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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
From daemon Wed Jan 31 20:11:04 2001
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