, 2002.
You may also download a PDF copy of the complete Call for Papers from this WWW Site.
See you in August .
Nestor Your Friendly Neighborhood SysOp
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